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WO2025179213A1 - Targeting trop-2 in diffuse pleural mesothelioma - Google Patents

Targeting trop-2 in diffuse pleural mesothelioma

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Publication number
WO2025179213A1
WO2025179213A1 PCT/US2025/016899 US2025016899W WO2025179213A1 WO 2025179213 A1 WO2025179213 A1 WO 2025179213A1 US 2025016899 W US2025016899 W US 2025016899W WO 2025179213 A1 WO2025179213 A1 WO 2025179213A1
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WIPO (PCT)
Prior art keywords
antibody
trop
seq
antibodies
expression
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
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PCT/US2025/016899
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French (fr)
Inventor
Michael OFFIN
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Memorial Sloan Kettering Cancer Center
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Memorial Sloan Kettering Cancer Center
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Publication of WO2025179213A1 publication Critical patent/WO2025179213A1/en
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/30—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6801—Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent
    • A61K47/6803—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
    • A61K47/68037—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates the drug being a camptothecin [CPT] or derivatives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6835—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site
    • A61K47/6851—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody targeting a determinant of a tumour cell
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6835—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site
    • A61K47/6851—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody targeting a determinant of a tumour cell
    • A61K47/6857—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody targeting a determinant of a tumour cell the tumour determinant being from lung cancer cell
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6889—Conjugates wherein the antibody being the modifying agent and wherein the linker, binder or spacer confers particular properties to the conjugates, e.g. peptidic enzyme-labile linkers or acid-labile linkers, providing for an acid-labile immuno conjugate wherein the drug may be released from its antibody conjugated part in an acidic, e.g. tumoural or environment
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
    • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
    • A61K51/04—Organic compounds
    • A61K51/08—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins
    • A61K51/10—Antibodies or immunoglobulins; Fragments thereof, the carrier being an antibody, an immunoglobulin or a fragment thereof, e.g. a camelised human single domain antibody or the Fc fragment of an antibody
    • A61K51/1045—Antibodies or immunoglobulins; Fragments thereof, the carrier being an antibody, an immunoglobulin or a fragment thereof, e.g. a camelised human single domain antibody or the Fc fragment of an antibody against animal or human tumor cells or tumor cell determinants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
    • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
    • A61K51/04—Organic compounds
    • A61K51/08—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins
    • A61K51/10—Antibodies or immunoglobulins; Fragments thereof, the carrier being an antibody, an immunoglobulin or a fragment thereof, e.g. a camelised human single domain antibody or the Fc fragment of an antibody
    • A61K51/1045—Antibodies or immunoglobulins; Fragments thereof, the carrier being an antibody, an immunoglobulin or a fragment thereof, e.g. a camelised human single domain antibody or the Fc fragment of an antibody against animal or human tumor cells or tumor cell determinants
    • A61K51/1054—Antibodies or immunoglobulins; Fragments thereof, the carrier being an antibody, an immunoglobulin or a fragment thereof, e.g. a camelised human single domain antibody or the Fc fragment of an antibody against animal or human tumor cells or tumor cell determinants the tumor cell being from lung
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
    • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
    • A61K51/04—Organic compounds
    • A61K51/08—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins
    • A61K51/10—Antibodies or immunoglobulins; Fragments thereof, the carrier being an antibody, an immunoglobulin or a fragment thereof, e.g. a camelised human single domain antibody or the Fc fragment of an antibody
    • A61K51/1093—Antibodies or immunoglobulins; Fragments thereof, the carrier being an antibody, an immunoglobulin or a fragment thereof, e.g. a camelised human single domain antibody or the Fc fragment of an antibody conjugates with carriers being antibodies
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the present disclosure provides a method for treating diffuse pleural mesothelioma (DPM) in a subject in need thereof comprising administering to the subject an effective amount of an antibody-drug conjugate comprising an anti-TROP-2 antibody or antigen binding fragment thereof.
  • the present disclosure provides a method for treating diffuse pleural mesothelioma (DPM) in a subject in need thereof comprising administering to the subject an effective amount of an antibody-drug conjugate comprising an anti-TROP-2 -1- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 antibody or antigen binding fragment thereof and an effective amount of an AKT inhibitor.
  • the present disclosure provides a method for re-sensitizing DPM tumors to anti-TROP-2 antibody-drug conjugate therapy comprising administering to the subject an effective amount of an antibody-drug conjugate comprising an anti-TROP-2 antibody or antigen binding fragment thereof and an effective amount of an AKT inhibitor.
  • the antibody-drug conjugate and the AKT inhibitor are administered separately, simultaneously, or sequentially.
  • AKT inhibitors include, but are not limited to, samotolisib, oridonin, capivasertib, ipatasertib, miransertib, afuresertib, uprosertib, BAY1125976, MK-2206, TAS-117, GSK690693, triciribine, and perifosine.
  • the AKT inhibitor is administered intravenously, intraperitoneally, subcutaneously, intramuscularly, or intratumorally.
  • the antibody-drug conjugate comprises one or more of alkylating agents, topoisomerase inhibitors, platinum agents, taxanes, vinca agents, anti-estrogen drugs, aromatase inhibitors, ovarian suppression agents, VEGF/VEGFR inhibitors, EGF/EGFR inhibitors, PARP inhibitors, cytostatic alkaloids, cytotoxic antibiotics, antimetabolites, endocrine/hormonal agents, and bisphosphonate therapy agents.
  • the antibody-drug conjugate comprises a cytotoxic drug selected from the group consisting of an anthracycline, a camptothecin, a tubulin inhibitor, a maytansinoid, a calicheamycin, an auristatin, a nitrogen mustard, an ethylenimine derivative, an alkyl sulfonate, a nitrosourea, a triazene, a folic acid analog, a taxane, a COX-2 inhibitor, a pyrimidine analog, a purine analog, an antibiotic, an enzyme inhibitor, an epipodophyllotoxin, a platinum coordination complex, a vinca alkaloid, a substituted urea, a methyl hydrazine derivative, an adrenocortical suppressant, a hormone antagonist, an antimetabolite, an alkylating agent, an antimitotic, an anti-angiogenic agent, a tyrosine
  • a cytotoxic drug selected from the
  • the antibody-drug conjugate comprises cyclophosphamide, fluorouracil (or 5- fluorouracil or 5-FU), methotrexate, edatrexate (10-ethyl-10-deaza-aminopterin), thiotepa, carboplatin, cisplatin, taxanes, paclitaxel, protein-bound paclitaxel, docetaxel, vinorelbine, tamoxifen, raloxifene, toremifene, fulvestrant, gemcitabine, irinotecan, ixabepilone, -2- 4932-3602-9469.1 Atty.
  • the anti- TROP-2 antibody or antigen binding fragment comprises the variable heavy (VH) domain and variable light (VL) domain of sacituzumab.
  • the V H domain comprises a V H -CDR1 sequence of SEQ ID NO: 5, a V H - CDR2 sequence of SEQ ID NO: 6, and a VH-CDR3 sequence of SEQ ID NO: 7
  • the VL domain comprises a VL-CDR1 sequence of SEQ ID NO: 8, a VL-CDR2 sequence of SEQ ID NO: 9, and a V L -CDR3 sequence of SEQ ID NO: 10.
  • the V H domain comprises an amino acid sequence of SEQ ID NO: 3; and (b) the VL domain comprises an amino acid sequence of SEQ ID NO: 1.
  • the antibody-drug conjugate comprises sacituzumab govitecan. -3- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 [0012]
  • the anti-TROP-2 antibody or antigen binding fragment comprises the variable heavy domain and variable light domain of datopotamab.
  • the VH domain comprises a VH-CDR1 sequence of SEQ ID NO: 15, a VH-CDR2 sequence of SEQ ID NO: 16, and a VH-CDR3 sequence of SEQ ID NO: 17 and the V L domain comprises a V L -CDR1 sequence of SEQ ID NO: 18, a V L -CDR2 sequence of SEQ ID NO: 19, and a V L -CDR3 sequence of SEQ ID NO: 20.
  • the VH domain comprises an amino acid sequence of SEQ ID NO: 13; and (b) the V L domain comprises an amino acid sequence of SEQ ID NO: 11.
  • the subject has received a prior anti-cancer therapy. Additionally or alternatively, in some embodiments, the subject is diagnosed with or is suffering from recurrent DPM or metastatic DPM. In any and all embodiments of the methods disclosed herein, the antibody-drug conjugate is administered intravenously, intraperitoneally, subcutaneously, intramuscularly, or intratumorally. BRIEF DESCRIPTION OF THE DRAWINGS [0014] FIGs.1A-1C. TROP-2 expression enriched in malignant pleural mesothelioma and associated with worse clinical outcomes.
  • TMA tissue microarray
  • To the left are representative TROP-2 immunohistochemistry in an example of benign pleura (1) with H-score of 0 and in DPM (2-4) with H-scores of 167, 24 and 11, respectively (original magnifications 200x).
  • FIG.1B Overall survival (OS) of patients whose samples were included in the DPM TMA noting significantly worse clinical outcomes in patients with TROP- 2 H-Score ⁇ 10.
  • FIG.1C Forrest plot evaluating risk of death using a cox regression analysis noting significantly improved outcomes in patients with low TROP-2 expression (defined as an H-score ⁇ 10) and in those with epithelioid histology.
  • FIGs.2A-2D TROP-2 expression is associated to the activation of pro- oncogenic pathways in DPM.
  • FIG.2A Dotplots depicting pathway enrichment analyses in two independent transcriptomic datasets of DPM clinical specimens (TCGA(4) and -4- 4932-3602-9469.1 Atty. Dkt.
  • FIG.2B Heatmaps showing expression of genes of interest, particularly upregulated genes involved in stemness, EMT/metastasis, and MAPK and AKT signaling in TCGA and Mesomics datasets.
  • FIG.2C Western blot showing dysregulation of protein markers of interest in isogenic DPM cell lines with ectopic TROP-2 expression dysregulation. Western blotting was reproduced in independent lysates (biological replicates), and a representative image is shown.
  • TROP-2 TROP-2 overexpression
  • sgTROP- 2 TROP-2 KO
  • FIG.2D Dotplots showing pathway enrichment analyses after DGE on isogenic TROP-2-overexpressing (TROP-2) or TROP-2-KO (sgTROP-2) cell lines compared to their control counterpart (Control).
  • FIGs.3A-3F TROP-2 exerts pro-oncogenic effects in DPM preclinical models.
  • FIG.3A Proliferation curves of isogenic TROP-2-overexpressing (TROP-2) or TROP-2-KO (sgTROP-2) cell lines compared to their control counterpart.
  • FIG.3B In vitro soft agar colony formation (FIG.3B), migration (FIG.3C) and invasion (FIG.3D) of isogenic TROP-2-overexpressing (TROP-2) or TROP-2-KO (sgTROP-2) cell lines compared to their control counterpart.
  • FIG.3F Survival curves of mice included in the intracardiac injection experiments in FIG.3E.
  • FIG.4A Heatmap showing TROP-2 expression at the mRNA level (transcriptomic sequencing, in logTPM (transcripts per million ) units) and at the protein level (immunohistochemistry, in H-score, and flow cytometry, in logMFI (mean fluorescence intensity) units).
  • FIG.4C Dotplots showing pathway enrichment analyses after DGE on SG-treated versus control MSK_Lx307 and MSK_Lx707 DPM PDX tumors.
  • FIG.4D Western blots showing activation of the PI3K/AKT pathway by assessing phosphorylation of AKT and its downstream effector PRAS40 in SG-treated versus control MSK_Lx307 and MSK_Lx707 DPM PDX tumors.
  • FIG.4E In vitro synergy assays testing the synergic potential of the combination of SG with the AKT/mTOR inhibitor samotolisib in DPM cell lines with TROP-2 expression, H2452 and TROP-2-ovexpressing MSTO-211H.
  • FIG.4H Corresponding ex vivo biodistribution of main organs Lx13 (blue) and Lx606 (red) at 72 hours and 120 hours post-injection of [89Zr]Zr-DFO- SG.
  • FIGS.5A-5D Median overall survival (OS) of patients with diffuse pleural mesotheliomas (DPM) based on TROP-2 expression in a tissue microarray (TMA) at various TROP-2 H-Score cut points: (FIG.5A) 5, (FIG.5B) 10, (FIG.5C) 16.5 [H-score cut point estimated to optimally divide the cohort by survival status], and (FIG.5D) 20.
  • FIG 6 IVIS imaging of immunocompromised mice right after injection of luciferase-expressing H2452 (control and TROP-2-KO) and H28 (control and TROP-2- overexpressing) to ensure equal administration of cells in all models.
  • FIGs.7A-7B FIG.7A: TROP-2 expression in DPM PDXs by IHC.
  • FIG.7B TROP-2 expression in 25 tested DPM POX models and 3 DPM cells lines by flow cytometry (PE anti-human TACSTD2 [TROP-2, clone NY18], Biolegend; lsotype control [blue]; TROP-2 [red marked with (*)]).
  • FIG.8B TROP-2 mRNA expression in MSK-Lx307 and MSK- Lx707 DPM PDXs, comparing SG-treated tumor versus the corresponding control tumor.
  • FIGs.9A-9C Radiochemical characterization and bioconjugation properties of [89Zr]Zr-DFO-SG utilizing instant thin layer chromatography (FIG.9A), serum stability of the radioimmunoconjugate through 144 hours (FIG.9B), and MALDI-TOF for determination of chelator degree of labeling (FIG.9C).
  • FIG.10 shows the tissue microarray patient demographics.
  • PCR 1 A Practical Approach (IRL Press at Oxford University Press); MacPherson et al. (1995) PCR 2: A Practical Approach; Harlow and Lane eds. (1999) Antibodies, A Laboratory Manual; Freshney (2005) Culture of Animal Cells: A Manual of Basic Technique, 5th edition; Gait ed. (1984) Oligonucleotide Synthesis; U.S. Patent No.4,683,195; Hames and Higgins eds. (1984) Nucleic Acid Hybridization; Anderson (1999) Nucleic Acid Hybridization; Hames and Higgins eds.
  • TROP-2 transmembrane protein product of TACSTD2
  • ADCs antibody-drug conjugates
  • TROP-2-targeting ADCs are effective in treating aggressive DPM, and that AKT inhibitors can be used to successfully sensitize resistant DPM tumors to TROP-2-targeting ADCs.
  • the “administration” of an agent or drug to a subject includes any route of introducing or delivering to a subject a compound to perform its intended function. Administration can be carried out by any suitable route, including but not limited to, orally, intranasally, intrathecally, parenterally (intravenously, intramuscularly, intraperitoneally, or subcutaneously), rectally, intrathecally, intraocularly, intradermally, transmucosally, iontophoretically, or topically. Administration includes self-administration and the administration by another.
  • antibody collectively refers to immunoglobulins or immunoglobulin-like molecules including by way of example and without limitation, IgA, IgD, IgE, IgG and IgM, combinations and fragments thereof, and similar molecules produced during an immune response in any vertebrate, for example, in mammals such as humans, goats, rabbits and mice, as well as non-mammalian species, such as shark immunoglobulins.
  • Antibodies and antibody fragments can be wholly or partially derived from mammals (e.g., humans, non-human primates, goats, guinea pigs, hamsters, horses, mice, rats, rabbits and sheep) or non-mammalian antibody producing animals (e.g., chickens, ducks, geese, snakes, and urodele amphibians).
  • mammals e.g., humans, non-human primates, goats, guinea pigs, hamsters, horses, mice, rats, rabbits and sheep
  • non-mammalian antibody producing animals e.g., chickens, ducks, geese, snakes, and urodele amphibians.
  • the antibodies and antibody fragments can be produced in animals or produced outside of animals, such as from yeast or phage (e.g., as a single antibody or antibody fragment or as part of an antibody library).
  • antibodies includes intact immunoglobulins and “antigen binding fragments” specifically bind to a molecule of interest (or a group of highly similar molecules of interest) to the substantial exclusion of binding to other molecules (for example, antibodies and antibody fragments that have a binding constant for the molecule of interest that is at least 10 3 M -1 greater, at least 10 4 M -1 greater or at least 10 5 M -1 greater than a binding constant for other molecules in a biological sample).
  • antibody also includes genetically engineered forms such as chimeric antibodies (for example, humanized murine antibodies), heteroconjugate antibodies (such as, bispecific antibodies).
  • Antibodies may comprise whole native antibodies, monoclonal antibodies, human antibodies, humanized antibodies, camelised antibodies, multispecific antibodies, bispecific antibodies, chimeric antibodies, Fab, Fab', single chain V region fragments (scFv), single domain antibodies (e.g., nanobodies and single domain camelid antibodies), VNAR fragments, Bi-specific T- cell engager (BiTE) antibodies, minibodies, disulfide-linked Fvs (sdFv), and anti-idiotypic -9- 4932-3602-9469.1 Atty.
  • antibody refers to a polypeptide ligand comprising at least a light chain immunoglobulin variable region or heavy chain immunoglobulin variable region which specifically recognizes and binds an epitope of an antigen.
  • Antibodies are composed of a heavy and a light chain each of which has a variable region, termed the variable heavy (V H ) region and the variable light (V L ) region.
  • the heavy chain constant (C H ) region is comprised of three domains, CH1, CH2, and CH3.
  • an immunoglobulin has heavy (H) chains and light (L) chains interconnected by disulfide bonds. There are two types of light chain, lambda ( ⁇ ) and kappa ( ⁇ ). There are five main heavy chain classes (or isotypes) which determine the functional activity of an antibody molecule: IgM, IgD, IgG, IgA and IgE.
  • Immunoglobulin molecules can be of any type (e.g., IgG, IgE, IgM, IgD, IgA, and IgY), class (e.g., IgGl, IgG2, IgG3, IgG4, IgAl, and IgA2), or subclass.
  • Each heavy and light chain contains a constant region and a variable region, (the regions are also known as “domains”).
  • the heavy and the light chain variable regions specifically bind the antigen.
  • Light and heavy chain variable regions contain a “framework” region interrupted by three hypervariable regions, also called “complementarity-determining regions” or “CDRs”.
  • Each V H and V L is composed of three CDRs and four FRs arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4.
  • the framework region of an antibody that is the combined framework regions of the constituent light and heavy chains, largely adopt a ⁇ -sheet conformation and the CDRs form loops which connect, and in some cases form part of, the ⁇ -sheet structure.
  • framework regions act to form a scaffold that provides for positioning the CDRs in correct orientation by inter-chain, non-covalent interactions.
  • the CDRs are primarily responsible for binding to an epitope of an antigen.
  • the CDRs of each chain are typically referred to as CDR1, CDR2, and CDR3 and are also typically identified by the chain in which the particular CDR is located.
  • a VH CDR3 -10- 4932-3602-9469.1 Atty. Dkt.
  • No.115872-3182 is located in the variable domain of the heavy chain of the antibody in which it is found, whereas a VL CDR1 is the CDR1 from the variable domain of the light chain of the antibody in which it is found.
  • An antibody that binds TROP-2 protein will have a specific VH region and VL region sequence, and thus specific CDR sequences.
  • Antibodies with different specificities i.e. different combining sites for different antigens
  • SDRs specificity determining residues
  • Immunoglobulin-related compositions refers to antibodies (including monoclonal antibodies, polyclonal antibodies, humanized antibodies, chimeric antibodies, recombinant antibodies, multi-specific antibodies, bispecific antibodies, etc.,) as well as antibody fragments. An antibody or antigen binding fragment thereof specifically binds to an antigen.
  • antibody-related polypeptide means antigen-binding antibody fragments, including single-chain antibodies, that can comprise the variable region(s) alone, or in combination, with all or part of the following polypeptide elements: hinge region, CH1, CH2, and CH3 domains of an antibody molecule.
  • variable region(s) and hinge region CH 1 , CH 2 , and CH 3 domains.
  • Antibody-related molecules useful in the present methods e.g., but are not limited to, Fab, Fab′ and F(ab′)2, Fd, single-chain Fvs (scFv), single-chain antibodies, disulfide-linked Fvs (sdFv) and fragments comprising either a V L or V H domain.
  • Examples include: (i) a Fab fragment, a monovalent fragment consisting of the V L , V H , C L and CH 1 domains; (ii) a F(ab′)2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; (iii) a Fd fragment consisting of the VH and CH1 domains; (iv) a Fv fragment consisting of the V L and V H domains of a single arm of an antibody, (v) a dAb fragment (Ward et al., Nature 341: 544-546, 1989), which consists of a VH domain; and (vi) an isolated complementarity determining region (CDR).
  • a Fab fragment a monovalent fragment consisting of the V L , V H , C L and CH 1 domains
  • a F(ab′)2 fragment a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region
  • antibody fragments or “antigen binding fragments” can comprise a portion of a full length antibody, generally the antigen binding or variable region thereof.
  • antibody fragments or antigen binding fragments include Fab, Fab', F(ab')2, and Fv fragments; diabodies; linear antibodies; single-chain antibody molecules; and multi-specific antibodies formed from antibody fragments. -11- 4932-3602-9469.1 Atty. Dkt.
  • diabodies refers to small antibody fragments with two antigen-binding sites, which fragments comprise a heavy-chain variable domain (VH) connected to a light-chain variable domain (V L ) in the same polypeptide chain (V H V L ).
  • VH heavy-chain variable domain
  • V L light-chain variable domain
  • V H V L light-chain variable domain
  • linker that is too short to allow pairing between the two domains on the same chain, the domains are forced to pair with the complementary domains of another chain and create two antigen binding sites.
  • Diabodies are described more fully in, e.g., EP 404,097; WO 93/11161; and Hollinger et al., Proc. Natl. Acad. Sci. USA, 90: 6444-6448 (1993).
  • single-chain antibodies or “single-chain Fv (scFv)” refer to an antibody fusion molecule of the two domains of the Fv fragment, V L and V H .
  • Single-chain antibody molecules may comprise a polymer with a number of individual molecules, for example, dimer, trimer or other polymers.
  • the two domains of the Fv fragment, VL and VH are coded for by separate genes, they can be joined, using recombinant methods, by a synthetic linker that enables them to be made as a single protein chain in which the VL and VH regions pair to form monovalent molecules (known as single-chain Fv (scFv)).
  • scFv single-chain Fv
  • antibody fragments are obtained using conventional techniques known to those of skill in the art, and the fragments are screened for binding specificity and neutralization activity in the same manner as are intact antibodies.
  • antigen binding fragment refers to a fragment of the whole immunoglobulin structure which possesses a part of a polypeptide responsible for binding to antigen.
  • antigen binding fragment examples include scFv, (scFv)2, scFvFc, Fab, Fab′ and F(ab′)2, but are not limited thereto. Any of the above- noted antibody fragments are obtained using conventional techniques known to those of skill in the art, and the fragments are screened for binding specificity and neutralization activity in the same manner as are intact antibodies.
  • an “antigen” refers to a molecule to which an antibody (or antigen binding fragment thereof) can selectively bind.
  • the target antigen may be a protein, carbohydrate, nucleic acid, lipid, hapten, or other naturally occurring or synthetic compound.
  • the target antigen may be a polypeptide (e.g., a TROP-2 -12- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 polypeptide).
  • An antigen may also be administered to an animal to generate an immune response in the animal.
  • binding affinity is meant the strength of the total noncovalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen or antigenic peptide).
  • the affinity of a molecule X for its partner Y can generally be represented by the dissociation constant (K D ). Affinity can be measured by standard methods known in the art, including those described herein.
  • biological sample means sample material derived from living cells.
  • Biological samples may include tissues, cells, protein or membrane extracts of cells, and biological fluids (e.g., ascites fluid or cerebrospinal fluid (CSF)) isolated from a subject, as well as tissues, cells and fluids present within a subject.
  • biological fluids e.g., ascites fluid or cerebrospinal fluid (CSF)
  • Biological samples of the present technology include, but are not limited to, samples taken from breast tissue, renal tissue, the uterine cervix, the endometrium, the head or neck, the gallbladder, parotid tissue, the prostate, the brain, the pituitary gland, kidney tissue, muscle, the esophagus, the stomach, the small intestine, the colon, the liver, the spleen, the pancreas, thyroid tissue, heart tissue, lung tissue, the bladder, adipose tissue, lymph node tissue, the uterus, ovarian tissue, adrenal tissue, testis tissue, the tonsils, thymus, blood, hair, buccal, skin, serum, plasma, CSF, semen, prostate fluid, seminal fluid, urine, feces, sweat, saliva, sputum, mucus, bone marrow, lymph, and tears.
  • Biological samples can also be obtained from biopsies of internal organs or from cancers. Biological samples can be obtained from subjects for diagnosis or research or can be obtained from non-diseased individuals, as controls or for basic research. Samples may be obtained by standard methods including, e.g., venous puncture and surgical biopsy. In certain embodiments, the biological sample is a tissue sample obtained by needle biopsy.
  • CDR-grafted antibody means an antibody in which at least one CDR of an “acceptor” antibody is replaced by a CDR “graft” from a “donor” antibody possessing a desirable antigen specificity.
  • chimeric antibody means an antibody in which the Fc constant region of a monoclonal antibody from one species (e.g., a mouse Fc constant -13- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 region) is replaced, using recombinant DNA techniques, with an Fc constant region from an antibody of another species (e.g., a human Fc constant region).
  • a monoclonal antibody from one species e.g., a mouse Fc constant -13- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 region
  • conjugated refers to the association of two molecules by any method known to those in the art. Suitable types of associations include chemical bonds and physical bonds. Chemical bonds include, for example, covalent bonds and coordinate bonds.
  • Physical bonds include, for instance, hydrogen bonds, dipolar interactions, van der Waal forces, electrostatic interactions, hydrophobic interactions and aromatic stacking.
  • Consensus FR means a framework (FR) antibody region in a consensus immunoglobulin sequence. The FR regions of an antibody do not contact the antigen.
  • a "control" is an alternative sample used in an experiment for comparison purpose.
  • a control can be "positive” or "negative.”
  • a positive control a compound or composition known to exhibit the desired therapeutic effect
  • a negative control a subject or a sample that does not receive the therapy or receives a placebo
  • the term “effective amount” refers to a quantity sufficient to achieve a desired therapeutic and/or prophylactic effect, e.g., an amount which results in the prevention of, or a decrease in a disease or condition described herein or one or more signs or symptoms associated with a disease or condition described herein.
  • the amount of a composition administered to the subject will vary depending on the composition, the degree, type, and severity of the disease -14- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 and on the characteristics of the individual, such as general health, age, sex, body weight and tolerance to drugs. The skilled artisan will be able to determine appropriate dosages depending on these and other factors.
  • the compositions can also be administered in combination with one or more additional therapeutic compounds.
  • the therapeutic compositions may be administered to a subject having one or more signs or symptoms of a disease or condition described herein.
  • a "therapeutically effective amount" of a composition refers to composition levels in which the physiological effects of a disease or condition are ameliorated or eliminated. A therapeutically effective amount can be given in one or more administrations.
  • epitopes means a protein determinant capable of specific binding to an antibody. Epitopes usually consist of chemically active surface groupings of molecules such as amino acids or sugar side chains and usually have specific three dimensional structural characteristics, as well as specific charge characteristics. Conformational and non-conformational epitopes are distinguished in that the binding to the former but not the latter is lost in the presence of denaturing solvents.
  • an “epitope” of the TROP-2 protein is a region of the protein to which the anti-TROP-2 antibodies of the present technology specifically bind.
  • the epitope is a conformational epitope or a non-conformational epitope.
  • a routine cross-blocking assay such as that described in Antibodies, A Laboratory Manual, Cold Spring Harbor Laboratory, Ed Harlow and David Lane (1988), can be performed. This assay can be used to determine if an anti- TROP-2 antibody binds the same site or epitope as an anti-TROP-2 antibody of the present technology.
  • epitope mapping can be performed by methods known in the art.
  • the antibody sequence can be mutagenized such as by alanine scanning, to identify contact residues.
  • peptides corresponding to different regions of TROP-2 protein can be used in competition assays with the test antibodies or with a test antibody and an antibody with a characterized or known epitope.
  • expression includes one or more of the following: transcription of the gene into precursor mRNA; splicing and other processing of the precursor mRNA to produce mature mRNA; mRNA stability; translation of the mature mRNA into protein (including codon usage and tRNA availability); and glycosylation and/or other modifications of the translation product, if required for proper expression and function.
  • the term “gene” means a segment of DNA that contains all the information for the regulated biosynthesis of an RNA product, including promoters, exons, introns, and other untranslated regions that control expression.
  • “Homology” or “identity” or “similarity” refers to sequence similarity between two peptides or between two nucleic acid molecules. Homology can be determined by comparing a position in each sequence which may be aligned for purposes of comparison. When a position in the compared sequence is occupied by the same base or amino acid, then the molecules are homologous at that position.
  • a degree of homology between sequences is a function of the number of matching or homologous positions shared by the sequences.
  • a polynucleotide or polynucleotide region (or a polypeptide or polypeptide region) has a certain percentage (for example, at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99%) of “sequence identity” to another sequence means that, when aligned, that percentage of bases (or amino acids) are the same in comparing the two sequences.
  • This alignment and the percent homology or sequence identity can be determined using software programs known in the art. In some embodiments, default parameters are used for alignment.
  • One alignment program is BLAST, using default parameters.
  • Biologically equivalent polynucleotides are those having the specified percent homology and encoding a polypeptide having the same or similar biological activity.
  • humanized forms of non-human (e.g., murine) antibodies are chimeric antibodies which contain minimal sequence derived from non-human immunoglobulin.
  • humanized antibodies are human immunoglobulins in which hypervariable region residues of the recipient are replaced by hypervariable region residues from a non-human species (donor antibody) such as mouse, rat, rabbit or nonhuman primate having the desired specificity, affinity, and capacity.
  • donor antibody such as mouse, rat, rabbit or nonhuman primate having the desired specificity, affinity, and capacity.
  • Fv framework region (FR) residues of the human immunoglobulin are replaced by corresponding non-human residues.
  • humanized antibodies may -16- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 comprise residues which are not found in the recipient antibody or in the donor antibody. These modifications are made to further refine antibody performance such as binding affinity.
  • the humanized antibody will comprise substantially all of at least one, and typically two, variable domains (e.g., Fab, Fab′, F(ab′)2, or Fv), in which all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin and all or substantially all of the FR regions are those of a human immunoglobulin consensus FR sequence although the FR regions may include one or more amino acid substitutions that improve binding affinity.
  • variable domains e.g., Fab, Fab′, F(ab′)2, or Fv
  • the number of these amino acid substitutions in the FR are typically no more than 6 in the H chain, and in the L chain, no more than 3.
  • the humanized antibody optionally may also comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin.
  • Fc immunoglobulin constant region
  • hypervariable region refers to the amino acid residues of an antibody which are responsible for antigen-binding.
  • the hypervariable region generally comprises amino acid residues from a “complementarity determining region” or “CDR” (e.g., around about residues 24-34 (L1), 50-56 (L2) and 89-97 (L3) in the VL, and around about 31-35B (H1), 50-65 (H2) and 95-102 (H3) in the V H (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD.
  • CDR complementarity determining region
  • residues from a “hypervariable loop” e.g., residues 26-32 (L1), 50-52 (L2) and 91-96 (L3) in the V L , and 26-32 (H1), 52A-55 (H2) and 96-101 (H3) in the V H (Chothia and Lesk J. Mol. Biol.196:901-917 (1987)).
  • nucleic acids or polypeptide sequences refer to two or more sequences or subsequences that are the same or have a specified percentage of amino acid residues or nucleotides that are the same (i.e., about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or higher identity over a specified region (e.g., nucleotide sequence encoding an antibody described herein or amino acid sequence of an antibody described herein)), when compared and aligned for maximum correspondence over a comparison window or designated region as measured using a BLAST or BLAST 2.0 sequence comparison algorithms with default parameters described below, or by manual alignment and visual inspection (e.g., NCBI web site).
  • a specified region e.g., nucleotide sequence encoding an antibody described herein or amino acid sequence of an antibody described herein
  • sequences -17- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 are then said to be “substantially identical.”
  • This term also refers to, or can be applied to, the complement of a test sequence.
  • the term also includes sequences that have deletions and/or additions, as well as those that have substitutions.
  • identity exists over a region that is at least about 25 amino acids or nucleotides in length, or 50-100 amino acids or nucleotides in length.
  • the term “intact antibody” or “intact immunoglobulin” means an antibody that has at least two heavy (H) chain polypeptides and two light (L) chain polypeptides interconnected by disulfide bonds.
  • Each heavy chain is comprised of a heavy chain variable region (abbreviated herein as HCVR or V H ) and a heavy chain constant region.
  • the heavy chain constant region is comprised of three domains, CH 1 , CH 2 and CH 3 .
  • Each light chain is comprised of a light chain variable region (abbreviated herein as LCVR or VL) and a light chain constant region.
  • the light chain constant region is comprised of one domain, C L .
  • V H and V L regions can be further subdivided into regions of hypervariability, termed complementarity determining regions (CDR), interspersed with regions that are more conserved, termed framework regions (FR).
  • CDR complementarity determining regions
  • FR framework regions
  • Each VH and VL is composed of three CDRs and four FRs, arranged from amino-terminus to carboxyl-terminus in the following order: FR 1 , CDR 1 , FR 2 , CDR 2 , FR 3 , CDR 3 , FR 4 .
  • the variable regions of the heavy and light chains contain a binding domain that interacts with an antigen.
  • the constant regions of the antibodies can mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (Clq) of the classical complement system.
  • the term “monoclonal antibody” as used herein refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations that may be present in minor amounts.
  • a monoclonal antibody can be an antibody that is derived from a single clone, including any eukaryotic, prokaryotic, or phage clone, and not the method by which it is produced.
  • a monoclonal antibody composition displays a single binding specificity and affinity for a particular epitope.
  • Monoclonal antibodies are highly specific, being directed against a single antigenic site.
  • each monoclonal antibody is directed against a single determinant on the antigen.
  • the modifier “monoclonal” indicates the character of the antibody as being obtained from a substantially homogeneous -18- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 population of antibodies, and is not to be construed as requiring production of the antibody by any particular method.
  • Monoclonal antibodies can be prepared using a wide variety of techniques known in the art including, e.g., but not limited to, hybridoma, recombinant, and phage display technologies.
  • the monoclonal antibodies to be used in accordance with the present methods may be made by the hybridoma method first described by Kohler et al., Nature 256:495 (1975), or may be made by recombinant DNA methods (See, e.g., U.S. Patent No.4,816,567).
  • the “monoclonal antibodies” may also be isolated from phage antibody libraries using the techniques described in Clackson et al., Nature 352:624-628 (1991) and Marks et al., J. Mol.
  • the term “pharmaceutically-acceptable carrier” is intended to include any and all solvents, dispersion media, coatings, antibacterial and antifungal compounds, isotonic and absorption delaying compounds, and the like, compatible with pharmaceutical administration.
  • Pharmaceutically-acceptable carriers and their formulations are known to one skilled in the art and are described, for example, in Remington's Pharmaceutical Sciences (20 th edition, ed. A. Gennaro, 2000, Lippincott, Williams & Wilkins, Philadelphia, Pa.).
  • the term “polyclonal antibody” means a preparation of antibodies derived from at least two (2) different antibody-producing cell lines.
  • polynucleotide or “nucleic acid” means any RNA or DNA, which may be unmodified or modified RNA or DNA.
  • Polynucleotides include, without limitation, single- and double-stranded DNA, DNA that is a mixture of single- and double-stranded regions, single- and double-stranded RNA, RNA that is mixture of single- and double-stranded regions, and hybrid molecules comprising DNA and RNA that may be single-stranded or, more typically, double-stranded or a mixture of single- and double- stranded regions.
  • polynucleotide refers to triple-stranded regions comprising RNA or DNA or both RNA and DNA.
  • the term polynucleotide also includes DNAs or RNAs containing one or more modified bases and DNAs or RNAs with backbones modified for stability or for other reasons.
  • polypeptide As used herein, the terms “polypeptide,” “peptide” and “protein” are used interchangeably herein to mean a polymer comprising two or more amino acids joined to -19- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 each other by peptide bonds or modified peptide bonds, i.e., peptide isosteres.
  • Polypeptide refers to both short chains, commonly referred to as peptides, glycopeptides or oligomers, and to longer chains, generally referred to as proteins. Polypeptides may contain amino acids other than the 20 gene-encoded amino acids. Polypeptides include amino acid sequences modified either by natural processes, such as post-translational processing, or by chemical modification techniques that are well known in the art. Such modifications are well described in basic texts and in more detailed monographs, as well as in a voluminous research literature.
  • prevention or “preventing” of a disorder or condition refers to a compound that, in a statistical sample, reduces the occurrence of the disorder or condition in the treated sample relative to an untreated control sample, or delays the onset of one or more symptoms of the disorder or condition relative to the untreated control sample.
  • the term “recombinant” when used with reference, e.g., to a cell, or nucleic acid, protein, or vector indicates that the cell, nucleic acid, protein or vector, has been modified by the introduction of a heterologous nucleic acid or protein or the alteration of a native nucleic acid or protein, or that the material is derived from a cell so modified.
  • recombinant cells express genes that are not found within the native (non-recombinant) form of the cell or express native genes that are otherwise abnormally expressed, under expressed or not expressed at all.
  • the term “separate” therapeutic use refers to an administration of at least two active ingredients at the same time or at substantially the same time by different routes.
  • the term “sequential” therapeutic use refers to administration of at least two active ingredients at different times, the administration route being identical or different. More particularly, sequential use refers to the whole administration of one of the active ingredients before administration of the other or others commences.
  • “specifically binds” refers to a molecule (e.g., an antibody or antigen binding fragment thereof) which recognizes and binds another molecule (e.g., an antigen), but that does not substantially recognize and bind other molecules.
  • a molecule e.g., an antibody or antigen binding fragment thereof
  • Another molecule e.g., an antigen
  • No.115872-3182 “specific binding,” “specifically binds to,” or is “specific for” a particular molecule (e.g., a polypeptide, or an epitope on a polypeptide), as used herein, can be exhibited, for example, by a molecule having a K D for the molecule to which it binds to of about 10 ⁇ 4 M, 10 ⁇ 5 M, 10 ⁇ 6 M, 10 ⁇ 7 M, 10 ⁇ 8 M, 10 ⁇ 9 M, 10 ⁇ 10 M, 10 ⁇ 11 M, or 10 ⁇ 12 M.
  • binds may also refer to binding where a molecule (e.g., an antibody or antigen binding fragment thereof) binds to a particular polypeptide (e.g., a TROP-2 polypeptide), or an epitope on a particular polypeptide, without substantially binding to any other polypeptide, or polypeptide epitope.
  • a molecule e.g., an antibody or antigen binding fragment thereof
  • a particular polypeptide e.g., a TROP-2 polypeptide
  • epitope on a particular polypeptide without substantially binding to any other polypeptide, or polypeptide epitope.
  • the term “simultaneous” therapeutic use refers to the administration of at least two active ingredients by the same route and at the same time or at substantially the same time.
  • the terms “subject”, “patient”, or “individual” can be an individual organism, a vertebrate, a mammal, or a human.
  • the subject, patient, or individual is a human.
  • the term “therapeutic agent” is intended to mean a compound that, when present in an effective amount, produces a desired therapeutic effect on a subject in need thereof.
  • “Treating” or “treatment” as used herein covers the treatment of a disease or disorder described herein, in a subject, such as a human, and includes: (i) inhibiting a disease or disorder, i.e., arresting its development; (ii) relieving a disease or disorder, i.e., causing regression of the disorder; (iii) slowing progression of the disorder; and/or (iv) inhibiting, relieving, or slowing progression of one or more symptoms of the disease or disorder.
  • Therapeutic effects of treatment include, without limitation, inhibiting recurrence of disease, alleviation of symptoms, diminishment of any direct or indirect pathological consequences of the disease, preventing metastases, decreasing the rate of disease progression, amelioration or palliation of the disease state, and remission or improved prognosis.
  • treating a cancer is meant that the symptoms associated with the cancer are, e.g., alleviated, reduced, cured, or placed in a state of remission.
  • the various modes of treatment of disorders as described herein are intended to mean “substantial,” which includes total but also less than total treatment, and wherein some biologically or medically relevant result is achieved.
  • Dkt. No.115872-3182 treatment may be a continuous prolonged treatment for a chronic disease or a single, or few time administrations for the treatment of an acute condition.
  • Immunoglobulin-related Compositions of the Present Technology describes methods and compositions for the generation and use of anti-TROP-2 immunoglobulin-related compositions (e.g., anti-TROP-2 antibodies or antigen binding fragments thereof).
  • the anti-TROP-2 immunoglobulin- related compositions of the present disclosure may be useful in the treatment of anti-cancer drug-related skin rashes.
  • Anti-TROP-2 immunoglobulin-related compositions within the scope of the present technology include, e.g., but are not limited to, monoclonal, chimeric, humanized, bispecific antibodies and diabodies that specifically bind the target polypeptide, a homolog, derivative or a fragment thereof.
  • the present disclosure also provides antigen binding fragments of any of the anti-TROP-2 antibodies disclosed herein, wherein the antigen binding fragment is selected from the group consisting of Fab, F(ab)'2, Fab’, scF v , and F v .
  • the anti-TROP2 can comprise a commercially available antibody or antibody having the six CDRs of a commercially available antibody selected from the group consisting of LS-C126418, LS-C178765, LS-C126416, LS-C126417 (LifeSpan BioSciences, Inc., Seattle, Wash.); 10428-MM01, 10428-MM02, 10428-R001, 10428- R030 (Sino Biological Inc., Beijing, China); MR54 (eBioscience, San Diego, Calif.); sc- 376181, sc-376746, Santa Cruz Biotechnology (Santa Cruz, Calif.); MM0588- 49D6, (Novus Biologicals, Littleton, Colo.); ab79976, and ab89928 (ABCAM®, Cambridge, Mass.).
  • the anti-Trop-2 antibody can be selected from sacituzumab or another known anti-Trop antibody such as any of the following:
  • U.S. Publ. No. 2013/0089872 discloses anti-TROP2 antibodies K5-70 (Accession No. FERM BP- 11251), K5-107 (Accession No. FERM BP- 11252), K5-116-2-1 (Accession No. FERM BP-11253), T6-16 (Accession No. FERM BP- 11346), and T5-86 (Accession No. FERM BP-11254), deposited with the International Patent Organism Depositary, Tsukuba, Japan.
  • U.S. Pat. No.7,420,040 disclosed an anti-TROP2 antibody produced by hybridoma cell line AR47A6.4.2, deposited with the ID AC (International Depository Authority of Canada, Winnipeg, Canada) as accession number 141205-05.
  • U.S. Pat. No.7,420,041 disclosed an anti-TROP2 antibody produced by hybridoma cell line AR52A301.5, deposited with the -22- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 IDAC as accession number 141205-03.
  • No.2013/0122020 disclosed anti-Trop-2 antibodies 3E9, 6G11, 7E6, 15E2, 18B1. Hybridomas encoding a representative antibody were deposited with the American Type Culture Collection (ATCC), Accession Nos. PTA- 12871 and PTA-12872.
  • U.S. Pat. No.8,715,662 discloses anti-Trop-2 antibodies produced by hybridomas deposited at the AID-ICLC (Genoa, Italy) with deposit numbers PD 08019, PD 08020 and PD 08021.
  • U.S. Patent Application Publ. No.20120237518 discloses anti- TROP 2 antibodies 77220, KM4097 and KM4590.
  • the anti-TROP-2 antibody is sacituzumab, which is also known as the humanized monoclonal antibody hRS7 (e.g., U.S. Pat. No.7,238,785, incorporated herein by reference in its entirety).
  • the sacituzumab antibody was generated using a murine IgGl raised against a crude membrane preparation of a human primary squamous cell lung carcinoma. (Stein et al., Cancer Res.50: 1330, 1990).
  • the present disclosure provides a TROP-2 antibody or antigen binding fragment thereof comprising a heavy chain immunoglobulin variable domain (V H ) and a light chain immunoglobulin variable domain (VL), wherein the VH comprises a VH- CDR1 sequence of SEQ ID NO: 5, a VH-CDR2 sequence of SEQ ID NO: 6, and a VH- CDR3 sequence of SEQ ID NO:
  • the present disclosure provides a TROP-2 antibody or antigen binding fragment thereof comprising a heavy chain immunoglobulin variable domain (V H ) and a light chain immunoglobulin variable domain (V L ), wherein: (a) the V H domain comprises an amino acid sequence of SEQ ID NO: 3; and (b) the VL domain comprises an amino acid sequence of SEQ ID NO: 1.
  • the present disclosure provides a TROP-2 antibody or antigen binding fragment thereof comprising a heavy chain immunoglobulin variable domain (VH) and a light chain immunoglobulin variable domain (VL), wherein the VH comprises a VH- CDR1 sequence of SEQ ID NO: 15, a V H -CDR2 sequence of SEQ ID NO: 16, and a V H - CDR3 sequence of SEQ ID NO: 17 and the VL comprises a VL-CDR1 sequence of SEQ ID -26- 4932-3602-9469.1 Atty. Dkt.
  • VH heavy chain immunoglobulin variable domain
  • VL light chain immunoglobulin variable domain
  • the antibody further comprises a Fc domain of any isotype, e.g., but are not limited to, IgG (including IgG1, IgG2, IgG3, and IgG4), IgA (including IgA 1 and IgA 2 ), IgD, IgE, or IgM, and IgY.
  • IgG including IgG1, IgG2, IgG3, and IgG4
  • IgA including IgA 1 and IgA 2
  • IgD IgE
  • IgM IgM
  • Non-limiting examples of constant region sequences include: [0082] Human IgD constant region, Uniprot: P01880 (SEQ ID NO: 21) APTKAPDVFPIISGCRHPKDNSPVVLACLITGYHPTSVTVTWYMGTQSQPQRTFPEIQRRDS YYMTSSQLSTPLQQWRQGEYKCVVQHTASKSKKEIFRWPESPKAQASSVPTAQPQAEGSL AKATTAPATTRNTGRGGEEKKKEKEKEEQEERETKTPECPSHTQPLGVYLLTPAVQDLWLR DKATFTCFVVGSDLKDAHLTWEVAGKVPTGGVEEGLLERHSNGSQSQHSRLTLPRSLWNA GTSVTCTLNHPSLPPQRLMALREPAAQAPVKLSLNLLASSDPPEAASWLLCEVSGFSPPNILL MWLEDQREVNTSGFAPARPPPQPGSTTFWAWSVLRVPAPPSPQPATYTCVVSHEDSRTLLN ASRSLE
  • the immunoglobulin-related compositions of the present technology comprise a light chain constant region that is at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or is 100% identical to SEQ ID NO: 29.
  • the anti-TROP-2 immunoglobulin-related compositions of the present technology bind to an epitope within the extracellular region of human TROP-2.
  • the epitope is a conformational epitope or non-conformational epitope.
  • the TROP-2 polypeptide has the amino acid sequence of SEQ ID NO: 30.
  • the HC and LC immunoglobulin variable domain sequences are components of the same polypeptide chain. In other embodiments, the HC and LC immunoglobulin variable domain sequences are components of different polypeptide chains. In certain embodiments, the antibody is a full-length antibody. [0096] In some embodiments, the immunoglobulin-related compositions of the present technology bind specifically to at least one TROP-2 polypeptide.
  • the immunoglobulin-related compositions of the present technology bind at least one TROP-2 polypeptide with a dissociation constant (K D ) of about 10 ⁇ 3 M, 10 ⁇ 4 M, 10 ⁇ 5 M, 10 ⁇ 6 M, 10 ⁇ 7 M, 10 ⁇ 8 M, 10 ⁇ 9 M, 10 ⁇ 10 M, 10 ⁇ 11 M, or 10 ⁇ 12 M.
  • K D dissociation constant
  • the immunoglobulin-related compositions are monoclonal antibodies, chimeric antibodies, humanized antibodies, bispecific antibodies, or multi-specific antibodies.
  • the antibodies comprise a human antibody framework region.
  • the immunoglobulin-related composition includes one or more of the following characteristics: (a) a light chain immunoglobulin variable domain sequence that is at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the light chain immunoglobulin variable domain sequence of any one of SEQ ID NOs: 1 or 11; and/or (b) a heavy chain immunoglobulin variable domain sequence that is at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the heavy chain immunoglobulin variable domain sequence of any one of SEQ ID NOs: 3 or 13.
  • one or more amino acid residues in the immunoglobulin-related compositions provided herein are substituted with another amino acid.
  • the present disclosure provides an antibody comprising (a) a LC sequence that is at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the LC sequence present in SEQ ID NO: 2, or SEQ ID NO: 12; and/or (b) a HC sequence that is at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the HC sequence present in SEQ ID NO: 4, or SEQ ID NO: 14.
  • the immunoglobulin-related compositions contain an IgG1 constant region comprising one or more amino acid substitutions selected from the group consisting of N297A and K322A. Additionally or alternatively, in some -30- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 embodiments, the immunoglobulin-related compositions contain an IgG4 constant region comprising a S228P mutation. [00100] In some aspects, the anti-TROP-2 immunoglobulin-related compositions described herein contain structural modifications to facilitate rapid binding and cell uptake and/or slow release.
  • the anti-TROP-2 immunoglobulin-related composition of the present technology may contain a deletion in the CH2 constant heavy chain region to facilitate rapid binding and cell uptake and/or slow release.
  • a Fab fragment is used to facilitate rapid binding and cell uptake and/or slow release.
  • a F(ab)' 2 fragment is used to facilitate rapid binding and cell uptake and/or slow release.
  • the present technology provides a nucleic acid sequence encoding any of the immunoglobulin-related compositions described herein. Also disclosed herein are recombinant nucleic acid sequences encoding any of the antibodies described herein.
  • the present technology provides a host cell expressing any nucleic acid sequence encoding any of the immunoglobulin-related compositions described herein.
  • the immunoglobulin-related compositions of the present technology e.g., an anti-TROP-2 antibody
  • the immunoglobulin-related compositions of the present technology can be monospecific, bispecific, trispecific or of greater multi- specificity.
  • Multi-specific antibodies can be specific for different epitopes of one or more TROP-2 polypeptides or can be specific for both the TROP-2 polypeptide(s) as well as for heterologous compositions, such as a heterologous polypeptide or solid support material.
  • the immunoglobulin-related compositions of the present technology can further be recombinantly fused to a heterologous polypeptide at the N- or C-terminus or chemically conjugated (including covalently and non-covalently conjugations) to polypeptides or other compositions.
  • the immunoglobulin-related compositions of the present technology can be recombinantly fused or conjugated to molecules useful as labels in detection assays and effector molecules such as heterologous polypeptides, drugs, or toxins. -31- 4932-3602-9469.1 Atty. Dkt.
  • a functional group on the immunoglobulin-related composition typically associates with a functional group on the agent.
  • a functional group on the agent associates with a functional group on the immunoglobulin-related composition.
  • the functional groups on the agent and immunoglobulin-related composition can associate directly.
  • a functional group e.g., a sulfhydryl group
  • an agent can associate with a functional group (e.g., sulfhydryl group) on an immunoglobulin-related composition to form a disulfide.
  • the functional groups can associate through a cross-linking agent (i.e., linker).
  • the cross-linker can be attached to either the agent or the immunoglobulin-related composition.
  • the number of agents or immunoglobulin-related compositions in a conjugate is also limited by the number of functional groups present on the other.
  • the maximum number of agents associated with a conjugate depends on the number of functional groups present on the immunoglobulin-related composition.
  • the maximum number of immunoglobulin-related compositions associated with an agent depends on the number of functional groups present on the agent.
  • the conjugate comprises one immunoglobulin- related composition associated to one agent.
  • a conjugate comprises at least one agent chemically bonded (e.g., conjugated) to at least one immunoglobulin-related composition.
  • the agent can be chemically bonded to an immunoglobulin-related composition by any method known to those in the art.
  • a functional group on the agent may be directly attached to a functional group on the immunoglobulin-related composition.
  • suitable functional groups include, for example, amino, carboxyl, sulfhydryl, maleimide, isocyanate, isothiocyanate and hydroxyl. -32- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 [00108]
  • the agent may also be chemically bonded to the immunoglobulin-related composition by means of cross-linking agents, such as dialdehydes, carbodiimides, dimaleimides, and the like.
  • Cross-linking agents can, for example, be obtained from Pierce Biotechnology, Inc., Rockford, Ill. The Pierce Biotechnology, Inc. web-site can provide assistance. Additional cross-linking agents include the platinum cross-linking agents described in U.S. Pat. Nos.5,580,990; 5,985,566; and 6,133,038 of Kreatech Biotechnology, B.V., Amsterdam, The Netherlands. [00109] Alternatively, the functional group on the agent and immunoglobulin-related composition can be the same. Homobifunctional cross-linkers are typically used to cross- link identical functional groups.
  • homobifunctional cross-linkers examples include EGS (i.e., ethylene glycol bis[succinimidylsuccinate]), DSS (i.e., disuccinimidyl suberate), DMA (i.e., dimethyl adipimidate.2HCl), DTSSP (i.e., 3,3'- dithiobis[sulfosuccinimidylpropionate])), DPDPB (i.e., 1,4-di-[3'-(2'-pyridyldithio)- propionamido]butane), and BMH (i.e., bis-maleimidohexane).
  • EGS i.e., ethylene glycol bis[succinimidylsuccinate]
  • DSS i.e., disuccinimidyl suberate
  • DMA i.e., dimethyl adipimidate.2HCl
  • DTSSP i.e., 3,
  • Such homobifunctional cross-linkers are also available from Pierce Biotechnology, Inc. [00110] In other instances, it may be beneficial to cleave the agent from the immunoglobulin-related composition.
  • the web-site of Pierce Biotechnology, Inc. described above can also provide assistance to one skilled in the art in choosing suitable cross-linkers which can be cleaved by, for example, enzymes in the cell. Thus the agent can be separated from the immunoglobulin-related composition.
  • cleavable linkers examples include SMPT (i.e., 4-succinimidyloxycarbonyl-methyl-a-[2-pyridyldithio]toluene), Sulfo-LC- SPDP (i.e., sulfosuccinimidyl 6-(3-[2-pyridyldithio]-propionamido)hexanoate), LC-SPDP (i.e., succinimidyl 6-(3-[2-pyridyldithio]-propionamido)hexanoate), Sulfo-LC-SPDP (i.e., sulfosuccinimidyl 6-(3-[2-pyridyldithio]-propionamido)hexanoate), SPDP (i.e., N- succinimidyl 3-[2-pyridyldithio]-propionamidohexanoate), and AEDP
  • a conjugate comprises at least one agent physically bonded with at least one immunoglobulin-related composition.
  • Any method known to those in the art can be employed to physically bond the agents with the immunoglobulin-related compositions.
  • the immunoglobulin-related compositions and agents can be mixed together by any method known to those in the art. The order of mixing is not important.
  • agents can be physically mixed with immunoglobulin-related compositions by any method known to those in the art.
  • Target polypeptides within the scope of the present technology include any polypeptide derived from TROP-2 protein containing the extracellular domain which is capable of eliciting an immune response.
  • TROP-2 protein containing the extracellular domain which is capable of eliciting an immune response.
  • recombinantly engineered antibodies and antibody fragments e.g., antibody-related polypeptides, which are directed to TROP-2 protein and fragments thereof are suitable for use in accordance with the present disclosure.
  • Anti-TROP-2 antibodies that can be subjected to the techniques set forth herein include monoclonal and polyclonal antibodies, and antibody fragments such as Fab, Fab′, F(ab′) 2 , Fd, scFv, diabodies, antibody light chains, antibody heavy chains and/or antibody fragments. Methods useful for the high yield production of antibody Fv-containing polypeptides, e.g., Fab′ and F(ab′) 2 antibody fragments have been described. See U.S. Pat. No.5,648,237. [00116] Generally, an antibody is obtained from an originating species.
  • nucleic acid or amino acid sequence of the variable portion of the light chain, heavy chain or both, of an originating species antibody having specificity for a target polypeptide antigen is obtained.
  • An originating species is any species which was useful to generate the antibody of the present technology or library of antibodies, e.g., rat, mouse, rabbit, chicken, monkey, human, and the like.
  • Phage or phagemid display technologies are useful techniques to derive the antibodies of the present technology. Techniques for generating and cloning monoclonal antibodies are well known to those skilled in the art. Expression of sequences encoding antibodies of the present technology, can be carried out in E. coli.
  • nucleic acid coding sequences which encode substantially the same amino acid sequences as those of the naturally occurring proteins may be used in the practice of the present technology
  • sequences which encode substantially the same amino acid sequences as those of the naturally occurring proteins may be used in the practice of the present technology
  • These include, but are not limited to, nucleic acid -34- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 sequences including all or portions of the nucleic acid sequences encoding the above polypeptides, which are altered by the substitution of different codons that encode a functionally equivalent amino acid residue within the sequence, thus producing a silent change.
  • nucleotide sequence of an immunoglobulin tolerates sequence homology variations of up to 25% as calculated by standard methods (“Current Methods in Sequence Comparison and Analysis,” Macromolecule Sequencing and Synthesis, Selected Methods and Applications, pp.127-149, 1998, Alan R. Liss, Inc.) so long as such a variant forms an operative antibody which recognizes TROP-2 proteins.
  • one or more amino acid residues within a polypeptide sequence can be substituted by another amino acid of a similar polarity which acts as a functional equivalent, resulting in a silent alteration.
  • Substitutes for an amino acid within the sequence may be selected from other members of the class to which the amino acid belongs.
  • the nonpolar (hydrophobic) amino acids include alanine, leucine, isoleucine, valine, proline, phenylalanine, tryptophan and methionine.
  • the polar neutral amino acids include glycine, serine, threonine, cysteine, tyrosine, asparagine, and glutamine.
  • the positively charged (basic) amino acids include arginine, lysine and histidine.
  • the negatively charged (acidic) amino acids include aspartic acid and glutamic acid.
  • proteins or fragments or derivatives thereof which are differentially modified during or after translation, e.g., by glycosylation, proteolytic cleavage, linkage to an antibody molecule or other cellular ligands, etc.
  • an immunoglobulin encoding nucleic acid sequence can be mutated in vitro or in vivo to create and/or destroy translation, initiation, and/or termination sequences or to create variations in coding regions and/or form new restriction endonuclease sites or destroy pre-existing ones, to facilitate further in vitro modification. Any technique for mutagenesis known in the art can be used, including but not limited to in vitro site directed mutagenesis, J. Biol.
  • the antibody is an anti-TROP-2 monoclonal antibody.
  • the anti-TROP-2 monoclonal antibody may be a human or a mouse anti-TROP-2 monoclonal antibody.
  • any technique that provides for the production of antibody molecules by continuous cell line culture can be utilized. Such techniques include, but are not limited to, the hybridoma technique (See, e.g., Kohler & Milstein, 1975.
  • PCR utilizing primers derived from sequences encoding conserved regions of antibodies is used to amplify sequences encoding portions of antibodies from the population and then DNAs encoding antibodies or fragments thereof, such as variable domains, are reconstructed from the amplified sequences.
  • Such amplified sequences also can be fused to DNAs encoding other proteins ⁇ e.g., a bacteriophage coat, or a bacterial cell surface protein ⁇ for expression and display of the fusion polypeptides on phage or bacteria. Amplified sequences can then be expressed and further selected or isolated based, e.g., on the affinity of the expressed antibody or fragment thereof for an antigen or epitope present on the TROP-2 protein.
  • hybridomas expressing anti-TROP-2 monoclonal antibodies can be prepared by immunizing a subject and then isolating hybridomas from the subject’s spleen using routine methods. See, e.g., Milstein et al., (Galfre and Milstein, Methods Enzymol (1981) 73: 3-46). Screening the hybridomas using standard methods will produce monoclonal antibodies of varying specificity (i.e., for different epitopes) and affinity.
  • a selected monoclonal antibody with the desired properties can be used as expressed by the hybridoma, it can be bound to a molecule such as polyethylene glycol (PEG) to alter its properties, or a cDNA encoding it can be isolated, sequenced and manipulated in various ways.
  • Synthetic dendromeric trees can be added to reactive amino acid side chains, e.g., lysine, to enhance the immunogenic properties of TROP-2 protein.
  • CPG-dinucleotide techniques can be used to enhance the immunogenic properties of the TROP-2 protein.
  • the antibody of the present technology is an anti-TROP-2 monoclonal antibody produced by a hybridoma which includes a B cell obtained from a transgenic non-human animal, e.g., a transgenic mouse, having a genome comprising a human heavy chain transgene and a light chain transgene fused to an immortalized cell.
  • Hybridoma techniques include those known in the art and taught in Harlow et al., Antibodies: A Laboratory Manual Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, 349 (1988); Hammerling et al., Monoclonal Antibodies And T-Cell Hybridomas, 563-681 (1981). Other methods for producing hybridomas and monoclonal antibodies are well known to those of skill in the art. [00121] Phage Display Technique. As noted above, the antibodies of the present technology can be produced through the application of recombinant DNA and phage display technology. For example, anti-TROP-2 antibodies, can be prepared using various phage display methods known in -36- 4932-3602-9469.1 Atty. Dkt.
  • phage display methods functional antibody domains are displayed on the surface of a phage particle which carries polynucleotide sequences encoding them.
  • Phages with a desired binding property are selected from a repertoire or combinatorial antibody library (e.g., human or murine) by selecting directly with an antigen, typically an antigen bound or captured to a solid surface or bead.
  • Phages used in these methods are typically filamentous phage including fd and M13 with Fab, Fv or disulfide stabilized Fv antibody domains that are recombinantly fused to either the phage gene III or gene VIII protein.
  • Fab expression libraries See, e.g., Huse, et al., Science 246: 1275-1281, 1989) to allow rapid and effective identification of monoclonal Fab fragments with the desired specificity for a TROP-2 polypeptide, e.g., a polypeptide or derivatives, fragments, analogs or homologs thereof.
  • TROP-2 polypeptide e.g., a polypeptide or derivatives, fragments, analogs or homologs thereof.
  • Other examples of phage display methods that can be used to make the antibodies of the present technology include those disclosed in Huston et al., Proc. Natl. Acad. Sci U.S.A., 85: 5879-5883, 1988; Chaudhary et al., Proc. Natl. Acad.
  • the antibody coding regions from the phage can be isolated and used to generate whole antibodies, including human antibodies, or any other desired antigen binding fragment, and expressed in any desired host including mammalian cells, insect cells, plant cells, yeast, and bacteria.
  • techniques to recombinantly produce Fab, Fab′ and F(ab′) 2 fragments can also be employed using methods known in the art such as those disclosed in WO 92/22324; Mullinax et al., BioTechniques 12: 864-869, 1992; and Sawai et al., AJRI 34: 26-34, 1995; and Better et al., Science 240: 1041- 1043, 1988.
  • hybrid antibodies or hybrid antibody fragments that are cloned into a display vector can be selected against the appropriate antigen in order to identify variants that maintain good binding activity, because the antibody or antibody fragment will be present on the surface of the phage or phagemid particle.
  • a display vector can be selected against the appropriate antigen in order to identify variants that maintain good binding activity, because the antibody or antibody fragment will be present on the surface of the phage or phagemid particle.
  • Barbas III et al. Phage Display, A Laboratory Manual (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 2001).
  • No.115872-3182 could be used for this process, such as cloning the antibody fragment library into a lytic phage vector (modified T7 or Lambda Zap systems) for selection and/or screening.
  • a lytic phage vector modified T7 or Lambda Zap systems
  • the antibodies of the present technology can be produced through the application of recombinant DNA technology.
  • Recombinant polynucleotide constructs encoding an anti-TROP-2 antibody of the present technology typically include an expression control sequence operably-linked to the coding sequences of anti-TROP-2 antibody chains, including naturally-associated or heterologous promoter regions.
  • another aspect of the technology includes vectors containing one or more nucleic acid sequences encoding an anti-TROP-2 antibody of the present technology.
  • the nucleic acid containing all or a portion of the nucleotide sequence encoding the anti-TROP-2 antibody is inserted into an appropriate cloning vector, or an expression vector (i.e., a vector that contains the necessary elements for the transcription and translation of the inserted polypeptide coding sequence) by recombinant DNA techniques well known in the art and as detailed below. Methods for producing diverse populations of vectors have been described by Lerner et al., U.S. Pat.
  • expression vectors useful in recombinant DNA techniques are often in the form of plasmids.
  • plasmid and “vector” can be used interchangeably as the plasmid is the most commonly used form of vector.
  • the present technology is intended to include such other forms of expression vectors that are not technically plasmids, such as viral vectors (e.g., replication defective retroviruses, adenoviruses and adeno-associated viruses), which serve equivalent functions.
  • viral vectors e.g., replication defective retroviruses, adenoviruses and adeno-associated viruses
  • the expression control sequences are eukaryotic promoter systems in vectors capable of transforming or transfecting eukaryotic host cells. Once the vector has been incorporated into the appropriate host, the host is maintained under conditions suitable for high level expression of the nucleotide sequences encoding the anti-TROP-2 antibody, and the collection and purification of the anti-TROP-2 antibody, e.g., cross-reacting anti-TROP-2 antibodies. See generally, U.S.2002/0199213. These expression vectors are typically replicable in the host organisms either as episomes or as an integral part of the host chromosomal DNA.
  • expression vectors contain selection markers, e.g., ampicillin-resistance or hygromycin- resistance, to permit detection of those cells transformed with the desired DNA sequences.
  • Vectors can also encode signal peptide, e.g., pectate lyase, useful to direct the secretion of extracellular antibody fragments. See U.S. Pat. No.5,576,195.
  • the recombinant expression vectors of the present technology comprise a nucleic acid encoding a protein with TROP-2 binding properties in a form suitable for expression of the nucleic acid in a host cell, which means that the recombinant expression vectors include one or more -38- 4932-3602-9469.1 Atty. Dkt.
  • operably-linked is intended to mean that the nucleotide sequence of interest is linked to the regulatory sequence(s) in a manner that allows for expression of the nucleotide sequence (e.g., in an in vitro transcription/translation system or in a host cell when the vector is introduced into the host cell).
  • regulatory sequence is intended to include promoters, enhancers and other expression control elements (e.g., polyadenylation signals).
  • regulatory sequences are described, e.g., in Goeddel, GENE EXPRESSION TECHNOLOGY: METHODS IN ENZYMOLOGY 185, Academic Press, San Diego, Calif. (1990).
  • Regulatory sequences include those that direct constitutive expression of a nucleotide sequence in many types of host cell and those that direct expression of the nucleotide sequence only in certain host cells (e.g., tissue-specific regulatory sequences). It will be appreciated by those skilled in the art that the design of the expression vector can depend on such factors as the choice of the host cell to be transformed, the level of expression of polypeptide desired, etc.
  • Typical regulatory sequences useful as promoters of recombinant polypeptide expression include, e.g., but are not limited to, promoters of 3-phosphoglycerate kinase and other glycolytic enzymes.
  • Inducible yeast promoters include, among others, promoters from alcohol dehydrogenase, isocytochrome C, and enzymes responsible for maltose and galactose utilization.
  • a polynucleotide encoding an anti-TROP-2 antibody of the present technology is operably-linked to an ara B promoter and expressible in a host cell. See U.S. Pat.5,028,530.
  • the expression vectors of the present technology can be introduced into host cells to thereby produce polypeptides or peptides, including fusion polypeptides, encoded by nucleic acids as described herein (e.g., anti-TROP-2 antibody, etc.).
  • Another aspect of the present technology pertains to anti-TROP-2 antibody-expressing host cells, which contain a nucleic acid encoding one or more anti-TROP-2 antibodies.
  • the recombinant expression vectors of the present technology can be designed for expression of an anti- TROP-2 antibody in prokaryotic or eukaryotic cells.
  • an anti-TROP-2 antibody can be expressed in bacterial cells such as Escherichia coli, insect cells (using baculovirus expression vectors), fungal cells, e.g., yeast, yeast cells or mammalian cells. Suitable host cells are discussed further in Goeddel, GENE EXPRESSION TECHNOLOGY: METHODS IN ENZYMOLOGY 185, Academic Press, San Diego, Calif. (1990).
  • the recombinant expression vector can be transcribed and translated in vitro, e.g., using T7 promoter regulatory sequences and T7 polymerase.
  • Fusion vectors add a number of amino acids to a polypeptide encoded therein, usually to the amino terminus of the recombinant polypeptide.
  • Such fusion vectors typically serve three purposes: (i) to increase expression of recombinant polypeptide; (ii) to increase the solubility of the recombinant polypeptide; and (iii) to aid in the purification of the recombinant polypeptide by acting as a ligand in affinity purification.
  • a proteolytic cleavage site is introduced at the junction of the fusion moiety and the recombinant polypeptide to enable separation of the recombinant polypeptide from the fusion moiety subsequent to purification of the fusion polypeptide.
  • enzymes, and their cognate recognition sequences include Factor Xa, thrombin and enterokinase.
  • Typical fusion expression vectors include pGEX (Pharmacia Biotech Inc; Smith and Johnson, 1988.
  • E. coli expression vectors include pTrc (Amrann et al., (1988) Gene 69: 301-315) and pET 11d (Studier et al., GENE EXPRESSION TECHNOLOGY: METHODS IN ENZYMOLOGY 185, Academic Press, San Diego, Calif.
  • the anti-TROP-2 antibody expression vector is a yeast expression vector.
  • yeast expression vectors for expression in yeast Saccharomyces cerevisiae include pYepSec1 (Baldari, et al., 1987.
  • an anti-TROP-2 antibody can be expressed in insect cells using baculovirus expression vectors.
  • a nucleic acid encoding an anti-TROP-2 antibody of the present technology is expressed in mammalian cells using a mammalian expression vector.
  • mammalian expression vectors include, e.g., but are not limited to, pCDM8 (Seed, Nature 329: 840, 1987) and pMT2PC (Kaufman, et al., EMBO J.6: 187-195, 1987).
  • the expression vector's control functions are often provided by viral regulatory elements.
  • promoters are derived from polyoma, adenovirus 2, cytomegalovirus, and simian virus 40.
  • suitable expression systems for both prokaryotic and eukaryotic cells that are useful for expression of the anti-TROP-2 antibody of the present technology, see, e.g., Chapters 16 and 17 of Sambrook, et al., MOLECULAR CLONING: A LABORATORY MANUAL.2nd ed., Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989.
  • the recombinant mammalian expression vector is capable of directing expression of the nucleic acid in a particular cell type (e.g., tissue-specific regulatory elements).
  • tissue-specific regulatory elements are known in the art.
  • suitable tissue-specific promoters include the albumin promoter (liver-specific; Pinkert, et al., Genes Dev.1: 268-277, 1987), lymphoid-specific promoters (Calame and Eaton, Adv.
  • mammary gland-specific promoters e.g., milk whey promoter; U.S. Pat. No. 4,873,316 and European Application Publication No.264,166.
  • Developmentally-regulated promoters are also encompassed, e.g., the murine hox promoters (Kessel and Gruss, Science 249: 374-379, 1990) and the ⁇ -fetoprotein promoter (Campes and Tilghman, Genes Dev.3: 537-546, 1989).
  • Another aspect of the present methods pertains to host cells into which a recombinant expression vector of the present technology has been introduced.
  • a host cell can be any prokaryotic or eukaryotic cell.
  • an anti-TROP-2 antibody can be expressed in bacterial cells such as E. coli, insect cells, yeast or mammalian cells.
  • Expression vectors for these cells can include expression control sequences, such as an origin of replication, a promoter, an enhancer, and necessary processing information sites, such as ribosome binding sites, RNA splice sites, polyadenylation sites, and transcriptional terminator sequences. Queen et al., Immunol. Rev.89: 49, 1986. Illustrative expression control sequences are promoters derived from endogenous genes, cytomegalovirus, SV40, adenovirus, bovine papillomavirus, and the like. Co et al., J Immunol.148: 1149, 1992. Other suitable host cells are known to those skilled in the art.
  • Vector DNA can be introduced into prokaryotic or eukaryotic cells via conventional transformation or transfection techniques.
  • transformation and “transfection” are intended to refer to a variety of art-recognized techniques for introducing foreign nucleic acid (e.g., DNA) into a host cell, including calcium phosphate or calcium chloride co- precipitation, DEAE-dextran-mediated transfection, lipofection, electroporation, biolistics or viral- based transfection.
  • Other methods used to transform mammalian cells include the use of polybrene, protoplast fusion, liposomes, electroporation, and microinjection (See generally, Sambrook et al., Molecular Cloning).
  • Suitable methods for transforming or transfecting host cells can be found in Sambrook, et al. (MOLECULAR CLONING: A LABORATORY MANUAL.2nd ed., Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989), and other laboratory manuals.
  • the vectors containing the DNA segments of interest can be transferred into the host cell by well-known methods, depending on the type of cellular host. [00135]
  • For stable transfection of mammalian cells it is known that, depending upon the expression vector and transfection technique used, only a small fraction of cells may integrate the foreign DNA into their genome.
  • a gene that encodes a selectable marker is generally introduced into the host cells along with the gene of interest.
  • selectable markers include those that confer resistance to drugs, such as G418, hygromycin and methotrexate.
  • Nucleic acid encoding a selectable marker can be introduced into a host cell on the same vector as that encoding the anti-TROP-2 antibody or can be introduced on a separate vector. Cells stably transfected with the introduced nucleic acid can be identified by drug selection (e.g., cells that have incorporated the selectable marker gene will survive, while the other cells die). -42- 4932-3602-9469.1 Atty. Dkt.
  • a host cell that includes an anti-TROP-2 antibody of the present technology can be used to produce (i.e., express) recombinant anti-TROP-2 antibody.
  • the method comprises culturing the host cell (into which a recombinant expression vector encoding the anti-TROP-2 antibody has been introduced) in a suitable medium such that the anti-TROP-2 antibody is produced.
  • the method further comprises the step of isolating the anti-TROP-2 antibody from the medium or the host cell.
  • collections of the anti-TROP-2 antibody e.g., the anti-TROP-2 antibodies or the anti-TROP-2 antibody-related polypeptides are purified from culture media and host cells.
  • the anti-TROP-2 antibody can be purified according to standard procedures of the art, including HPLC purification, column chromatography, gel electrophoresis and the like.
  • the anti-TROP-2 antibody is produced in a host organism by the method of Boss et al., U.S. Pat. No.4,816,397.
  • anti-TROP-2 antibody chains are expressed with signal sequences and are thus released to the culture media.
  • the anti-TROP-2 antibody chains are not naturally secreted by host cells, the anti-TROP-2 antibody chains can be released by treatment with mild detergent.
  • Purification of recombinant polypeptides is well known in the art and includes ammonium sulfate precipitation, affinity chromatography purification technique, column chromatography, ion exchange purification technique, gel electrophoresis and the like (See generally Scopes, Protein Purification (Springer-Verlag, N.Y., 1982).
  • polynucleotides encoding anti-TROP-2 antibodies can be incorporated in transgenes for introduction into the genome of a transgenic animal and subsequent expression in the milk of the transgenic animal. See, e.g., U.S. Pat. Nos. 5,741,957, 5,304,489, and 5,849,992.
  • Suitable transgenes include coding sequences for light and/or heavy chains in operable linkage with a promoter and enhancer from a mammary gland specific gene, such as casein or ⁇ -lactoglobulin.
  • transgenes can be microinjected into fertilized oocytes, or can be incorporated into the genome of embryonic stem cells, and the nuclei of such cells transferred into enucleated oocytes.
  • the anti-TROP-2 antibody of the present technology is a single-chain anti-TROP-2 antibody.
  • techniques can be adapted for the production of single-chain antibodies specific to a TROP-2 protein (See, e.g., U.S. Pat. No.4,946,778). Examples of techniques which can be used to produce single- chain Fvs and antibodies of the present technology include those described in U.S. Pat. Nos.
  • the anti-TROP-2 antibody of the present technology is a chimeric anti-TROP-2 antibody.
  • the anti-TROP-2 -43- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 antibody of the present technology is a humanized anti-TROP-2 antibody.
  • the donor and acceptor antibodies are monoclonal antibodies from different species.
  • the acceptor antibody is a human antibody (to minimize its antigenicity in a human), in which case the resulting CDR-grafted antibody is termed a “humanized” antibody.
  • Recombinant anti-TROP-2 antibodies such as chimeric and humanized monoclonal antibodies, comprising both human and non-human portions, can be made using standard recombinant DNA techniques, and are within the scope of the present technology. For some uses, including in vivo use of the anti-TROP-2 antibody of the present technology in humans as well as use of these agents in in vitro detection assays, it is possible to use chimeric or humanized anti- TROP-2 antibodies.
  • Such chimeric and humanized monoclonal antibodies can be produced by recombinant DNA techniques known in the art. Such useful methods include, e.g., but are not limited to, methods described in International Application No. PCT/US86/02269; U.S. Pat. No. 5,225,539; European Patent No.184187; European Patent No.171496; European Patent No. 173494; PCT International Publication No. WO 86/01533; U.S. Pat. Nos.4,816,567; 5,225,539; European Patent No.125023; Better, et al., 1988. Science 240: 1041-1043; Liu, et al., 1987. Proc. Natl. Acad. Sci.
  • antibodies can be humanized using a variety of techniques including CDR-grafting (EP 0 239400; WO 91/09967; U.S. Pat.
  • a cDNA encoding a murine anti-TROP-2 monoclonal antibody is digested with a restriction enzyme selected specifically to remove the sequence encoding the Fc constant region, and the equivalent portion of a cDNA encoding a human Fc constant region is substituted
  • the present technology provides the construction of humanized anti-TROP-2 antibodies that are unlikely to induce a human anti-mouse antibody (hereinafter referred to as “HAMA”) response, while still having an effective antibody effector function.
  • HAMA human anti-mouse antibody
  • the terms “human” and “humanized”, in relation to antibodies, relate to any antibody which is expected to elicit a therapeutically tolerable weak immunogenic response in a human subject.
  • the present technology provides for a humanized anti-TROP-2 antibodies, heavy and light chain immunoglobulins.
  • CDR Antibodies the anti-TROP-2 antibody of the present technology is an anti-TROP-2 CDR antibody.
  • the donor and acceptor antibodies used to generate the anti-TROP-2 CDR antibody are monoclonal antibodies from different species; typically the acceptor antibody is a human antibody (to minimize its antigenicity in a human), in which case the resulting CDR-grafted antibody is termed a “humanized” antibody.
  • the graft may be of a single CDR (or even a portion of a single CDR) within a single V H or V L of the acceptor antibody, or can be of multiple CDRs (or portions thereof) within one or both of the V H and V L .
  • all three CDRs in all variable domains of the acceptor antibody will be replaced with the corresponding donor CDRs, though one needs to replace only as many as necessary to permit adequate binding of the resulting CDR-grafted antibody to TROP-2 protein.
  • Methods for generating CDR-grafted and humanized antibodies are taught by Queen et al. U.S. Pat. No.5,585,089; U.S. Pat. No.5,693,761; U.S. Pat.
  • DNA sequences encoding the hybrid variable domains described herein can be produced by oligonucleotide synthesis and/or PCR.
  • the nucleic acid encoding CDR regions can also be isolated from the originating species antibodies using suitable restriction enzymes and ligated into the target species framework by ligating with suitable ligation enzymes.
  • the framework regions of the variable chains of the originating species antibody can be changed by site-directed mutagenesis. -45- 4932-3602-9469.1 Atty.
  • hybrids are constructed from choices among multiple candidates corresponding to each framework region, there exist many combinations of sequences which are amenable to construction in accordance with the principles described herein. Accordingly, libraries of hybrids can be assembled having members with different combinations of individual framework regions. Such libraries can be electronic database collections of sequences or physical collections of hybrids. [00145] This process typically does not alter the acceptor antibody’s FRs flanking the grafted CDRs. However, one skilled in the art can sometimes improve antigen binding affinity of the resulting anti-TROP-2 CDR-grafted antibody by replacing certain residues of a given FR to make the FR more similar to the corresponding FR of the donor antibody.
  • Suitable locations of the substitutions include amino acid residues adjacent to the CDR, or which are capable of interacting with a CDR (See, e.g., US 5,585,089, especially columns 12-16). Or one skilled in the art can start with the donor FR and modify it to be more similar to the acceptor FR or a human consensus FR. Techniques for making these modifications are known in the art. Particularly if the resulting FR fits a human consensus FR for that position, or is at least 90% or more identical to such a consensus FR, doing so may not increase the antigenicity of the resulting modified anti-TROP-2 CDR-grafted antibody significantly compared to the same antibody with a fully human FR. [00146] Fc Modifications.
  • the anti-TROP-2 antibodies of the present technology comprise a variant Fc region, wherein said variant Fc region comprises at least one amino acid modification relative to a wild-type Fc region (or the parental Fc region), such that said molecule has an altered affinity for an Fc receptor (e.g., an Fc ⁇ R), provided that said variant Fc region does not have a substitution at positions that make a direct contact with Fc receptor based on crystallographic and structural analysis of Fc-Fc receptor interactions such as those disclosed by Sondermann et al., Nature, 406:267-273 (2000).
  • an Fc receptor e.g., an Fc ⁇ R
  • an anti-TROP-2 antibody of the present technology has an altered affinity for activating and/or inhibitory receptors, having a variant Fc region with one or more amino acid modifications, wherein said one or more amino acid modification is a N297 substitution with alanine, or a K322 substitution with alanine.
  • anti-TROP-2 antibodies of the present technology have an Fc region with variant glycosylation as compared to a parent Fc region.
  • variant glycosylation includes the absence of fucose; in some embodiments, variant glycosylation results from expression in GnT1-deficient CHO cells. -46- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 [00149]
  • the antibodies of the present technology may have a modified glycosylation site relative to an appropriate reference antibody that binds to an antigen of interest (e.g., TROP-2), without altering the functionality of the antibody, e.g., binding activity to the antigen.
  • an antigen of interest e.g., TROP-2
  • glycosylation sites include any specific amino acid sequence in an antibody to which an oligosaccharide (i.e., carbohydrates containing two or more simple sugars linked together) will specifically and covalently attach.
  • Oligosaccharide side chains are typically linked to the backbone of an antibody via either N-or O-linkages.
  • N-linked glycosylation refers to the attachment of an oligosaccharide moiety to the side chain of an asparagine residue.
  • O-linked glycosylation refers to the attachment of an oligosaccharide moiety to a hydroxyamino acid, e.g., serine, threonine.
  • an Fc- glycoform (hTROP-2-IgGln) that lacks certain oligosaccharides including fucose and terminal N- acetylglucosamine may be produced in special CHO cells and exhibit enhanced ADCC effector function.
  • the carbohydrate content of an immunoglobulin-related composition disclosed herein is modified by adding or deleting a glycosylation site. Methods for modifying the carbohydrate content of antibodies are well known in the art and are included within the present technology, see, e.g., U.S. Patent No.6,218,149; EP 0359096B1; U.S. Patent Publication No.
  • the carbohydrate content of an antibody is modified by deleting one or more endogenous carbohydrate moieties of the antibody.
  • the present technology includes deleting the glycosylation site of the Fc region of an antibody, by modifying position 297 from asparagine to alanine.
  • Engineered glycoforms may be useful for a variety of purposes, including but not limited to enhancing or reducing effector function.
  • Engineered glycoforms may be generated by any method known to one skilled in the art, for example by using engineered or variant expression strains, by co-expression with one or more enzymes, for example N-acetylglucosaminyltransferase III (GnTIII), by expressing a molecule comprising an Fc region in various organisms or cell lines from various organisms, or by modifying carbohydrate(s) after the molecule comprising Fc region has been expressed.
  • Methods for generating engineered glycoforms are known in the art, and include but are not limited to those described in Umana et al., 1999, Nat.
  • the anti-TROP-2 antibody of the present technology is a fusion protein.
  • the anti-TROP-2 antibodies of the present technology when fused to a second protein, can be used as an antigenic tag.
  • domains that can be fused to polypeptides include not only heterologous signal sequences, but also other heterologous functional regions.
  • the fusion does not necessarily need to be direct, but can occur through linker sequences.
  • fusion proteins of the present technology can also be engineered to improve characteristics of the anti-TROP-2 antibodies. For instance, a region of additional amino acids, particularly charged amino acids, can be added to the N-terminus of the anti-TROP-2 antibody to improve stability and persistence during purification from the host cell or subsequent handling and storage.
  • peptide moieties can be added to an anti-TROP-2 antibody to facilitate purification. Such regions can be removed prior to final preparation of the anti-TROP-2 antibody.
  • the addition of peptide moieties to facilitate handling of polypeptides are familiar and routine techniques in the art.
  • the anti-TROP-2 antibody of the present technology can be fused to marker sequences, such as a peptide which facilitates purification of the fused polypeptide.
  • the marker amino acid sequence is a hexa-histidine peptide, such as the tag provided in a pQE vector (QIAGEN, Inc., Chatsworth, Calif), among others, many of which are commercially available. As described in Gentz et al., Proc. Natl.
  • hexa-histidine provides for convenient purification of the fusion protein.
  • Another peptide tag useful for purification, the “HA” tag corresponds to an epitope derived from the influenza hemagglutinin protein. Wilson et al., Cell 37: 767, 1984.
  • any of these above fusion proteins can be engineered using the polynucleotides or the polypeptides of the present technology.
  • the fusion proteins described herein show an increased half-life in vivo.
  • Fusion proteins having disulfide-linked dimeric structures can be more efficient in binding and neutralizing other molecules compared to the monomeric secreted protein or protein fragment alone.
  • EP-A-O 464533 (Canadian counterpart 2045869) discloses fusion proteins comprising various portions of constant region of immunoglobulin molecules together with another human protein or a fragment thereof. In many cases, the Fc part in a fusion protein is beneficial in -48- 4932-3602-9469.1 Atty. Dkt.
  • No.115872-3182 therapy and diagnosis can result in, e.g., improved pharmacokinetic properties. See EP-A 0232262.
  • deleting or modifying the Fc part after the fusion protein has been expressed, detected, and purified may be desired.
  • the Fc portion can hinder therapy and diagnosis if the fusion protein is used as an antigen for immunizations.
  • human proteins, such as hTROP-2 have been fused with Fc portions for the purpose of high- throughput screening assays to identify antagonists of hTROP-2. Bennett et al., J. Molecular Recognition 8: 52-58, 1995; Johanson et al., J. Biol.
  • the anti-TROP-2 antibody of the present technology is coupled with a label moiety, i.e., detectable group.
  • the particular label or detectable group conjugated to the anti-TROP-2 antibody is not a critical aspect of the technology, so long as it does not significantly interfere with the specific binding of the anti-TROP-2 antibody of the present technology to the TROP-2 protein.
  • the detectable group can be any material having a detectable physical or chemical property. Such detectable labels have been well-developed in the field of immunoassays and imaging. In general, almost any label useful in such methods can be applied to the present technology.
  • a label is any composition detectable by spectroscopic, photochemical, biochemical, immunochemical, electrical, optical or chemical means.
  • Labels useful in the practice of the present technology include magnetic beads (e.g., DynabeadsTM), fluorescent dyes (e.g., fluorescein isothiocyanate, Texas red, rhodamine, and the like), radiolabels (e.g., 3 H, 14 C, 35 S, 125 I, 121 I, 131 I, 112 In, 99 mTc), other imaging agents such as microbubbles (for ultrasound imaging), 18 F, 11 C, 15 O, 89 Zr (for Positron emission tomography), 99m TC, 111 In (for Single photon emission tomography), enzymes (e.g., horse radish peroxidase, alkaline phosphatase and others commonly used in an ELISA), and calorimetric labels such as colloidal gold or colored glass or plastic (e.g., polystyrene, poly
  • Non-radioactive labels are often attached by indirect means.
  • a ligand molecule e.g., biotin
  • the ligand then binds to an anti-ligand (e.g., streptavidin) molecule which is either inherently detectable or covalently bound to a signal system, such as a detectable enzyme, a fluorescent compound, or a chemiluminescent compound.
  • an anti-ligand e.g., streptavidin
  • a number of ligands and anti-ligands can be used. Where a ligand has a natural anti-ligand, e.g., -49- 4932-3602-9469.1 Atty. Dkt.
  • biotin, thyroxine, and cortisol it can be used in conjunction with the labeled, naturally-occurring anti-ligands.
  • any haptenic or antigenic compound can be used in combination with an antibody, e.g., an anti-TROP-2 antibody.
  • the molecules can also be conjugated directly to signal generating compounds, e.g., by conjugation with an enzyme or fluorophore. Enzymes of interest as labels will primarily be hydrolases, particularly phosphatases, esterases and glycosidases, or oxidoreductases, particularly peroxidases.
  • Fluorescent compounds useful as labeling moieties include, but are not limited to, e.g., fluorescein and its derivatives, rhodamine and its derivatives, dansyl, umbelliferone, and the like.
  • Chemiluminescent compounds useful as labeling moieties include, but are not limited to, e.g., luciferin, and 2,3-dihydrophthalazinediones, e.g., luminol.
  • Means of detecting labels are well known to those of skill in the art.
  • means for detection include a scintillation counter or photographic film as in autoradiography.
  • the label is a fluorescent label
  • it can be detected by exciting the fluorochrome with the appropriate wavelength of light and detecting the resulting fluorescence.
  • the fluorescence can be detected visually, by means of photographic film, by the use of electronic detectors such as charge coupled devices (CCDs) or photomultipliers and the like.
  • enzymatic labels can be detected by providing the appropriate substrates for the enzyme and detecting the resulting reaction product.
  • simple colorimetric labels can be detected simply by observing the color associated with the label.
  • conjugated gold often appears pink, while various conjugated beads appear the color of the bead.
  • Some assay formats do not require the use of labeled components.
  • agglutination assays can be used to detect the presence of the target antibodies, e.g., the anti-TROP- 2 antibodies.
  • antigen-coated particles are agglutinated by samples comprising the target antibodies.
  • none of the components need be labeled and the presence of the target antibody is detected by simple visual inspection.
  • the immunoglobulin-related compositions (e.g., antibodies or antigen binding fragments thereof) of the present technology are useful for treating diffuse pleural mesothelioma (DPM) in a subject in need thereof comprising administering to the subject an effective amount of an antibody-drug conjugate comprising an anti-TROP-2 antibody or antigen binding fragment thereof.
  • the subject is -50- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 diagnosed with or is suffering from recurrent DPM or metastatic DPM.
  • the subject has received a prior anti-cancer therapy.
  • the present disclosure provides a method for treating diffuse pleural mesothelioma (DPM) in a subject in need thereof comprising administering to the subject an effective amount of an antibody-drug conjugate comprising an anti-TROP-2 antibody or antigen binding fragment thereof and an effective amount of an AKT inhibitor.
  • the present disclosure provides a method for re-sensitizing DPM tumors to anti-TROP-2 antibody-drug conjugate therapy comprising administering to the subject an effective amount of an antibody-drug conjugate comprising an anti-TROP-2 antibody or antigen binding fragment thereof and an effective amount of an AKT inhibitor.
  • AKT inhibitors include, but are not limited to, samotolisib, oridonin, capivasertib, ipatasertib, miransertib, afuresertib, uprosertib, BAY1125976, MK-2206, TAS-117, GSK690693, triciribine, and perifosine.
  • the antibody-drug conjugate and the AKT inhibitor are administered separately, simultaneously, or sequentially.
  • the antibody-drug conjugate comprises one or more of alkylating agents, topoisomerase inhibitors, platinum agents, taxanes, vinca agents, anti-estrogen drugs, aromatase inhibitors, ovarian suppression agents, VEGF/VEGFR inhibitors, EGF/EGFR inhibitors, PARP inhibitors, cytostatic alkaloids, cytotoxic antibiotics, antimetabolites, endocrine/hormonal agents, and bisphosphonate therapy agents.
  • the antibody-drug conjugate comprises a cytotoxic drug selected from the group consisting of an anthracycline, a camptothecin, a tubulin inhibitor, a maytansinoid, a calicheamycin, an auristatin, a nitrogen mustard, an ethylenimine derivative, an alkyl sulfonate, a nitrosourea, a triazene, a folic acid analog, a taxane, a COX-2 inhibitor, a pyrimidine analog, a purine analog, an antibiotic, an enzyme inhibitor, an epipodophyllotoxin, a platinum coordination complex, a vinca alkaloid, a substituted urea, a methyl hydrazine derivative, an adrenocortical suppressant, a hormone antagonist, an antimetabolite, an alkylating agent, an antimitotic, an anti-angiogenic agent, a tyrosine kinase inhibitor,
  • the antibody-drug conjugate comprises cyclophosphamide, fluorouracil (or 5-fluorouracil or 5-FU), methotrexate, edatrexate (10- -51- 4932-3602-9469.1 Atty. Dkt.
  • the anti-TROP-2 antibody or antigen binding fragment comprises the variable heavy domain and variable light domain of sacituzumab, or datopotamab.
  • the antibody-drug conjugate comprises sacituzumab govitecan.
  • the compositions of the present technology may optionally be administered as a single bolus to a subject in need thereof.
  • the dosing regimen may comprise multiple administrations performed at various times after the appearance of tumors. -52- 4932-3602-9469.1 Atty. Dkt.
  • Administration can be carried out by any suitable route, including orally, intranasally, parenterally (intravenously, intramuscularly, intraperitoneally, or subcutaneously), rectally, intracranially, intratumorally, intrathecally, or topically. Administration includes self-administration and the administration by another. It is also to be appreciated that the various modes of treatment of medical conditions as described are intended to mean “substantial”, which includes total but also less than total treatment, and wherein some biologically or medically relevant result is achieved.
  • the methods of the present technology further comprise sequentially, simultaneously or separately administering to the patient an effective amount of a corticosteroid, or an antihistamine.
  • the antibodies of the present technology comprise pharmaceutical formulations which may be administered to subjects in need thereof in one or more doses. Dosage regimens can be adjusted to provide the desired response (e.g., a therapeutic response).
  • an effective amount of the antibody compositions of the present technology sufficient for achieving a therapeutic effect, range from about 0.000001 mg per kilogram body weight per day to about 10,000 mg per kilogram body weight per day.
  • the dosage ranges are from about 0.0001 mg per kilogram body weight per day to about 100 mg per kilogram body weight per day.
  • the dosage ranges from about 0.0001 to 100 mg/kg, and more usually 0.01 to 5 mg/kg every week, every two weeks or every three weeks, of the subject body weight.
  • dosages can be 1 mg/kg body weight or 10 mg/kg body weight every week, every two weeks or every three weeks or within the range of 1-10 mg/kg every week, every two weeks or every three weeks.
  • a single dosage of antibody ranges from 0.1-10,000 micrograms per kg body weight.
  • antibody concentrations in a carrier range from 0.2 to 2000 micrograms per delivered milliliter.
  • An exemplary treatment regime entails administration once per every two weeks or once a month or once every 3 to 6 months.
  • Anti-TROP-2 antibodies may be administered on multiple occasions. Intervals between single dosages can be hourly, daily, weekly, monthly or yearly. Intervals can also be irregular as indicated by measuring blood levels of the antibody in the subject.
  • dosage is adjusted to achieve a serum antibody concentration in the subject of from about 75 ⁇ g/mL to about 125 ⁇ g/mL, 100 ⁇ g/mL to about 150 ⁇ g/mL, from about 125 ⁇ g/mL to about 175 ⁇ g/mL, or from about 150 ⁇ g/mL to about 200 ⁇ g/mL.
  • anti-TROP-2 antibodies can be administered as a sustained release formulation, in which case less frequent administration is required. Dosage and frequency vary depending on the half-life of the antibody in the subject. The dosage and frequency of administration can vary depending on whether the treatment is prophylactic or therapeutic. In prophylactic applications, a relatively low dosage is administered at relatively infrequent intervals over a long period of time. In therapeutic applications, a relatively high dosage at relatively short intervals is sometimes required until progression of the disease is reduced or terminated, or until the subject shows partial or complete amelioration of symptoms of disease. Thereafter, the patient can be administered a prophylactic regime. [00173] Toxicity.
  • an effective amount (e.g., dose) of an anti-TROP-2 antibody described herein will provide therapeutic benefit without causing substantial toxicity to the subject.
  • Toxicity of the anti-TROP-2 antibody described herein can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., by determining the LD50 (the dose lethal to 50% of the population) or the LD100 (the dose lethal to 100% of the population). The dose ratio between toxic and therapeutic effect is the therapeutic index. The data obtained from these cell culture assays and animal studies can be used in formulating a dosage range that is not toxic for use in human.
  • the dosage of the anti-TROP-2 antibody described herein lies within a range of circulating concentrations that include the effective dose with little or no toxicity.
  • the dosage can vary within this range depending upon the dosage form employed and the route of administration utilized. The exact formulation, route of administration and dosage can be chosen by the individual physician in view of the subject’s condition. See, e.g., Fingl et al., In: The Pharmacological Basis of Therapeutics, Ch.1 (1975).
  • Formulations of Pharmaceutical Compositions According to the methods of the present technology, the anti-TROP-2 antibody can be incorporated into pharmaceutical compositions suitable for administration.
  • the pharmaceutical compositions generally comprise recombinant or substantially purified antibody and a pharmaceutically- acceptable carrier in a form suitable for administration to a subject. Pharmaceutically- acceptable carriers are determined in part by the particular composition being administered, as well as by the particular method used to administer the composition.
  • compositions for administering the antibody compositions See, e.g., Remington’s Pharmaceutical Sciences, Mack Publishing Co., Easton, PA 18 th ed., 1990).
  • the pharmaceutical compositions are generally -54- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 formulated as sterile, substantially isotonic and in full compliance with all Good Manufacturing Practice (GMP) regulations of the U.S. Food and Drug Administration.
  • GMP Good Manufacturing Practice
  • compositions, carriers, diluents and reagents are used interchangeably and represent that the materials are capable of administration to or upon a subject without the production of undesirable physiological effects to a degree that would prohibit administration of the composition.
  • pharmaceutically- acceptable excipient means an excipient that is useful in preparing a pharmaceutical composition that is generally safe, non-toxic, and desirable, and includes excipients that are acceptable for veterinary use as well as for human pharmaceutical use. Such excipients can be solid, liquid, semisolid, or, in the case of an aerosol composition, gaseous.
  • “Pharmaceutically-acceptable salts and esters” means salts and esters that are pharmaceutically-acceptable and have the desired pharmacological properties. Such salts include salts that can be formed where acidic protons present in the composition are capable of reacting with inorganic or organic bases. Suitable inorganic salts include those formed with the alkali metals, e.g., sodium and potassium, magnesium, calcium, and aluminum. Suitable organic salts include those formed with organic bases such as the amine bases, e.g., ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, and the like.
  • Such salts also include acid addition salts formed with inorganic acids (e.g., hydrochloric and hydrobromic acids) and organic acids (e.g., acetic acid, citric acid, maleic acid, and the alkane- and arene-sulfonic acids such as methanesulfonic acid and benzenesulfonic acid).
  • Pharmaceutically-acceptable esters include esters formed from carboxy, sulfonyloxy, and phosphonoxy groups present in the anti-TROP-2 antibody, e.g., C1-6 alkyl esters.
  • a pharmaceutically-acceptable salt or ester can be a mono-acid-mono-salt or ester or a di-salt or ester; and similarly where there are more than two acidic groups present, some or all of such groups can be salified or esterified.
  • An anti-TROP-2 antibody named in this technology can be present in unsalified or unesterified form, or in salified and/or esterified form, and the naming of such anti- TROP-2 antibody is intended to include both the original (unsalified and unesterified) compound and its pharmaceutically-acceptable salts and esters.
  • certain embodiments of the present technology can be present in more than one stereoisomeric form, and the naming of such anti-TROP-2 antibody is intended to include all single stereoisomers and all mixtures (whether racemic or otherwise) of such stereoisomers.
  • Examples of such carriers or diluents include, but are not limited to, water, saline, Ringer's solutions, dextrose solution, and 5% human serum albumin.
  • a pharmaceutical composition of the present technology is formulated to be compatible with its intended route of administration.
  • the anti-TROP-2 antibody compositions of the present technology can be administered by parenteral, topical, intravenous, oral, subcutaneous, intraarterial, intradermal, transdermal, rectal, intracranial, intrathecal, intraperitoneal, intranasal; or intramuscular routes, or as inhalants.
  • Solutions or suspensions used for parenteral, intradermal, or subcutaneous application can include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial compounds such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating compounds such as ethylenediaminetetraacetic acid (EDTA); buffers such as acetates, citrates or phosphates, and compounds for the adjustment of tonicity such as sodium chloride or dextrose.
  • a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents
  • antibacterial compounds such as benzyl alcohol or methyl parabens
  • antioxidants such as ascorbic acid or sodium bisulfite
  • chelating compounds
  • compositions suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion.
  • suitable carriers include physiological saline, bacteriostatic water, Cremophor EL TM (BASF, Parsippany, N.J.) or phosphate buffered saline (PBS).
  • the carrier can be a solvent or dispersion medium containing, e.g., water, ethanol, polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof.
  • the proper fluidity can be maintained, e.g., by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants.
  • a coating such as lecithin
  • surfactants e.g., sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium bicarbonate, sodium sorbitol, sodium chloride in the composition.
  • Prolonged absorption of the injectable compositions can be brought about by including in the composition a compound which delays absorption, e.g., aluminum monostearate and gelatin.
  • Sterile injectable solutions can be prepared by incorporating an anti-TROP-2 antibody of the present technology in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by filtered sterilization.
  • dispersions are prepared by incorporating the anti-TROP-2 antibody into a sterile vehicle that contains a basic dispersion medium and the required other ingredients from those enumerated above.
  • Oral compositions can also be prepared using a fluid carrier for use as a mouthwash, wherein the compound in the fluid carrier is applied orally and swished and expectorated or swallowed.
  • Pharmaceutically compatible binding compounds, and/or adjuvant materials can be included as part of the composition.
  • the tablets, pills, capsules, troches and the like can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating compound such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium -57- 4932-3602-9469.1 Atty. Dkt.
  • the anti-TROP-2 antibody is delivered in the form of an aerosol spray from pressured container or dispenser which contains a suitable propellant, e.g., a gas such as carbon dioxide, or a nebulizer.
  • a suitable propellant e.g., a gas such as carbon dioxide, or a nebulizer.
  • Systemic administration can also be by transmucosal or transdermal means.
  • penetrants appropriate to the barrier to be permeated are used in the formulation.
  • Such penetrants are generally known in the art, and include, e.g., for transmucosal administration, detergents, bile salts, and fusidic acid derivatives. Transmucosal administration can be accomplished through the use of nasal sprays or suppositories.
  • the anti-TROP-2 antibody is formulated into ointments, salves, gels, or creams as generally known in the art.
  • the anti-TROP-2 antibody can also be prepared as pharmaceutical compositions in the form of suppositories (e.g., with conventional suppository bases such as cocoa butter and other glycerides) or retention enemas for rectal delivery.
  • the anti-TROP-2 antibody is prepared with carriers that will protect the anti-TROP-2 antibody against rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems.
  • a controlled release formulation including implants and microencapsulated delivery systems.
  • Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Methods for preparation of such formulations will be apparent to those skilled in the art.
  • the materials can also be obtained commercially from Alza Corporation and Nova Pharmaceuticals, Inc.
  • Liposomal suspensions (including liposomes targeted to infected cells with monoclonal antibodies to viral antigens) can also be used as pharmaceutically- acceptable carriers.
  • kits for the treatment of DPM comprising at least one immunoglobulin-related composition of the present technology (e.g., any antibody or antigen binding fragment described herein), or a functional variant (e.g., substitutional variant) thereof.
  • the above described components of the kits of the present technology are packed in suitable containers and labeled for treatment of DPM.
  • No.115872-3182 mentioned components may be stored in unit or multi-dose containers, for example, sealed ampoules, vials, bottles, syringes, and test tubes, as an aqueous, preferably sterile, solution or as a lyophilized, preferably sterile, formulation for reconstitution.
  • the kit may further comprise a second container which holds a diluent suitable for diluting the pharmaceutical composition towards a higher volume. Suitable diluents include, but are not limited to, the pharmaceutically acceptable excipient of the pharmaceutical composition and a saline solution.
  • the kit may comprise instructions for diluting the pharmaceutical composition and/or instructions for administering the pharmaceutical composition, whether diluted or not.
  • the containers may be formed from a variety of materials such as glass or plastic and may have a sterile access port (for example, the container may be an intravenous solution bag or a vial having a stopper which may be pierced by a hypodermic injection needle).
  • the kit may further comprise more containers comprising a pharmaceutically acceptable buffer, such as phosphate-buffered saline, Ringer's solution and dextrose solution. It may further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles, syringes, culture medium for one or more of the suitable hosts.
  • kits may optionally include instructions customarily included in commercial packages of therapeutic or diagnostic products, that contain information about, for example, the indications, usage, dosage, manufacture, administration, contraindications and/or warnings concerning the use of such therapeutic or diagnostic products.
  • the kit can also comprise, e.g., a buffering agent, a preservative or a protein- stabilizing agent.
  • Each component of the kit can be enclosed within an individual container and all of the various containers can be within a single package, along with instructions for using the kit.
  • the kits of the present technology may contain a written product on or in the kit container. The written product describes how to use the reagents contained in the kit, e.g., for treatment of DPM in a subject in need thereof.
  • TROP-2 expression was analyzed in independent DPM clinical cohorts at the mRNA and protein levels and correlated to different clinical features including patient survival. Isogenic cell lines were generated to perform a functional study of the effect of the modulation of TROP-2 expression on tumorigenic characteristics in vitro and in vivo. An array of patient-derived xenograft models were treated with a TROP-2-targeting antibody-drug conjugate in vivo to assess its efficacy at controlling tumor growth, and compare it to that of standard of care cytotoxin therapy. Endpoints or each treatment group were defined as the time when group average size would reach 1000mm 3 of volume. Where specified, tumors were collected, and molecular analyses were performed on then to identify and validate mechanisms of resistance to treatment.
  • Plasmid vectors, virus production and transductions Lentiviral particles were produced and used to infect isogenic cell lines of interest, as described previously (50).
  • Lentiviral vectors used include Cas9 (#125592, Addgene, RRID:Addgene_125592), LV04 vectors expressing sgRNAs for CDC7 (#HSPD0000024539, Sigma) or the respective control vector expressing a safe targeting sgRNA BFP (#HSCONTROL_AAVS1 on LV04, Sigma), or the Lv105 vector overexpressing TACSTD2 (#EX- G0457-Lv105-B Genecopoeia). No clone isolation/expansion was doing after transduction with either Cas9, -60- 4932-3602-9469.1 Atty. Dkt.
  • Lentiviral particles were produced and used to infect isogenic cell lines of interest, as described previously (57), through concurrent transfection of HEK293T cells (ATCC, # CRL-1573, RRID:CVCL_0045) with a 3:2:1 ratio of lentiviral plasmid:psPAX2:pMD2.G with JetPrime transfection reagent (Polyplus, # 114-15) at a 2:1 JetPrime:DNA ratio. Medium was changed 24 h after transfection and viral supernatants were collected 72 h after transfection.
  • Viral supernatants were syringe-filtered with a 0.45- ⁇ M PVDF filter (Millipore, # SLHVM33RS) and concentrated approximately 20-fold with Lenti-X Concentrator (Takara Bio, # 631232) according to the manufacturer’s protocol.
  • lentiviral particles made out of a lentiviral plasmid designed to constitutively express Cas9 (#125592, Addgene, RRID:Addgene_125592) as described in (58), and selected with blasticidin 2.5 ⁇ g/mL.
  • Cells were similarly spin-transduced as described in (1) with lentiviral particles made out of lentiviral LV04 vectors expressing sgRNAs for CDC7 (#HSPD0000024539, Sigma) or the respective control vector expressing a safe targeting sgRNA BFP (#HSCONTROL_AAVS1 on LV04, Sigma), or the Lv105 vector overexpressing TACSTD2 (#EX-G0457-Lv105-B Genecopoeia).
  • Synergy assays Cells were seeded in 96-well plates (1000 cells/well) and treated with an interval of concentrations of SG and samotolisib (Selleck Chemicals, #S8322) for 5 days.
  • Protein extraction and western blot were performed as previously described in (59) from frozen cell pellets or flash-frozen tumor samples using RIPA lysis buffer with 1 ⁇ HALT protease inhibitor cocktail (Thermo, # 78446). Cell pellets were resuspended in five volumes of cold lysis buffer and incubated on ice for 30 min. Lysates were clarified by centrifugation at 20,000g for 10 min at 4 °C. Antibodies for TROP-2 (Cell Signaling -61- 4932-3602-9469.1 Atty. Dkt.
  • IHC was performed on FFPE tissue from tumor samples or PDX slides or tissue microarrays derived from patients with DPM, using a TROP-2 antibody (#NBP2-49166, Novus Biologicals).
  • TROP-2 antibody #NBP2-49166, Novus Biologicals
  • slides were deparaffinized and steamed for 45 min in Target Retrieval Solution (Dako).
  • Immunocomplexes were detected using PV Poly-HRP anti-mouse IgG (Leica Microsystems, #PV6114) followed by a TSA biotin amplification step (Perkin Elmer) with DAB as the chromogen.
  • Tissue sections were counterstained with hematoxylin, and slides were digitized on a Ventana DP 200 Slide Scanner (Roche). Expression was scored in a blinded manner by pathologists, whereby the optical density level (“0” for no brown color, “1” for faint and fine brown chromogen deposition, “2” for intermediate chromogen deposition, and “3” for prominent chromogen deposition) was multiplied by the percentage of cells at each staining level, resulting in a total H-score range of 0–300.
  • mice were kept in specific pathogen-free animal facilities at Memorial Sloan Kettering Cancer Center (MSK), and procedures were performed in accord with the guidelines of MSK Institutional Animal Care and Use Committee under an approved protocol.
  • MSK Memorial Sloan Kettering Cancer Center
  • Six to ten female 6-week-old NOD.Cg- Prkdc ⁇ scid> Il2rg ⁇ tm1Wjl>/SzJ (NSG) mice were subcutaneously engrafted per treatment arm and until tumors reached 100-150 mm 3 . At that point, mice were randomized into groups and treated with either vehicle, SG (0.5 mg/mouse i.v. BIW), gemcitabine (40mg/kg -62- 4932-3602-9469.1 Atty. Dkt.
  • SG 0.5 mg/mouse i.v. BIW
  • gemcitabine 40mg/kg -62- 4932-3602-9469.1 Atty. Dkt.
  • mice No.115872-3182 i.p. QW (53-55) or irinotecan (50 mg/kg i.p QW), or with the combination of SG (0.5 mg/mouse i.v. BIW) and samotolosib (5 mg/kg p.o. QDx5).
  • Mice weights and tumor volumes were measured twice a week and mice were sacrificed when tumors reached ⁇ 1000 mm 3 . The number of mice per treatment arm were selected according to previous experience with the models and response to treatments. Blinding was not performed. All animal experiments were approved by the MSK Institutional Animal Care and Use Committee (#13-07-007).
  • RNA seq analyses For PDX samples the FASTQ files were first mapped to a hybrid genome that consists of both human and mouse sequences into one index. The reads were mapped and then any read that mapped to the mouse genome was culled. The remaining reads were converted back to a FASTQ file and mapped to the target genome using the rnaStar aligner that maps reads genomically and resolves reads across splice junctions (61).
  • the 2-pass mapping method was used in which the reads are mapped twice (62).
  • the first mapping pass used a list of known annotated junctions from Ensemble. Novel junctions found in the first pass were then added to the known junctions and a second mapping pass was done (on the second pass the RemoveNoncanoncial flag was used).
  • the output SAM files were post-processed using the PICARD tools to: add read groups, AddOrReplaceReadGroups which in addition sorted the file and converted it to the compressed BAM format.
  • the expression count matrix was then computed from the mapped reads using HTSeq (www-huber.embl.de/users/anders/HTSeq, RRID:SCR_005514) and one of several possible gene model databases.
  • transcript counts were quantified using Salmon v1.1.0 (RRID:SCR_016863)(66) from RNA-seq reads. Fastq raw reads were mapped to 25 mer indexed hg38 genome by Salmon with default settings. Transcripts were converted to genes based on Ensembl 92 (RRID:SCR_002344)(67) and normalized by size factor at gene level. The subsequent differential gene expression analyses were processed on Salmon output files using Sleuth v0.30.0 (RRID:SCR_016883) in gene mode (68). Differentially expressed genes were identified using the Wald test.
  • GSEA Gene set enrichment analysis
  • MSK_Lx13 tumor bearing mice -65- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 were injected with 20 ⁇ g [ 89 Zr]Zr-DFO-SG and imaged on a Siemens Inveon PET/CT system at 24-, 48-, and 72-hours post injection, while mice bearing MSK_Lx606 tumors were imaged at 24-, 48-, and 120-hours post injection and images calibrated to the percent injected dose per gram.
  • mice Post imaging, mice were euthanized, and the following organs harvested for gamma counting and weighing as an orthogonal method to imaging blood, heart, lung, liver, spleen, pancreas, small intestine, large intestine, kidneys, muscle, bone, and tumor.
  • Lx606 mouse model a blocking study was performed with 500 ⁇ g IgG 1 pre mouse alongside the 20 ⁇ g [ 89 Zr]Zr-DFO-SG.
  • Statistical analyses Comparisons between two groups were performed using paired or unpaired two-tailed Student’s t test, as described in the figure legends.
  • RNA extraction Frozen tissues or cell pellets were weighed and homogenized in RLT and nucleic acids were extracted using the AllPrep DNA/RNA Mini Kit (QIAGEN, #80204) according to the manufacturer’s instructions.
  • TPM and STAR-Counts TCGA expression matrices (TPM and STAR-Counts) and corresponding meta data for the “TCGA-MESO” project were downloaded using the “TCGAbiolinks” R package v2.30.4 (The Cancer Genome Atlas (RRID:SCR_003193))(70-72). Samples were divided into thirds based on expression of TACSTD2 (TPM quantification). Differential gene expression analysis (DEG) was performed between the highest and lowest groups using the DESeq2 R package (RRID:SCR_015687) with the STAR-Counts expression data.
  • DEG Differential gene expression analysis
  • Genes were ranked according to a composite score (-log10(pvalue)*sign(log2FoldChange)) and gene set enrichment analysis was performed with the ⁇ clusterProfiler ⁇ R package using gene sets from MSigDB (RRID:SCR_016863), Kegg (https://www.genome.jp/kegg/, SCR_012773) and Gene Ontology (GO) (https://geneontology.org/, RRID:SCR_002811). Pathways were filtered for p ⁇ 0.05 and manually curated based on pathway among all gene sets. Pathways -66- 4932-3602-9469.1 Atty. Dkt.
  • TROP-2 expression in DPM clinical samples were performed similarly to the TCGA cohort.
  • IHC immunohistochemistry
  • TMA patient-derived DPM tissue microarray
  • 20 benign pleural specimens submitted for TROP-2 IHC evaluation exhibited no detectable TROP-2 expression, suggesting that TROP-2 expression appears limited to tumor tissue in this setting (p ⁇ 0.0001; FIG.1A).
  • TROP-2 expression was associated with poor prognosis in DPM.
  • TROP-2 activates oncogenic and pro-metastatic signaling in DPM
  • TCGA Cancer Genome Atlas
  • MESOMICS MESOMICS
  • Differential gene expression (DGE) compared tumors with high (highest tertile) versus low (lowest tertile) TACSTD2 mRNA expression, followed by pathway enrichment analyses, to nominate pathways potentially dysregulated by TROP-2 expression (FIGs.2A-2B).
  • TROP-2 high- -67- 4932-3602-9469.1 Atty. Dkt.
  • SG efficacy was tested in MSK_Lx707 (treatment naive, low TROP-2 expression), MSK_Lx13 (pre-treated, low expression), MSK_Lx307 (treatment naive, intermediate expression) and MSK_Lx606 (pre-treated, high TROP-2 expression) (FIG.4A) (15).
  • SG showed efficacy superior to gemcitabine in all models, regardless of TROP-2 expression or pre-treatment status (FIG.4B), with treatment/control (T/C) values of 14% vs.72% (MSK_Lx707), 32% vs.45% (MSK_Lx307) and 10% vs.
  • SG The efficacy of SG was also compared with that of irinotecan (the active metabolite of the SG payload, SN38) in the two PDX models derived from pre- treated DPM tumors (FIG.4B). Notably, irinotecan showed minimal efficacy, suggesting that ADC-driven internalization may be important for response. SG appeared to be well tolerated in all PDX models, including no significant decrease in body weight relative to the controls (FIG.8A). These results suggest that SG might be an attractive therapeutic approach for patients with recalcitrant DPM tumors.
  • mice bearing the SG- sensitive model MSK_Lx606 were treated until the development of resistance (FIG.4F).
  • mice from the SG-treatment arm were equally distributed into two groups: one to continue SG-treatment and one to receive the combination treatment.
  • Addition of samotolisib fully re-established disease control with no signs of toxicity observed in the combo-treated mice as compared to their SG monotherapy-treated counterparts, as per body weight measurements (FIGs.4F and 8C).
  • Example 6 Imaging TROP-2-expressing DPM models with a novel diagnostic tracer utilizing Zirconium-89; [ 89 Zr]Zr-DFO-SG.
  • PET Immuno-positron emission tomography
  • the utility of a tissue biopsy to assess eligibility for a therapeutic target such as TROP-2 can be limited by intra- and inter-tumoral heterogeneity.
  • Molecular imaging using the TROP-2 antibody can directly assess pharmacodynamic parameters, including temporal biodistribution and tumor specificity of uptake.
  • TROP-2 targeted therapies such as SG.
  • SG was conjugated with the PET radionuclide Zirconium-89 to produce a TROP-2 targeted immunoPET radiotracer.
  • the radiolabeling properties of [ 89 Zr]Zr-DFO- SG, including bioconjugation, radiolabeling, and serum stability assays were deemed satisfactory (FIGs.9A-9C).
  • Serial immunoPET/CT imaging of [ 89 Zr]Zr-DFO-SG in TROP-2 low- and high- expressing models confirmed the potential of [ 89 Zr]Zr-DFO-SG as diagnostic tracer.
  • any listed range can be easily recognized as sufficiently describing and enabling the same range being broken down into at least equal halves, thirds, quarters, fifths, tenths, etc.
  • each range discussed herein can be readily broken down into a lower third, middle third and upper third, etc.
  • all language such as “up to,” “at least,” “greater than,” “less than,” and the like, include the number recited and refer to ranges which can be subsequently broken down into subranges as discussed above.
  • a range includes each individual member.
  • a group having 1-3 cells refers to groups -72- 4932-3602-9469.1 Atty.
  • Colnaghi Biochemical characterization of Trop-2, a cell surface molecule expressed by human carcinomas: formal proof that the monoclonal antibodies T16 and MOv-16 recognize Trop- 2. Hybridoma 11, 539-545 (1992).

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Abstract

The present disclosure relates generally to compositions comprising an antibody-drug conjugate comprising an anti-TROP-2 antibody or antigen binding fragment thereof and methods for using the same to treat diffuse pleural mesothelioma (DPM).

Description

Atty. Dkt. No.115872-3182 TARGETING TROP-2 IN DIFFUSE PLEURAL MESOTHELIOMA CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application claims the benefit of and priority to U.S. Provisional Application No. 63/556,839, filed February 22, 2024, the contents of which are incorporated herein by reference in their entireties. TECHNICAL FIELD [0002] The present disclosure generally relates to antibody-drug conjugates targeting TROP-2 and methods for using the same to treat diffuse pleural mesothelioma (DPM). GOVERNMENT SUPPORT [0003] This invention was made with government support under CA008748 and CA233259-06, awarded by the National Institutes of Health. The government has certain rights in the invention. BACKGROUND [0004] The following description of the background of the present technology is provided simply as an aid in understanding the present technology and is not admitted to describe or constitute prior art to the present technology. [0005] There are only two FDA approved options for patients with diffuse pleural mesotheliomas (DPM) both in the first-line setting. There are no approved therapies in the second line and beyond setting and no targeted therapeutic strategies. There is a striking lack of prospectively validated biomarkers in DPM other than histology. [0006] Accordingly, there is an urgent need for new methods of treating diffuse pleural mesothelioma (DPM). SUMMARY OF THE PRESENT TECHNOLOGY [0007] In one aspect, the present disclosure provides a method for treating diffuse pleural mesothelioma (DPM) in a subject in need thereof comprising administering to the subject an effective amount of an antibody-drug conjugate comprising an anti-TROP-2 antibody or antigen binding fragment thereof. [0008] In one aspect, the present disclosure provides a method for treating diffuse pleural mesothelioma (DPM) in a subject in need thereof comprising administering to the subject an effective amount of an antibody-drug conjugate comprising an anti-TROP-2 -1- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 antibody or antigen binding fragment thereof and an effective amount of an AKT inhibitor. In another aspect, the present disclosure provides a method for re-sensitizing DPM tumors to anti-TROP-2 antibody-drug conjugate therapy comprising administering to the subject an effective amount of an antibody-drug conjugate comprising an anti-TROP-2 antibody or antigen binding fragment thereof and an effective amount of an AKT inhibitor. Additionally or alternatively, in some embodiments, the antibody-drug conjugate and the AKT inhibitor are administered separately, simultaneously, or sequentially. Examples of AKT inhibitors include, but are not limited to, samotolisib, oridonin, capivasertib, ipatasertib, miransertib, afuresertib, uprosertib, BAY1125976, MK-2206, TAS-117, GSK690693, triciribine, and perifosine. In some embodiments, the AKT inhibitor is administered intravenously, intraperitoneally, subcutaneously, intramuscularly, or intratumorally. [0009] In any of the preceding embodiments of the methods disclosed herein, the antibody-drug conjugate comprises one or more of alkylating agents, topoisomerase inhibitors, platinum agents, taxanes, vinca agents, anti-estrogen drugs, aromatase inhibitors, ovarian suppression agents, VEGF/VEGFR inhibitors, EGF/EGFR inhibitors, PARP inhibitors, cytostatic alkaloids, cytotoxic antibiotics, antimetabolites, endocrine/hormonal agents, and bisphosphonate therapy agents. In certain embodiments of the methods disclosed herein, the antibody-drug conjugate comprises a cytotoxic drug selected from the group consisting of an anthracycline, a camptothecin, a tubulin inhibitor, a maytansinoid, a calicheamycin, an auristatin, a nitrogen mustard, an ethylenimine derivative, an alkyl sulfonate, a nitrosourea, a triazene, a folic acid analog, a taxane, a COX-2 inhibitor, a pyrimidine analog, a purine analog, an antibiotic, an enzyme inhibitor, an epipodophyllotoxin, a platinum coordination complex, a vinca alkaloid, a substituted urea, a methyl hydrazine derivative, an adrenocortical suppressant, a hormone antagonist, an antimetabolite, an alkylating agent, an antimitotic, an anti-angiogenic agent, a tyrosine kinase inhibitor, an mTOR inhibitor, a heat shock protein (HSP90) inhibitor, a proteosome inhibitor, an HDAC inhibitor, a topoisomerase inhibitor and a pro-apoptotic agent. [0010] Additionally or alternatively, in some embodiments of the methods described herein, the antibody-drug conjugate comprises cyclophosphamide, fluorouracil (or 5- fluorouracil or 5-FU), methotrexate, edatrexate (10-ethyl-10-deaza-aminopterin), thiotepa, carboplatin, cisplatin, taxanes, paclitaxel, protein-bound paclitaxel, docetaxel, vinorelbine, tamoxifen, raloxifene, toremifene, fulvestrant, gemcitabine, irinotecan, ixabepilone, -2- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 temozolmide, topotecan, vincristine, vinblastine, eribulin, mutamycin, capecitabine, anastrozole, exemestane, letrozole, leuprolide, abarelix, buserlin, goserelin, megestrol acetate, risedronate, pamidronate, ibandronate, alendronate, denosumab, zoledronate, trastuzumab, tykerb, anthracyclines, bevacizumab, Tesirine, afatinib, aplidin, azaribine, axitinib, AVL-101, AVL-291, bendamustine, bleomycin, bortezomib, bosutinib, bryostatin- 1, busulfan, calicheamycin, camptothecin, 10-hydroxy camptothecin, carmustine, celecoxib, chlorambucil, COX-2 inhibitors, SN-38, cladribine, camptothecans, crizotinib, cytarabine, dacarbazine, dasatinib, dinaciclib, dactinomycin, daunorubicin, deruxtecan, DM1, DM3, DM4, doxorubicin, 2-pyrrolinodoxorubicine (2-PDox), a pro-drug form of 2-PDox (pro-2- PDox), cyano-morpholino doxorubicin, doxorubicin glucuronide, endostatin, epirubicin glucuronide, erlotinib, estramustine, epidophyllotoxin, erlotinib, entinostat, estrogen receptor binding agents, etoposide (VP 16), etoposide glucuronide, etoposide phosphate, fmgolimod, floxuridine (FUdR), 3',5'-O-dioleoyl-FudR (FUdR-dO), fludarabine, flutamide, fame syl -protein transferase inhibitors, flavopiridol, fostamatinib, ganetespib, GDC-0834, GS-1101, gefitinib, hydroxyurea, ibrutinib, idarubicin, idelalisib, ifosfamide, imatinib, lapatinib, lenolidamide, leucovorin, LFM-A13, lomustine, mechlorethamine, melphalan, mercaptopurine, 6-mercaptopurine, mitoxantrone, mithramycin, mitomycin, mitotane, monomethylauristatin F (MMAF), monomethylauristatin D (MMAD), monomethylauristatin E (MMAE), navelbine, neratinib, nilotinib, nitrosurea, olaparib, plicomycin, procarbazine, PCI-32765, pentostatin, PSI-341, semustine, SN-38, sorafenib, streptozocin, SU11248, sunitinib, transplatinum, thalidomide, thioguanine, teniposide, uracil mustard, vatalanib, vinca alkaloids, ZD1839 or combinations thereof. [0011] In any of the preceding embodiments of the methods disclosed herein, the anti- TROP-2 antibody or antigen binding fragment comprises the variable heavy (VH) domain and variable light (VL) domain of sacituzumab. In some embodiments of the methods disclosed herein, the VH domain comprises a VH-CDR1 sequence of SEQ ID NO: 5, a VH- CDR2 sequence of SEQ ID NO: 6, and a VH-CDR3 sequence of SEQ ID NO: 7 and the VL domain comprises a VL-CDR1 sequence of SEQ ID NO: 8, a VL-CDR2 sequence of SEQ ID NO: 9, and a VL-CDR3 sequence of SEQ ID NO: 10. Additionally or alternatively, in certain embodiments, (a) the VH domain comprises an amino acid sequence of SEQ ID NO: 3; and (b) the VL domain comprises an amino acid sequence of SEQ ID NO: 1. In some embodiments, the antibody-drug conjugate comprises sacituzumab govitecan. -3- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 [0012] In some embodiments of the methods disclosed herein, the anti-TROP-2 antibody or antigen binding fragment comprises the variable heavy domain and variable light domain of datopotamab. Additionally or alternatively, in certain embodiments of the methods disclosed herein, the VH domain comprises a VH-CDR1 sequence of SEQ ID NO: 15, a VH-CDR2 sequence of SEQ ID NO: 16, and a VH-CDR3 sequence of SEQ ID NO: 17 and the VL domain comprises a VL-CDR1 sequence of SEQ ID NO: 18, a VL-CDR2 sequence of SEQ ID NO: 19, and a VL-CDR3 sequence of SEQ ID NO: 20. In some embodiments of the methods described herein, (a) the VH domain comprises an amino acid sequence of SEQ ID NO: 13; and (b) the VL domain comprises an amino acid sequence of SEQ ID NO: 11. [0013] In any and all embodiments of the methods disclosed herein, the subject has received a prior anti-cancer therapy. Additionally or alternatively, in some embodiments, the subject is diagnosed with or is suffering from recurrent DPM or metastatic DPM. In any and all embodiments of the methods disclosed herein, the antibody-drug conjugate is administered intravenously, intraperitoneally, subcutaneously, intramuscularly, or intratumorally. BRIEF DESCRIPTION OF THE DRAWINGS [0014] FIGs.1A-1C. TROP-2 expression enriched in malignant pleural mesothelioma and associated with worse clinical outcomes. FIG.1A: TROP-2 H-Score, as assessed by #NBP2-49166, Novus Biologicals, in a tissue microarray (TMA) derived from diffuse pleural mesothelioma samples (n=227) and benign pleural samples (n=20) finding significantly enriched expression in DPM (p<0.0001; Mann-Whitney). To the left are representative TROP-2 immunohistochemistry in an example of benign pleura (1) with H-score of 0 and in DPM (2-4) with H-scores of 167, 24 and 11, respectively (original magnifications 200x). FIG.1B: Overall survival (OS) of patients whose samples were included in the DPM TMA noting significantly worse clinical outcomes in patients with TROP- 2 H-Score ≥ 10. FIG.1C: Forrest plot evaluating risk of death using a cox regression analysis noting significantly improved outcomes in patients with low TROP-2 expression (defined as an H-score < 10) and in those with epithelioid histology. [0015] FIGs.2A-2D: TROP-2 expression is associated to the activation of pro- oncogenic pathways in DPM. FIG.2A: Dotplots depicting pathway enrichment analyses in two independent transcriptomic datasets of DPM clinical specimens (TCGA(4) and -4- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 Mesomics(27)), where DGE analyses were run in the top TROP-2-expressing tumors (highest tertile) versus the lowest TROP-2-expressors (lower tertile), followed by pathway enrichment analysis. FIG.2B: Heatmaps showing expression of genes of interest, particularly upregulated genes involved in stemness, EMT/metastasis, and MAPK and AKT signaling in TCGA and Mesomics datasets. FIG.2C: Western blot showing dysregulation of protein markers of interest in isogenic DPM cell lines with ectopic TROP-2 expression dysregulation. Western blotting was reproduced in independent lysates (biological replicates), and a representative image is shown. TROP-2 = TROP-2 overexpression, sgTROP- 2 = TROP-2 KO. FIG.2D: Dotplots showing pathway enrichment analyses after DGE on isogenic TROP-2-overexpressing (TROP-2) or TROP-2-KO (sgTROP-2) cell lines compared to their control counterpart (Control). [0016] FIGs.3A-3F: TROP-2 exerts pro-oncogenic effects in DPM preclinical models. FIG.3A Proliferation curves of isogenic TROP-2-overexpressing (TROP-2) or TROP-2-KO (sgTROP-2) cell lines compared to their control counterpart. In vitro soft agar colony formation (FIG.3B), migration (FIG.3C) and invasion (FIG.3D) of isogenic TROP-2-overexpressing (TROP-2) or TROP-2-KO (sgTROP-2) cell lines compared to their control counterpart. FIG.3E: Imaging of metastasis derived from intracardiac injection of luciferase-expressing DPM cell lines with ectopic overexpression (H28) or KO (H2452) of TROP-2, injected in immunocompromised mice (N=5 per condition), at day 16 post- injection. FIG.3F: Survival curves of mice included in the intracardiac injection experiments in FIG.3E. p-values for FIGS.3A-3D were calculated using the Student’s t- test (unpaired, heterogeneous variances, two-tailed). p-values for FIG.3F were calculated using the Wilcoxon test for survival analysis. p-value legend: *<0.05, **<0.01, ***<0.001. [0017] FIGs.4A-4H: Sacituzumab govitecan shows high efficacy in DPM PDX models. FIG.4A: Heatmap showing TROP-2 expression at the mRNA level (transcriptomic sequencing, in logTPM (transcripts per million ) units) and at the protein level (immunohistochemistry, in H-score, and flow cytometry, in logMFI (mean fluorescence intensity) units). FIG.4B: In vivo treatments with gemcitabine, SG or irinotecan in DPM PDXs including MSK_Lx707 (treatment naive, low TROP-2 expression [control N=10; gemcitabine N=10; SG N=10]), MSK_Lx13 (pre-treated, low expression [control N=6; gemcitabine N=5; SG N=6; irinotecan N=5]), MSK_Lx307 (treatment naive, intermediate expression [control N=10; gemcitabine N=10; SG N=10]) and MSK_Lx606 (pre-treated, high TROP-2 expression [control N=5; gemcitabine N=4; SG N=8; irinotecan -5- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 N=4]). FIG.4C: Dotplots showing pathway enrichment analyses after DGE on SG-treated versus control MSK_Lx307 and MSK_Lx707 DPM PDX tumors. FIG.4D: Western blots showing activation of the PI3K/AKT pathway by assessing phosphorylation of AKT and its downstream effector PRAS40 in SG-treated versus control MSK_Lx307 and MSK_Lx707 DPM PDX tumors. FIG.4E: In vitro synergy assays testing the synergic potential of the combination of SG with the AKT/mTOR inhibitor samotolisib in DPM cell lines with TROP-2 expression, H2452 and TROP-2-ovexpressing MSTO-211H. FIG.4F: In vivo treatment experiment of the MSK_Lx606 DPM PDX (control N=5) with SG until development of SG resistance, moment at which SG-treated mice were divided into SG- monotherapy (N=4) or SG/samotolisib combination (N=4). p-values for FIGs.4B and 4F were calculated using the Student’s t-test (unpaired, heterogeneous variances, two-tailed) comparing the volumes of the control arm or the gemcitabine-treated arm versus the volumes of the SG- treated arm, at experimental endpoint for either the control or the gemcitabine-treated arm. p- value legend: *<0.05, **<0.01, ***<0.001. FIG.4G: ImmunoPET/CT imaging studies with Lx13 (top) and Lx606 (bottom) tumor models from 24 hours post-injection of [89Zr]Zr-DFO-SG (left) to 72 hours for Lx13 (top right), and 120 hours for Lx 606 (bottom left). MIP; Maximum Intensity Projection, % IA/CC; % injected activity per cc. FIG.4H: Corresponding ex vivo biodistribution of main organs Lx13 (blue) and Lx606 (red) at 72 hours and 120 hours post-injection of [89Zr]Zr-DFO- SG. [0018] FIGS.5A-5D: Median overall survival (OS) of patients with diffuse pleural mesotheliomas (DPM) based on TROP-2 expression in a tissue microarray (TMA) at various TROP-2 H-Score cut points: (FIG.5A) 5, (FIG.5B) 10, (FIG.5C) 16.5 [H-score cut point estimated to optimally divide the cohort by survival status], and (FIG.5D) 20. [0019] FIG 6: IVIS imaging of immunocompromised mice right after injection of luciferase-expressing H2452 (control and TROP-2-KO) and H28 (control and TROP-2- overexpressing) to ensure equal administration of cells in all models. [0020] FIGs.7A-7B: FIG.7A: TROP-2 expression in DPM PDXs by IHC. FIG.7B: TROP-2 expression in 25 tested DPM POX models and 3 DPM cells lines by flow cytometry (PE anti-human TACSTD2 [TROP-2, clone NY18], Biolegend; lsotype control [blue]; TROP-2 [red marked with (*)]). [0021] FIGs.8A-8C: FIG.8A: Mouse body weight during in vivo treatments shown in Figure 4B. MSK_Lx707 (treatment naive, low TROP-2 expression [control N=10; -6- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 gemcitabine N=10; Sacituzumab govitecan (SG) N=10]), MSK_Lx13 (pre-treated, low expression [control N=6; gemcitabine N=5; SG N=6; irinotecan N=5]), MSK_Lx307 (treatment naive, intermediate expression [control N=10; gemcitabine N=10; SG N=10]) and MSK_Lx606 (pre- treated, high TROP-2 expression [control N=5; gemcitabine N=4; SG N=8; irinotecan N=4]). FIG.8B: TROP-2 mRNA expression in MSK-Lx307 and MSK- Lx707 DPM PDXs, comparing SG-treated tumor versus the corresponding control tumor. FIG.8C: Mouse body weight during in vivo treatments shown in FIG.4F, after development of resistance to SG (day 160). SG N=4, SG + Samotolisib N=4. p-values for and were calculated using the Student’s t- test (unpaired, heterogeneous variances, two- tailed) [0022] FIGs.9A-9C: Radiochemical characterization and bioconjugation properties of [89Zr]Zr-DFO-SG utilizing instant thin layer chromatography (FIG.9A), serum stability of the radioimmunoconjugate through 144 hours (FIG.9B), and MALDI-TOF for determination of chelator degree of labeling (FIG.9C). [0023] FIG.10 shows the tissue microarray patient demographics. DETAILED DESCRIPTION [0024] It is to be appreciated that certain aspects, modes, embodiments, variations and features of the present methods are described below in various levels of detail in order to provide a substantial understanding of the present technology. [0025] In practicing the present methods, many conventional techniques in molecular biology, protein biochemistry, cell biology, immunology, microbiology and recombinant DNA are used. See, e.g., Sambrook and Russell eds. (2001) Molecular Cloning: A Laboratory Manual, 3rd edition; the series Ausubel et al. eds. (2007) Current Protocols in Molecular Biology; the series Methods in Enzymology (Academic Press, Inc., N.Y.); MacPherson et al. (1991) PCR 1: A Practical Approach (IRL Press at Oxford University Press); MacPherson et al. (1995) PCR 2: A Practical Approach; Harlow and Lane eds. (1999) Antibodies, A Laboratory Manual; Freshney (2005) Culture of Animal Cells: A Manual of Basic Technique, 5th edition; Gait ed. (1984) Oligonucleotide Synthesis; U.S. Patent No.4,683,195; Hames and Higgins eds. (1984) Nucleic Acid Hybridization; Anderson (1999) Nucleic Acid Hybridization; Hames and Higgins eds. (1984) Transcription and Translation; Immobilized Cells and Enzymes (IRL Press (1986)); Perbal (1984) A Practical Guide to Molecular Cloning; Miller and Calos eds. (1987) Gene Transfer Vectors -7- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 for Mammalian Cells (Cold Spring Harbor Laboratory); Makrides ed. (2003) Gene Transfer and Expression in Mammalian Cells; Mayer and Walker eds. (1987) Immunochemical Methods in Cell and Molecular Biology (Academic Press, London); and Herzenberg et al. eds (1996) Weir’s Handbook of Experimental Immunology. Methods to detect and measure levels of polypeptide gene expression products (i.e., gene translation level) are well-known in the art and include the use of polypeptide detection methods such as antibody detection and quantification techniques. (See also, Strachan & Read, Human Molecular Genetics, Second Edition. (John Wiley and Sons, Inc., NY, 1999)). [0026] High expression of TROP-2 (transmembrane protein product of TACSTD2) correlates with tumorigenesis, tumor progression, and inferior outcomes in several solid tumors. Targeting TROP-2 using antibody-drug conjugates (ADCs) such as sacituzumab govitecan (SG) is under active investigation in several malignancies. However, the role of TROP-2 in DPM is poorly defined and ADCs targeting TROP-2 have not been evaluated in DPM. [0027] The present disclosure demonstrates that TROP-2-targeting ADCs are effective in treating aggressive DPM, and that AKT inhibitors can be used to successfully sensitize resistant DPM tumors to TROP-2-targeting ADCs. Definitions [0028] Unless defined otherwise, all technical and scientific terms used herein generally have the same meaning as commonly understood by one of ordinary skill in the art to which this technology belongs. As used in this specification and the appended claims, the singular forms “a”, “an” and “the” include plural referents unless the content clearly dictates otherwise. For example, reference to “a cell” includes a combination of two or more cells, and the like. Generally, the nomenclature used herein and the laboratory procedures in cell culture, molecular genetics, organic chemistry, analytical chemistry and nucleic acid chemistry and hybridization described below are those well-known and commonly employed in the art. [0029] As used herein, the term “about” in reference to a number is generally taken to include numbers that fall within a range of 1%, 5%, or 10% in either direction (greater than or less than) of the number unless otherwise stated or otherwise evident from the context (except where such number would be less than 0% or exceed 100% of a possible value). -8- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 [0030] As used herein, the “administration” of an agent or drug to a subject includes any route of introducing or delivering to a subject a compound to perform its intended function. Administration can be carried out by any suitable route, including but not limited to, orally, intranasally, intrathecally, parenterally (intravenously, intramuscularly, intraperitoneally, or subcutaneously), rectally, intrathecally, intraocularly, intradermally, transmucosally, iontophoretically, or topically. Administration includes self-administration and the administration by another. [0031] As used herein, the term “antibody” collectively refers to immunoglobulins or immunoglobulin-like molecules including by way of example and without limitation, IgA, IgD, IgE, IgG and IgM, combinations and fragments thereof, and similar molecules produced during an immune response in any vertebrate, for example, in mammals such as humans, goats, rabbits and mice, as well as non-mammalian species, such as shark immunoglobulins. Antibodies and antibody fragments can be wholly or partially derived from mammals (e.g., humans, non-human primates, goats, guinea pigs, hamsters, horses, mice, rats, rabbits and sheep) or non-mammalian antibody producing animals (e.g., chickens, ducks, geese, snakes, and urodele amphibians). The antibodies and antibody fragments can be produced in animals or produced outside of animals, such as from yeast or phage (e.g., as a single antibody or antibody fragment or as part of an antibody library). As used herein, “antibodies” (includes intact immunoglobulins) and “antigen binding fragments” specifically bind to a molecule of interest (or a group of highly similar molecules of interest) to the substantial exclusion of binding to other molecules (for example, antibodies and antibody fragments that have a binding constant for the molecule of interest that is at least 103 M-1 greater, at least 104 M-1 greater or at least 105 M-1 greater than a binding constant for other molecules in a biological sample). The term “antibody” also includes genetically engineered forms such as chimeric antibodies (for example, humanized murine antibodies), heteroconjugate antibodies (such as, bispecific antibodies). See also, Pierce Catalog and Handbook, 1994-1995 (Pierce Chemical Co., Rockford, Ill.); Kuby, J., Immunology, 3rd Ed., W.H. Freeman & Co., New York, 1997. Antibodies may comprise whole native antibodies, monoclonal antibodies, human antibodies, humanized antibodies, camelised antibodies, multispecific antibodies, bispecific antibodies, chimeric antibodies, Fab, Fab', single chain V region fragments (scFv), single domain antibodies (e.g., nanobodies and single domain camelid antibodies), VNAR fragments, Bi-specific T- cell engager (BiTE) antibodies, minibodies, disulfide-linked Fvs (sdFv), and anti-idiotypic -9- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 (anti-Id) antibodies, intrabodies, fusion polypeptides, unconventional antibodies and antigen binding fragments of any of the above. [0032] More particularly, antibody refers to a polypeptide ligand comprising at least a light chain immunoglobulin variable region or heavy chain immunoglobulin variable region which specifically recognizes and binds an epitope of an antigen. Antibodies are composed of a heavy and a light chain each of which has a variable region, termed the variable heavy (VH) region and the variable light (VL) region. The heavy chain constant (CH) region is comprised of three domains, CH1, CH2, and CH3. Together, the VH region and the VL region are responsible for binding the antigen recognized by the antibody. Typically, an immunoglobulin has heavy (H) chains and light (L) chains interconnected by disulfide bonds. There are two types of light chain, lambda (λ) and kappa (^). There are five main heavy chain classes (or isotypes) which determine the functional activity of an antibody molecule: IgM, IgD, IgG, IgA and IgE. Immunoglobulin molecules can be of any type (e.g., IgG, IgE, IgM, IgD, IgA, and IgY), class (e.g., IgGl, IgG2, IgG3, IgG4, IgAl, and IgA2), or subclass. Each heavy and light chain contains a constant region and a variable region, (the regions are also known as “domains”). In combination, the heavy and the light chain variable regions specifically bind the antigen. Light and heavy chain variable regions contain a “framework” region interrupted by three hypervariable regions, also called “complementarity-determining regions” or “CDRs”. The extent of the framework region and CDRs have been defined (see, Kabat et al., Sequences of Proteins of Immunological Interest, U.S. Department of Health and Human Services, 1991, which is hereby incorporated by reference). The Kabat database is now maintained online. The sequences of the framework regions (FR) of different light or heavy chains are relatively conserved within a species. Each VH and VL is composed of three CDRs and four FRs arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. The framework region of an antibody, that is the combined framework regions of the constituent light and heavy chains, largely adopt a β-sheet conformation and the CDRs form loops which connect, and in some cases form part of, the β-sheet structure. Thus, framework regions act to form a scaffold that provides for positioning the CDRs in correct orientation by inter-chain, non-covalent interactions. [0033] The CDRs are primarily responsible for binding to an epitope of an antigen. The CDRs of each chain are typically referred to as CDR1, CDR2, and CDR3 and are also typically identified by the chain in which the particular CDR is located. Thus, a VH CDR3 -10- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 is located in the variable domain of the heavy chain of the antibody in which it is found, whereas a VL CDR1 is the CDR1 from the variable domain of the light chain of the antibody in which it is found. An antibody that binds TROP-2 protein will have a specific VH region and VL region sequence, and thus specific CDR sequences. Antibodies with different specificities (i.e. different combining sites for different antigens) have different CDRs. [0034] Although it is the CDRs that vary from antibody to antibody, only a limited number of amino acid positions within the CDRs are directly involved in antigen binding. These positions within the CDRs are called specificity determining residues (SDRs). “Immunoglobulin-related compositions” as used herein, refers to antibodies (including monoclonal antibodies, polyclonal antibodies, humanized antibodies, chimeric antibodies, recombinant antibodies, multi-specific antibodies, bispecific antibodies, etc.,) as well as antibody fragments. An antibody or antigen binding fragment thereof specifically binds to an antigen. [0035] As used herein, the term “antibody-related polypeptide” means antigen-binding antibody fragments, including single-chain antibodies, that can comprise the variable region(s) alone, or in combination, with all or part of the following polypeptide elements: hinge region, CH1, CH2, and CH3 domains of an antibody molecule. Also included in the technology are any combinations of variable region(s) and hinge region, CH1, CH2, and CH3 domains. Antibody-related molecules useful in the present methods, e.g., but are not limited to, Fab, Fab′ and F(ab′)2, Fd, single-chain Fvs (scFv), single-chain antibodies, disulfide-linked Fvs (sdFv) and fragments comprising either a VL or VH domain. Examples include: (i) a Fab fragment, a monovalent fragment consisting of the VL, VH, CL and CH1 domains; (ii) a F(ab′)2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; (iii) a Fd fragment consisting of the VH and CH1 domains; (iv) a Fv fragment consisting of the VL and VH domains of a single arm of an antibody, (v) a dAb fragment (Ward et al., Nature 341: 544-546, 1989), which consists of a VH domain; and (vi) an isolated complementarity determining region (CDR). As such “antibody fragments” or “antigen binding fragments” can comprise a portion of a full length antibody, generally the antigen binding or variable region thereof. Examples of antibody fragments or antigen binding fragments include Fab, Fab', F(ab')2, and Fv fragments; diabodies; linear antibodies; single-chain antibody molecules; and multi-specific antibodies formed from antibody fragments. -11- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 [0036] As used herein, the term “diabodies” refers to small antibody fragments with two antigen-binding sites, which fragments comprise a heavy-chain variable domain (VH) connected to a light-chain variable domain (VL) in the same polypeptide chain (VH VL). By using a linker that is too short to allow pairing between the two domains on the same chain, the domains are forced to pair with the complementary domains of another chain and create two antigen binding sites. Diabodies are described more fully in, e.g., EP 404,097; WO 93/11161; and Hollinger et al., Proc. Natl. Acad. Sci. USA, 90: 6444-6448 (1993). [0037] As used herein, the terms “single-chain antibodies” or “single-chain Fv (scFv)” refer to an antibody fusion molecule of the two domains of the Fv fragment, VL and VH. Single-chain antibody molecules may comprise a polymer with a number of individual molecules, for example, dimer, trimer or other polymers. Furthermore, although the two domains of the Fv fragment, VL and VH, are coded for by separate genes, they can be joined, using recombinant methods, by a synthetic linker that enables them to be made as a single protein chain in which the VL and VH regions pair to form monovalent molecules (known as single-chain Fv (scFv)). Bird et al. (1988) Science 242:423-426 and Huston et al. (1988) Proc. Natl. Acad Sci. USA 85:5879-5883. Such single-chain antibodies can be prepared by recombinant techniques or enzymatic or chemical cleavage of intact antibodies. [0038] Any of the above-noted antibody fragments are obtained using conventional techniques known to those of skill in the art, and the fragments are screened for binding specificity and neutralization activity in the same manner as are intact antibodies. [0039] The term “antigen binding fragment” refers to a fragment of the whole immunoglobulin structure which possesses a part of a polypeptide responsible for binding to antigen. Examples of the antigen binding fragment useful in the present technology include scFv, (scFv)2, scFvFc, Fab, Fab′ and F(ab′)2, but are not limited thereto. Any of the above- noted antibody fragments are obtained using conventional techniques known to those of skill in the art, and the fragments are screened for binding specificity and neutralization activity in the same manner as are intact antibodies. [0040] As used herein, an “antigen” refers to a molecule to which an antibody (or antigen binding fragment thereof) can selectively bind. The target antigen may be a protein, carbohydrate, nucleic acid, lipid, hapten, or other naturally occurring or synthetic compound. In some embodiments, the target antigen may be a polypeptide (e.g., a TROP-2 -12- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 polypeptide). An antigen may also be administered to an animal to generate an immune response in the animal. [0041] By “binding affinity” is meant the strength of the total noncovalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen or antigenic peptide). The affinity of a molecule X for its partner Y can generally be represented by the dissociation constant (KD). Affinity can be measured by standard methods known in the art, including those described herein. A low-affinity complex contains an antibody that generally tends to dissociate readily from the antigen, whereas a high-affinity complex contains an antibody that generally tends to remain bound to the antigen for a longer duration. [0042] As used herein, the term “biological sample” means sample material derived from living cells. Biological samples may include tissues, cells, protein or membrane extracts of cells, and biological fluids (e.g., ascites fluid or cerebrospinal fluid (CSF)) isolated from a subject, as well as tissues, cells and fluids present within a subject. Biological samples of the present technology include, but are not limited to, samples taken from breast tissue, renal tissue, the uterine cervix, the endometrium, the head or neck, the gallbladder, parotid tissue, the prostate, the brain, the pituitary gland, kidney tissue, muscle, the esophagus, the stomach, the small intestine, the colon, the liver, the spleen, the pancreas, thyroid tissue, heart tissue, lung tissue, the bladder, adipose tissue, lymph node tissue, the uterus, ovarian tissue, adrenal tissue, testis tissue, the tonsils, thymus, blood, hair, buccal, skin, serum, plasma, CSF, semen, prostate fluid, seminal fluid, urine, feces, sweat, saliva, sputum, mucus, bone marrow, lymph, and tears. Biological samples can also be obtained from biopsies of internal organs or from cancers. Biological samples can be obtained from subjects for diagnosis or research or can be obtained from non-diseased individuals, as controls or for basic research. Samples may be obtained by standard methods including, e.g., venous puncture and surgical biopsy. In certain embodiments, the biological sample is a tissue sample obtained by needle biopsy. [0043] As used herein, the term “CDR-grafted antibody” means an antibody in which at least one CDR of an “acceptor” antibody is replaced by a CDR “graft” from a “donor” antibody possessing a desirable antigen specificity. [0044] As used herein, the term “chimeric antibody” means an antibody in which the Fc constant region of a monoclonal antibody from one species (e.g., a mouse Fc constant -13- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 region) is replaced, using recombinant DNA techniques, with an Fc constant region from an antibody of another species (e.g., a human Fc constant region). See generally, Robinson et al., PCT/US86/02269; Akira et al., European Patent Application 184,187; Taniguchi, European Patent Application 171,496; Morrison et al., European Patent Application 173,494; Neuberger et al., WO 86/01533; Cabilly et al. U.S. Patent No.4,816,567; Cabilly et al., European Patent Application 0125,023; Better et al., Science 240: 1041-1043, 1988; Liu et al., Proc. Natl. Acad. Sci. USA 84: 3439-3443, 1987; Liu et al., J. Immunol 139: 3521-3526, 1987; Sun et al., Proc. Natl. Acad. Sci. USA 84: 214-218, 1987; Nishimura et al., Cancer Res 47: 999-1005, 1987; Wood et al., Nature 314: 446-449, 1885; and Shaw et al., J. Natl. Cancer Inst.80: 1553-1559, 1988. [0045] As used herein, the term “conjugated” refers to the association of two molecules by any method known to those in the art. Suitable types of associations include chemical bonds and physical bonds. Chemical bonds include, for example, covalent bonds and coordinate bonds. Physical bonds include, for instance, hydrogen bonds, dipolar interactions, van der Waal forces, electrostatic interactions, hydrophobic interactions and aromatic stacking. [0046] As used herein, the term “consensus FR” means a framework (FR) antibody region in a consensus immunoglobulin sequence. The FR regions of an antibody do not contact the antigen. [0047] As used herein, a "control" is an alternative sample used in an experiment for comparison purpose. A control can be "positive" or "negative." For example, where the purpose of the experiment is to determine a correlation of the efficacy of a therapeutic agent for the treatment for a particular type of disease, a positive control (a compound or composition known to exhibit the desired therapeutic effect) and a negative control (a subject or a sample that does not receive the therapy or receives a placebo) are typically employed. [0048] As used herein, the term “effective amount” refers to a quantity sufficient to achieve a desired therapeutic and/or prophylactic effect, e.g., an amount which results in the prevention of, or a decrease in a disease or condition described herein or one or more signs or symptoms associated with a disease or condition described herein. In the context of therapeutic or prophylactic applications, the amount of a composition administered to the subject will vary depending on the composition, the degree, type, and severity of the disease -14- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 and on the characteristics of the individual, such as general health, age, sex, body weight and tolerance to drugs. The skilled artisan will be able to determine appropriate dosages depending on these and other factors. The compositions can also be administered in combination with one or more additional therapeutic compounds. In the methods described herein, the therapeutic compositions may be administered to a subject having one or more signs or symptoms of a disease or condition described herein. As used herein, a "therapeutically effective amount" of a composition refers to composition levels in which the physiological effects of a disease or condition are ameliorated or eliminated. A therapeutically effective amount can be given in one or more administrations. [0049] As used herein, the term “epitope” means a protein determinant capable of specific binding to an antibody. Epitopes usually consist of chemically active surface groupings of molecules such as amino acids or sugar side chains and usually have specific three dimensional structural characteristics, as well as specific charge characteristics. Conformational and non-conformational epitopes are distinguished in that the binding to the former but not the latter is lost in the presence of denaturing solvents. In some embodiments, an “epitope” of the TROP-2 protein is a region of the protein to which the anti-TROP-2 antibodies of the present technology specifically bind. In some embodiments, the epitope is a conformational epitope or a non-conformational epitope. To screen for anti- TROP-2 antibodies which bind to an epitope, a routine cross-blocking assay such as that described in Antibodies, A Laboratory Manual, Cold Spring Harbor Laboratory, Ed Harlow and David Lane (1988), can be performed. This assay can be used to determine if an anti- TROP-2 antibody binds the same site or epitope as an anti-TROP-2 antibody of the present technology. Alternatively, or additionally, epitope mapping can be performed by methods known in the art. For example, the antibody sequence can be mutagenized such as by alanine scanning, to identify contact residues. In a different method, peptides corresponding to different regions of TROP-2 protein can be used in competition assays with the test antibodies or with a test antibody and an antibody with a characterized or known epitope. [0050] As used herein, “expression” includes one or more of the following: transcription of the gene into precursor mRNA; splicing and other processing of the precursor mRNA to produce mature mRNA; mRNA stability; translation of the mature mRNA into protein (including codon usage and tRNA availability); and glycosylation and/or other modifications of the translation product, if required for proper expression and function. -15- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 [0051] As used herein, the term “gene” means a segment of DNA that contains all the information for the regulated biosynthesis of an RNA product, including promoters, exons, introns, and other untranslated regions that control expression. [0052] “Homology” or “identity” or “similarity” refers to sequence similarity between two peptides or between two nucleic acid molecules. Homology can be determined by comparing a position in each sequence which may be aligned for purposes of comparison. When a position in the compared sequence is occupied by the same base or amino acid, then the molecules are homologous at that position. A degree of homology between sequences is a function of the number of matching or homologous positions shared by the sequences. A polynucleotide or polynucleotide region (or a polypeptide or polypeptide region) has a certain percentage (for example, at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99%) of “sequence identity” to another sequence means that, when aligned, that percentage of bases (or amino acids) are the same in comparing the two sequences. This alignment and the percent homology or sequence identity can be determined using software programs known in the art. In some embodiments, default parameters are used for alignment. One alignment program is BLAST, using default parameters. In particular, programs are BLASTN and BLASTP, using the following default parameters: Genetic code=standard; filter=none; strand=both; cutoff=60; expect=10; Matrix=BLOSUM62; Descriptions=50 sequences; sort by ═HIGH SCORE; Databases=non-redundant, GenBank+EMBL+DDBJ+PDB+GenBank CDS translations+SwissProtein+SPupdate+PIR. Details of these programs can be found at the National Center for Biotechnology Information. Biologically equivalent polynucleotides are those having the specified percent homology and encoding a polypeptide having the same or similar biological activity. Two sequences are deemed “unrelated” or “non-homologous” if they share less than 40% identity, or less than 25% identity, with each other. [0053] As used herein, “humanized” forms of non-human (e.g., murine) antibodies are chimeric antibodies which contain minimal sequence derived from non-human immunoglobulin. For the most part, humanized antibodies are human immunoglobulins in which hypervariable region residues of the recipient are replaced by hypervariable region residues from a non-human species (donor antibody) such as mouse, rat, rabbit or nonhuman primate having the desired specificity, affinity, and capacity. In some embodiments, Fv framework region (FR) residues of the human immunoglobulin are replaced by corresponding non-human residues. Furthermore, humanized antibodies may -16- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 comprise residues which are not found in the recipient antibody or in the donor antibody. These modifications are made to further refine antibody performance such as binding affinity. Generally, the humanized antibody will comprise substantially all of at least one, and typically two, variable domains (e.g., Fab, Fab′, F(ab′)2, or Fv), in which all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin and all or substantially all of the FR regions are those of a human immunoglobulin consensus FR sequence although the FR regions may include one or more amino acid substitutions that improve binding affinity. The number of these amino acid substitutions in the FR are typically no more than 6 in the H chain, and in the L chain, no more than 3. The humanized antibody optionally may also comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. For further details, see Jones et al., Nature 321:522-525 (1986); Reichmann et al., Nature 332:323-329 (1988); and Presta, Curr. Op. Struct. Biol.2:593-596 (1992). See e.g., Ahmed & Cheung, FEBS Letters 588(2):288-297 (2014). [0054] As used herein, the term “hypervariable region” refers to the amino acid residues of an antibody which are responsible for antigen-binding. The hypervariable region generally comprises amino acid residues from a “complementarity determining region” or “CDR” (e.g., around about residues 24-34 (L1), 50-56 (L2) and 89-97 (L3) in the VL, and around about 31-35B (H1), 50-65 (H2) and 95-102 (H3) in the VH (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD. (1991)) and/or those residues from a “hypervariable loop” (e.g., residues 26-32 (L1), 50-52 (L2) and 91-96 (L3) in the VL, and 26-32 (H1), 52A-55 (H2) and 96-101 (H3) in the VH (Chothia and Lesk J. Mol. Biol.196:901-917 (1987)). [0055] As used herein, the terms “identical” or percent “identity”, when used in the context of two or more nucleic acids or polypeptide sequences, refer to two or more sequences or subsequences that are the same or have a specified percentage of amino acid residues or nucleotides that are the same (i.e., about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or higher identity over a specified region (e.g., nucleotide sequence encoding an antibody described herein or amino acid sequence of an antibody described herein)), when compared and aligned for maximum correspondence over a comparison window or designated region as measured using a BLAST or BLAST 2.0 sequence comparison algorithms with default parameters described below, or by manual alignment and visual inspection (e.g., NCBI web site). Such sequences -17- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 are then said to be “substantially identical.” This term also refers to, or can be applied to, the complement of a test sequence. The term also includes sequences that have deletions and/or additions, as well as those that have substitutions. In some embodiments, identity exists over a region that is at least about 25 amino acids or nucleotides in length, or 50-100 amino acids or nucleotides in length. [0056] As used herein, the term “intact antibody” or “intact immunoglobulin” means an antibody that has at least two heavy (H) chain polypeptides and two light (L) chain polypeptides interconnected by disulfide bonds. Each heavy chain is comprised of a heavy chain variable region (abbreviated herein as HCVR or VH) and a heavy chain constant region. The heavy chain constant region is comprised of three domains, CH1, CH2 and CH3. Each light chain is comprised of a light chain variable region (abbreviated herein as LCVR or VL) and a light chain constant region. The light chain constant region is comprised of one domain, CL. The VH and VL regions can be further subdivided into regions of hypervariability, termed complementarity determining regions (CDR), interspersed with regions that are more conserved, termed framework regions (FR). Each VH and VL is composed of three CDRs and four FRs, arranged from amino-terminus to carboxyl-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains contain a binding domain that interacts with an antigen. The constant regions of the antibodies can mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (Clq) of the classical complement system. [0057] The term “monoclonal antibody” as used herein refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations that may be present in minor amounts. For example, a monoclonal antibody can be an antibody that is derived from a single clone, including any eukaryotic, prokaryotic, or phage clone, and not the method by which it is produced. A monoclonal antibody composition displays a single binding specificity and affinity for a particular epitope. Monoclonal antibodies are highly specific, being directed against a single antigenic site. Furthermore, in contrast to conventional (polyclonal) antibody preparations which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen. The modifier “monoclonal” indicates the character of the antibody as being obtained from a substantially homogeneous -18- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 population of antibodies, and is not to be construed as requiring production of the antibody by any particular method. Monoclonal antibodies can be prepared using a wide variety of techniques known in the art including, e.g., but not limited to, hybridoma, recombinant, and phage display technologies. For example, the monoclonal antibodies to be used in accordance with the present methods may be made by the hybridoma method first described by Kohler et al., Nature 256:495 (1975), or may be made by recombinant DNA methods (See, e.g., U.S. Patent No.4,816,567). The “monoclonal antibodies” may also be isolated from phage antibody libraries using the techniques described in Clackson et al., Nature 352:624-628 (1991) and Marks et al., J. Mol. Biol.222:581-597 (1991), for example. [0058] As used herein, the term “pharmaceutically-acceptable carrier” is intended to include any and all solvents, dispersion media, coatings, antibacterial and antifungal compounds, isotonic and absorption delaying compounds, and the like, compatible with pharmaceutical administration. Pharmaceutically-acceptable carriers and their formulations are known to one skilled in the art and are described, for example, in Remington's Pharmaceutical Sciences (20th edition, ed. A. Gennaro, 2000, Lippincott, Williams & Wilkins, Philadelphia, Pa.). [0059] As used herein, the term “polyclonal antibody” means a preparation of antibodies derived from at least two (2) different antibody-producing cell lines. The use of this term includes preparations of at least two (2) antibodies that contain antibodies that specifically bind to different epitopes or regions of an antigen. [0060] As used herein, the term “polynucleotide” or “nucleic acid” means any RNA or DNA, which may be unmodified or modified RNA or DNA. Polynucleotides include, without limitation, single- and double-stranded DNA, DNA that is a mixture of single- and double-stranded regions, single- and double-stranded RNA, RNA that is mixture of single- and double-stranded regions, and hybrid molecules comprising DNA and RNA that may be single-stranded or, more typically, double-stranded or a mixture of single- and double- stranded regions. In addition, polynucleotide refers to triple-stranded regions comprising RNA or DNA or both RNA and DNA. The term polynucleotide also includes DNAs or RNAs containing one or more modified bases and DNAs or RNAs with backbones modified for stability or for other reasons. [0061] As used herein, the terms “polypeptide,” “peptide” and “protein” are used interchangeably herein to mean a polymer comprising two or more amino acids joined to -19- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 each other by peptide bonds or modified peptide bonds, i.e., peptide isosteres. Polypeptide refers to both short chains, commonly referred to as peptides, glycopeptides or oligomers, and to longer chains, generally referred to as proteins. Polypeptides may contain amino acids other than the 20 gene-encoded amino acids. Polypeptides include amino acid sequences modified either by natural processes, such as post-translational processing, or by chemical modification techniques that are well known in the art. Such modifications are well described in basic texts and in more detailed monographs, as well as in a voluminous research literature. [0062] As used herein, “prevention” or “preventing” of a disorder or condition refers to a compound that, in a statistical sample, reduces the occurrence of the disorder or condition in the treated sample relative to an untreated control sample, or delays the onset of one or more symptoms of the disorder or condition relative to the untreated control sample. [0063] As used herein, the term “recombinant” when used with reference, e.g., to a cell, or nucleic acid, protein, or vector, indicates that the cell, nucleic acid, protein or vector, has been modified by the introduction of a heterologous nucleic acid or protein or the alteration of a native nucleic acid or protein, or that the material is derived from a cell so modified. Thus, for example, recombinant cells express genes that are not found within the native (non-recombinant) form of the cell or express native genes that are otherwise abnormally expressed, under expressed or not expressed at all. [0064] As used herein, the term “separate” therapeutic use refers to an administration of at least two active ingredients at the same time or at substantially the same time by different routes. [0065] As used herein, the term “sequential” therapeutic use refers to administration of at least two active ingredients at different times, the administration route being identical or different. More particularly, sequential use refers to the whole administration of one of the active ingredients before administration of the other or others commences. It is thus possible to administer one of the active ingredients over several minutes, hours, or days before administering the other active ingredient or ingredients. There is no simultaneous treatment in this case. [0066] As used herein, “specifically binds” refers to a molecule (e.g., an antibody or antigen binding fragment thereof) which recognizes and binds another molecule (e.g., an antigen), but that does not substantially recognize and bind other molecules. The terms -20- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 “specific binding,” “specifically binds to,” or is “specific for” a particular molecule (e.g., a polypeptide, or an epitope on a polypeptide), as used herein, can be exhibited, for example, by a molecule having a KD for the molecule to which it binds to of about 10−4 M, 10−5 M, 10−6 M, 10−7 M, 10−8 M, 10−9 M, 10−10 M, 10−11 M, or 10−12 M. The term “specifically binds” may also refer to binding where a molecule (e.g., an antibody or antigen binding fragment thereof) binds to a particular polypeptide (e.g., a TROP-2 polypeptide), or an epitope on a particular polypeptide, without substantially binding to any other polypeptide, or polypeptide epitope. [0067] As used herein, the term “simultaneous” therapeutic use refers to the administration of at least two active ingredients by the same route and at the same time or at substantially the same time. [0068] As used herein, the terms “subject”, “patient”, or “individual” can be an individual organism, a vertebrate, a mammal, or a human. In some embodiments, the subject, patient, or individual is a human. [0069] As used herein, the term “therapeutic agent” is intended to mean a compound that, when present in an effective amount, produces a desired therapeutic effect on a subject in need thereof. [0070] “Treating” or “treatment” as used herein covers the treatment of a disease or disorder described herein, in a subject, such as a human, and includes: (i) inhibiting a disease or disorder, i.e., arresting its development; (ii) relieving a disease or disorder, i.e., causing regression of the disorder; (iii) slowing progression of the disorder; and/or (iv) inhibiting, relieving, or slowing progression of one or more symptoms of the disease or disorder. Therapeutic effects of treatment include, without limitation, inhibiting recurrence of disease, alleviation of symptoms, diminishment of any direct or indirect pathological consequences of the disease, preventing metastases, decreasing the rate of disease progression, amelioration or palliation of the disease state, and remission or improved prognosis. By “treating a cancer” is meant that the symptoms associated with the cancer are, e.g., alleviated, reduced, cured, or placed in a state of remission. [0071] It is also to be appreciated that the various modes of treatment of disorders as described herein are intended to mean “substantial,” which includes total but also less than total treatment, and wherein some biologically or medically relevant result is achieved. The -21- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 treatment may be a continuous prolonged treatment for a chronic disease or a single, or few time administrations for the treatment of an acute condition. Immunoglobulin-related Compositions of the Present Technology [0072] The present technology describes methods and compositions for the generation and use of anti-TROP-2 immunoglobulin-related compositions (e.g., anti-TROP-2 antibodies or antigen binding fragments thereof). The anti-TROP-2 immunoglobulin- related compositions of the present disclosure may be useful in the treatment of anti-cancer drug-related skin rashes. Anti-TROP-2 immunoglobulin-related compositions within the scope of the present technology include, e.g., but are not limited to, monoclonal, chimeric, humanized, bispecific antibodies and diabodies that specifically bind the target polypeptide, a homolog, derivative or a fragment thereof. The present disclosure also provides antigen binding fragments of any of the anti-TROP-2 antibodies disclosed herein, wherein the antigen binding fragment is selected from the group consisting of Fab, F(ab)'2, Fab’, scFv, and Fv. [0073] In some aspects, the anti-TROP2 can comprise a commercially available antibody or antibody having the six CDRs of a commercially available antibody selected from the group consisting of LS-C126418, LS-C178765, LS-C126416, LS-C126417 (LifeSpan BioSciences, Inc., Seattle, Wash.); 10428-MM01, 10428-MM02, 10428-R001, 10428- R030 (Sino Biological Inc., Beijing, China); MR54 (eBioscience, San Diego, Calif.); sc- 376181, sc-376746, Santa Cruz Biotechnology (Santa Cruz, Calif.); MM0588- 49D6, (Novus Biologicals, Littleton, Colo.); ab79976, and ab89928 (ABCAM®, Cambridge, Mass.). [0074] In some aspects, the anti-Trop-2 antibody can be selected from sacituzumab or another known anti-Trop antibody such as any of the following: U.S. Publ. No. 2013/0089872 discloses anti-TROP2 antibodies K5-70 (Accession No. FERM BP- 11251), K5-107 (Accession No. FERM BP- 11252), K5-116-2-1 (Accession No. FERM BP-11253), T6-16 (Accession No. FERM BP- 11346), and T5-86 (Accession No. FERM BP-11254), deposited with the International Patent Organism Depositary, Tsukuba, Japan. U.S. Pat. No. 5,840,854 disclosed the anti-TROP2 monoclonal antibody BRI 10 (ATCC No. HB11698). U.S. Pat. No.7,420,040 disclosed an anti-TROP2 antibody produced by hybridoma cell line AR47A6.4.2, deposited with the ID AC (International Depository Authority of Canada, Winnipeg, Canada) as accession number 141205-05. U.S. Pat. No.7,420,041 disclosed an anti-TROP2 antibody produced by hybridoma cell line AR52A301.5, deposited with the -22- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 IDAC as accession number 141205-03. U.S. Publ. No.2013/0122020 disclosed anti-Trop-2 antibodies 3E9, 6G11, 7E6, 15E2, 18B1. Hybridomas encoding a representative antibody were deposited with the American Type Culture Collection (ATCC), Accession Nos. PTA- 12871 and PTA-12872. U.S. Pat. No.8,715,662 discloses anti-Trop-2 antibodies produced by hybridomas deposited at the AID-ICLC (Genoa, Italy) with deposit numbers PD 08019, PD 08020 and PD 08021. U.S. Patent Application Publ. No.20120237518 discloses anti- TROP 2 antibodies 77220, KM4097 and KM4590. U.S. Pat. No.8,309,094 (Wyeth) discloses antibodies Al and A3, identified by sequence listing. [0075] In some aspects, the anti-TROP-2 antibody is sacituzumab, which is also known as the humanized monoclonal antibody hRS7 (e.g., U.S. Pat. No.7,238,785, incorporated herein by reference in its entirety). The sacituzumab antibody was generated using a murine IgGl raised against a crude membrane preparation of a human primary squamous cell lung carcinoma. (Stein et al., Cancer Res.50: 1330, 1990). [0076] Sacituzumab and antigen-binding fragments thereof for use in the methods provided herein comprises a heavy chain and a light chain or a heavy chain variable region and a light chain variable region. Those of ordinary skill in the art would easily be able to identify Chothia-defined, Abm-defined or other CDRs. SEQ ID NO: 1 Sacituzumab VL domain DIQLTQSPSSLSASVGDRVSITCKASQDVSIAVAWYQQKPGKAPK LLIYSASYRYTGVPDRFSGSGSGTDFTLTISSLQPEDFAVYYCQQH YITPLTFGAGTKVEIK SEQ ID NO: 2 Sacituzumab Light Chain DIQLTQSPSSLSASVGDRVSITCKASQDVSIAVAWYQQKPGKAPKLLIY SASYRYTGVPDRFSGSGSGTDFTLTISSLQPEDFAVYYCQQHYITPLTF GAGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREA KVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK HKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 3 Sacituzumab VH domain QVQLQQSGSELKKPGASVKVSCKASGYTFTNYGMNWVKQAPG QGLKWMGWINTYTGEPTYTDDFKGRFAFSLDTSVSTAYLQISSL KADDTAVYFCARGGFGSSYWYFDVWGQGSLVTVSS -23- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 SEQ ID NO: 4 Sacituzumab Heavy Chain QVQLQQSGSELKKPGASVKVSCKASGYTFTNYGMNWVKQAPGQGL KWMGWINTYTGEPTYTDDFKGRFAFSLDTSVSTAYLQISSLKADDTA VYFCARGGFGSSYWYFDVWGQGSLVTVSSASTKGPSVFPLAPSSKST SGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL YSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTH TCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHED PEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD GSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP GK SEQ ID NO: 5 Sacituzumab VH CDR1 NYGMN SEQ ID NO: 6 Sacituzumab VH CDR2 WINTYTGEPTYTDDFKG SEQ ID NO: 7 Sacituzumab VH CDR3 GGFGSSYWYFDV SEQ ID NO: 8 Sacituzumab VL CDR1 KASQDVSIAVA SEQ ID NO: 9 Sacituzumab VL CDR2 SASYRYT SEQ ID NO: Sacituzumab VL CDR3 10 QQHYITPLT VH and VL domains are italicized, VH and VL CDR regions are underlined [0077] Datopotamab deruxtecan has shown clinical efficacy in multiple tumor types, including lung cancer and breast cancer. The TROP2-targeting antibody Datopotamab is described in detail in WO2020/240467. -24- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 [0078] Datopotamab and antigen-binding fragments thereof for use in the methods provided herein comprises a heavy chain and a light chain or a heavy chain variable region and a light chain variable region. Those of ordinary skill in the art would easily be able to identify Chothia-defined, Abm-defined or other CDRs. SEQ ID Datopotamab VL domain NO: 11 DIQMTQSPSSLSASVGDRVTITCKASQDVSTAVAWYQQKPGKAPKL LIYSASYRYTGVPSRFSGSGSGTDFTLTISSLQPEDFAVYYCQQHYITP LTFGQGTKLEIK SEQ ID Datopotamab Light Chain NO: 12 DIQMTQSPSSLSASVGDRVTITCKASQDVSTAVAWYQQKPGKAPKLLIYS ASYRYTGVPSRFSGSGSGTDFTLTISSLQPEDFAVYYCQQHYITPLTFGQG TKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQW KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYAC EVTHQGLSSPVTKSFNRGEC SEQ ID Datopotamab VH domain NO: 13 QVQLVQSGAEVKKPGASVKVSCKASGYTFTTAGMQWVRQAPGQG LEWMGWINTHSGVPKYAEDFKGRVTISADTSTSTAYLQLSSLKSEDT AVYYCARSGFGSSYWYFDVWGQGTLVTVSS SEQ ID Datopotamab Heavy Chain NO: 14 QVQLVQSGAEVKKPGASVKVSCKASGYTFTTAGMQWVRQAPGQGLEW MGWINTHSGVPKYAEDFKGRVTISADTSTSTAYLQLSSLKSEDTAVYYCAR SGFGSSYWYFDVWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAAL GCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVP SSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLG GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQG NVFSCSVMHEALHNHYTQKSLSLSPGK -25- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 SEQ ID Datopotamab VH CDR1 NO: 15 TAGMQ SEQ ID Datopotamab VH CDR2 NO: 16 WINTHSGVPKYAEDFK SEQ ID Datopotamab VH CDR3 NO: 17 SGFGSSYWYFDV SEQ ID Datopotamab VL CDR1 NO: 18 KASQDVSTAVA SEQ ID Datopotamab VL CDR2 NO: 19 SASYRYT SEQ ID Datopotamab VL CDR3 NO: 20 QQHYITPLT [0079] In one aspect, the present disclosure provides a TROP-2 antibody or antigen binding fragment thereof comprising a heavy chain immunoglobulin variable domain (VH) and a light chain immunoglobulin variable domain (VL), wherein the VH comprises a VH- CDR1 sequence of SEQ ID NO: 5, a VH-CDR2 sequence of SEQ ID NO: 6, and a VH- CDR3 sequence of SEQ ID NO: 7 and the VL comprises a VL-CDR1 sequence of SEQ ID NO: 8, a VL-CDR2 sequence of SEQ ID NO: 9, and a VL-CDR3 sequence of SEQ ID NO: 10. In one aspect, the present disclosure provides a TROP-2 antibody or antigen binding fragment thereof comprising a heavy chain immunoglobulin variable domain (VH) and a light chain immunoglobulin variable domain (VL), wherein: (a) the VH domain comprises an amino acid sequence of SEQ ID NO: 3; and (b) the VL domain comprises an amino acid sequence of SEQ ID NO: 1. [0080] In one aspect, the present disclosure provides a TROP-2 antibody or antigen binding fragment thereof comprising a heavy chain immunoglobulin variable domain (VH) and a light chain immunoglobulin variable domain (VL), wherein the VH comprises a VH- CDR1 sequence of SEQ ID NO: 15, a VH-CDR2 sequence of SEQ ID NO: 16, and a VH- CDR3 sequence of SEQ ID NO: 17 and the VL comprises a VL-CDR1 sequence of SEQ ID -26- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 NO: 18, a VL-CDR2 sequence of SEQ ID NO: 19, and a VL-CDR3 sequence of SEQ ID NO: 20. In one aspect, the present disclosure provides a TROP-2 antibody or antigen binding fragment thereof comprising a heavy chain immunoglobulin variable domain (VH) and a light chain immunoglobulin variable domain (VL), wherein: (a) the VH domain comprises an amino acid sequence of SEQ ID NO: 13; and (b) the VL domain comprises an amino acid sequence of SEQ ID NO: 11. [0081] In any of the above embodiments, the antibody further comprises a Fc domain of any isotype, e.g., but are not limited to, IgG (including IgG1, IgG2, IgG3, and IgG4), IgA (including IgA1 and IgA2), IgD, IgE, or IgM, and IgY. Non-limiting examples of constant region sequences include: [0082] Human IgD constant region, Uniprot: P01880 (SEQ ID NO: 21) APTKAPDVFPIISGCRHPKDNSPVVLACLITGYHPTSVTVTWYMGTQSQPQRTFPEIQRRDS YYMTSSQLSTPLQQWRQGEYKCVVQHTASKSKKEIFRWPESPKAQASSVPTAQPQAEGSL AKATTAPATTRNTGRGGEEKKKEKEKEEQEERETKTPECPSHTQPLGVYLLTPAVQDLWLR DKATFTCFVVGSDLKDAHLTWEVAGKVPTGGVEEGLLERHSNGSQSQHSRLTLPRSLWNA GTSVTCTLNHPSLPPQRLMALREPAAQAPVKLSLNLLASSDPPEAASWLLCEVSGFSPPNILL MWLEDQREVNTSGFAPARPPPQPGSTTFWAWSVLRVPAPPSPQPATYTCVVSHEDSRTLLN ASRSLEVSYVTDHGPMK [0083] Human IgG1 constant region, Uniprot: P01857 (SEQ ID NO: 22) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL YSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVF LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYR VVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQ VSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGK [0084] Human IgG2 constant region, Uniprot: P01859 (SEQ ID NO: 23) ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL YSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPP KPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSV LTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLT CLVKGFYPSDISVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV MHEALHNHYTQKSLSLSPGK [0085] Human IgG3 constant region, Uniprot: P01860 (SEQ ID NO: 24) -27- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 ASTKGPSVFPLAPCSRSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL YSLSSVVTVPSSSLGTQTYTCNVNHKPSNTKVDKRVELKTPLGDTTHTCPRCPEPKSCDTPP PCPRCPEPKSCDTPPPCPRCPEPKSCDTPPPCPRCPAPELLGGPSVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVQFKWYVDGVEVHNAKTKPREEQYNSTFRVVSVLTVLHQDWLNGKE YKCKVSNKALPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVE WESSGQPENNYNTTPPMLDSDGSFFLYSKLTVDKSRWQQGNIFSCSVMHEALHNRFTQKSL SLSPGK [0086] Human IgM constant region, Uniprot: P01871 (SEQ ID NO: 25) GSASAPTLFPLVSCENSPSDTSSVAVGCLAQDFLPDSITLSWKYKNNSDISSTRGFPSVLRGG KYAATSQVLLPSKDVMQGTDEHVVCKVQHPNGNKEKNVPLPVIAELPPKVSVFVPPRDGF FGNPRKSKLICQATGFSPRQIQVSWLREGKQVGSGVTTDQVQAEAKESGPTTYKVTSTLTIK ESDWLGQSMFTCRVDHRGLTFQQNASSMCVPDQDTAIRVFAIPPSFASIFLTKSTKLTCLVT DLTTYDSVTISWTRQNGEAVKTHTNISESHPNATFSAVGEASICEDDWNSGERFTCTVTHTD LPSPLKQTISRPKGVALHRPDVYLLPPAREQLNLRESATITCLVTGFSPADVFVQWMQRGQP LSPEKYVTSAPMPEPQAPGRYFAHSILTVSEEEWNTGETYTCVAHEALPNRVTERTVDKST GKPTLYNVSLVMSDTAGTCY [0087] Human IgG4 constant region, Uniprot: P01861 (SEQ ID NO: 26) ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL YSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPSCPAPEFLGGPSVFLFPP KPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSV LTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLT CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSV MHEALHNHYTQKSLSLSLGK [0088] Human IgA1 constant region, Uniprot: P01876 (SEQ ID NO: 27) ASPTSPKVFPLSLCSTQPDGNVVIACLVQGFFPQEPLSVTWSESGQGVTARNFPPSQDASGD LYTTSSQLTLPATQCLAGKSVTCHVKHYTNPSQDVTVPCPVPSTPPTPSPSTPPTPSPSCCHP RLSLHRPALEDLLLGSEANLTCTLTGLRDASGVTFTWTPSSGKSAVQGPPERDLCGCYSVSS VLPGCAEPWNHGKTFTCTAAYPESKTPLTATLSKSGNTFRPEVHLLPPPSEELALNELVTLT CLARGFSPKDVLVRWLQGSQELPREKYLTWASRQEPSQGTTTFAVTSILRVAAEDWKKGD TFSCMVGHEALPLAFTQKTIDRLAGKPTHVNVSVVMAEVDGTCY [0089] Human IgA2 constant region, Uniprot: P01877 (SEQ ID NO: 28) ASPTSPKVFPLSLDSTPQDGNVVVACLVQGFFPQEPLSVTWSESGQNVTARNFPPSQDASGD LYTTSSQLTLPATQCPDGKSVTCHVKHYTNPSQDVTVPCPVPPPPPCCHPRLSLHRPALEDL LLGSEANLTCTLTGLRDASGATFTWTPSSGKSAVQGPPERDLCGCYSVSSVLPGCAQPWNH -28- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 GETFTCTAAHPELKTPLTANITKSGNTFRPEVHLLPPPSEELALNELVTLTCLARGFSPKDVL VRWLQGSQELPREKYLTWASRQEPSQGTTTFAVTSILRVAAEDWKKGDTFSCMVGHEALP LAFTQKTIDRMAGKPTHVNVSVVMAEVDGTCY [0090] Human Ig kappa constant region, Uniprot: P01834 (SEQ ID NO: 29) TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSK DSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC [0091] In some embodiments, the immunoglobulin-related compositions of the present technology comprise a heavy chain constant region that is at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or is 100% identical to SEQ ID NOS: 21-28. Additionally or alternatively, in some embodiments, the immunoglobulin-related compositions of the present technology comprise a light chain constant region that is at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or is 100% identical to SEQ ID NO: 29. [0092] In some embodiments, the anti-TROP-2 immunoglobulin-related compositions of the present technology bind to an epitope within the extracellular region of human TROP-2. In certain embodiments, the epitope is a conformational epitope or non-conformational epitope. In some embodiments, the TROP-2 polypeptide has the amino acid sequence of SEQ ID NO: 30. [0093] Homo sapiens human TROP-2, GenBank Accession No. CAA54801.1 (SEQ ID NO: 30) 1 margpglapp plrlpllllv laavtghtaa qdnctcptnk mtvcspdgpg grcqcralgs 61 gmavdcstlt skclllkarm sapknartlv rpsehalvdn dglydpdcdp egrfkarqcn 121 qtsvcwcvns vgvrrtdkgd lslrcdelvr thhilidlrh rptagafnhs dldaelrrlf 181 reryrlhpkf vaavhyeqpt iqielrqnts qkaagdvdig daayyferdi kgeslfqgrg 241 gldlrvrgep lqvertliyy ldeippkfsm krltagliav ivvvvvalva gmavlvitnr 301 rksgkykkve ikelgelrke psl [0094] In another aspect, the present disclosure provides an isolated immunoglobulin- related composition (e.g., an antibody or antigen binding fragment thereof) comprising a heavy chain (HC) amino acid sequence comprising a heavy chain (HC) amino acid sequence comprising SEQ ID NO: 4, SEQ ID NO: 14, or a variant thereof having one or more conservative amino acid substitutions. Additionally or alternatively, in some embodiments, the immunoglobulin-related compositions of the present technology comprise a light chain (LC) amino acid sequence comprising SEQ ID NO: 2, SEQ ID NO: 12, or a variant thereof having one or more conservative amino acid substitutions. In some embodiments, the immunoglobulin-related compositions of the present technology comprise a HC amino acid sequence and a LC amino acid sequence selected from the group -29- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 consisting of: SEQ ID NO: 4 and SEQ ID NO: 2, and SEQ ID NO: 14 and SEQ ID NO: 12, respectively. [0095] In some embodiments, the HC and LC immunoglobulin variable domain sequences are components of the same polypeptide chain. In other embodiments, the HC and LC immunoglobulin variable domain sequences are components of different polypeptide chains. In certain embodiments, the antibody is a full-length antibody. [0096] In some embodiments, the immunoglobulin-related compositions of the present technology bind specifically to at least one TROP-2 polypeptide. In some embodiments, the immunoglobulin-related compositions of the present technology bind at least one TROP-2 polypeptide with a dissociation constant (KD) of about 10−3 M, 10−4 M, 10−5 M, 10−6 M, 10−7 M, 10−8 M, 10−9 M, 10−10 M, 10−11 M, or 10−12 M. In certain embodiments, the immunoglobulin-related compositions are monoclonal antibodies, chimeric antibodies, humanized antibodies, bispecific antibodies, or multi-specific antibodies. In some embodiments, the antibodies comprise a human antibody framework region. [0097] In certain embodiments, the immunoglobulin-related composition includes one or more of the following characteristics: (a) a light chain immunoglobulin variable domain sequence that is at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the light chain immunoglobulin variable domain sequence of any one of SEQ ID NOs: 1 or 11; and/or (b) a heavy chain immunoglobulin variable domain sequence that is at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the heavy chain immunoglobulin variable domain sequence of any one of SEQ ID NOs: 3 or 13. In another aspect, one or more amino acid residues in the immunoglobulin-related compositions provided herein are substituted with another amino acid. The substitution may be a “conservative substitution” as defined herein. [0098] In another aspect, the present disclosure provides an antibody comprising (a) a LC sequence that is at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the LC sequence present in SEQ ID NO: 2, or SEQ ID NO: 12; and/or (b) a HC sequence that is at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the HC sequence present in SEQ ID NO: 4, or SEQ ID NO: 14. [0099] In certain embodiments, the immunoglobulin-related compositions contain an IgG1 constant region comprising one or more amino acid substitutions selected from the group consisting of N297A and K322A. Additionally or alternatively, in some -30- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 embodiments, the immunoglobulin-related compositions contain an IgG4 constant region comprising a S228P mutation. [00100] In some aspects, the anti-TROP-2 immunoglobulin-related compositions described herein contain structural modifications to facilitate rapid binding and cell uptake and/or slow release. In some aspects, the anti-TROP-2 immunoglobulin-related composition of the present technology (e.g., an antibody) may contain a deletion in the CH2 constant heavy chain region to facilitate rapid binding and cell uptake and/or slow release. In some aspects, a Fab fragment is used to facilitate rapid binding and cell uptake and/or slow release. In some aspects, a F(ab)'2 fragment is used to facilitate rapid binding and cell uptake and/or slow release. [00101] In one aspect, the present technology provides a nucleic acid sequence encoding any of the immunoglobulin-related compositions described herein. Also disclosed herein are recombinant nucleic acid sequences encoding any of the antibodies described herein. [00102] In another aspect, the present technology provides a host cell expressing any nucleic acid sequence encoding any of the immunoglobulin-related compositions described herein. [00103] The immunoglobulin-related compositions of the present technology (e.g., an anti-TROP-2 antibody) can be monospecific, bispecific, trispecific or of greater multi- specificity. Multi-specific antibodies can be specific for different epitopes of one or more TROP-2 polypeptides or can be specific for both the TROP-2 polypeptide(s) as well as for heterologous compositions, such as a heterologous polypeptide or solid support material. See, e.g., WO 93/17715; WO 92/08802; WO 91/00360; WO 92/05793; Tutt et al., J. Immunol.147: 60-69 (1991); U.S. Pat. Nos.5,573,920, 4,474,893, 5,601,819, 4,714,681, 4,925,648; 6,106,835; Kostelny et al., J. Immunol.148: 1547-1553 (1992). In some embodiments, the immunoglobulin-related compositions are chimeric. In certain embodiments, the immunoglobulin-related compositions are humanized. [00104] The immunoglobulin-related compositions of the present technology can further be recombinantly fused to a heterologous polypeptide at the N- or C-terminus or chemically conjugated (including covalently and non-covalently conjugations) to polypeptides or other compositions. For example, the immunoglobulin-related compositions of the present technology can be recombinantly fused or conjugated to molecules useful as labels in detection assays and effector molecules such as heterologous polypeptides, drugs, or toxins. -31- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 See, e.g., WO 92/08495; WO 91/14438; WO 89/12624; U.S. Pat. No.5,314,995; and EP 0 396387. [00105] In any of the above embodiments of the immunoglobulin-related compositions of the present technology, the antibody or antigen binding fragment may be optionally conjugated to an agent selected from the group consisting of isotopes, dyes, chromagens, contrast agents, drugs, toxins, cytokines, enzymes, enzyme inhibitors, hormones, hormone antagonists, growth factors, radionuclides, metals, liposomes, nanoparticles, RNA, DNA or any combination thereof. For a chemical bond or physical bond, a functional group on the immunoglobulin-related composition typically associates with a functional group on the agent. Alternatively, a functional group on the agent associates with a functional group on the immunoglobulin-related composition. [00106] The functional groups on the agent and immunoglobulin-related composition can associate directly. For example, a functional group (e.g., a sulfhydryl group) on an agent can associate with a functional group (e.g., sulfhydryl group) on an immunoglobulin-related composition to form a disulfide. Alternatively, the functional groups can associate through a cross-linking agent (i.e., linker). Some examples of cross-linking agents are described below. The cross-linker can be attached to either the agent or the immunoglobulin-related composition. The number of agents or immunoglobulin-related compositions in a conjugate is also limited by the number of functional groups present on the other. For example, the maximum number of agents associated with a conjugate depends on the number of functional groups present on the immunoglobulin-related composition. Alternatively, the maximum number of immunoglobulin-related compositions associated with an agent depends on the number of functional groups present on the agent. [00107] In yet another embodiment, the conjugate comprises one immunoglobulin- related composition associated to one agent. In one embodiment, a conjugate comprises at least one agent chemically bonded (e.g., conjugated) to at least one immunoglobulin-related composition. The agent can be chemically bonded to an immunoglobulin-related composition by any method known to those in the art. For example, a functional group on the agent may be directly attached to a functional group on the immunoglobulin-related composition. Some examples of suitable functional groups include, for example, amino, carboxyl, sulfhydryl, maleimide, isocyanate, isothiocyanate and hydroxyl. -32- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 [00108] The agent may also be chemically bonded to the immunoglobulin-related composition by means of cross-linking agents, such as dialdehydes, carbodiimides, dimaleimides, and the like. Cross-linking agents can, for example, be obtained from Pierce Biotechnology, Inc., Rockford, Ill. The Pierce Biotechnology, Inc. web-site can provide assistance. Additional cross-linking agents include the platinum cross-linking agents described in U.S. Pat. Nos.5,580,990; 5,985,566; and 6,133,038 of Kreatech Biotechnology, B.V., Amsterdam, The Netherlands. [00109] Alternatively, the functional group on the agent and immunoglobulin-related composition can be the same. Homobifunctional cross-linkers are typically used to cross- link identical functional groups. Examples of homobifunctional cross-linkers include EGS (i.e., ethylene glycol bis[succinimidylsuccinate]), DSS (i.e., disuccinimidyl suberate), DMA (i.e., dimethyl adipimidate.2HCl), DTSSP (i.e., 3,3'- dithiobis[sulfosuccinimidylpropionate])), DPDPB (i.e., 1,4-di-[3'-(2'-pyridyldithio)- propionamido]butane), and BMH (i.e., bis-maleimidohexane). Such homobifunctional cross-linkers are also available from Pierce Biotechnology, Inc. [00110] In other instances, it may be beneficial to cleave the agent from the immunoglobulin-related composition. The web-site of Pierce Biotechnology, Inc. described above can also provide assistance to one skilled in the art in choosing suitable cross-linkers which can be cleaved by, for example, enzymes in the cell. Thus the agent can be separated from the immunoglobulin-related composition. Examples of cleavable linkers include SMPT (i.e., 4-succinimidyloxycarbonyl-methyl-a-[2-pyridyldithio]toluene), Sulfo-LC- SPDP (i.e., sulfosuccinimidyl 6-(3-[2-pyridyldithio]-propionamido)hexanoate), LC-SPDP (i.e., succinimidyl 6-(3-[2-pyridyldithio]-propionamido)hexanoate), Sulfo-LC-SPDP (i.e., sulfosuccinimidyl 6-(3-[2-pyridyldithio]-propionamido)hexanoate), SPDP (i.e., N- succinimidyl 3-[2-pyridyldithio]-propionamidohexanoate), and AEDP (i.e., 3-[(2- aminoethyl)dithio]propionic acid HCl). [00111] In another embodiment, a conjugate comprises at least one agent physically bonded with at least one immunoglobulin-related composition. Any method known to those in the art can be employed to physically bond the agents with the immunoglobulin-related compositions. For example, the immunoglobulin-related compositions and agents can be mixed together by any method known to those in the art. The order of mixing is not important. For instance, agents can be physically mixed with immunoglobulin-related compositions by any method known to those in the art. For example, the immunoglobulin- -33- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 related compositions and agents can be placed in a container and agitated, by for example, shaking the container, to mix the immunoglobulin-related compositions and agents. [00112] The immunoglobulin-related compositions can be modified by any method known to those in the art. For instance, the immunoglobulin-related composition may be modified by means of cross-linking agents or functional groups, as described above. Methods of Preparing Anti-TROP-2 Antibodies of the Present Technology [00113] General Overview. Initially, a target polypeptide is chosen to which an antibody of the present technology can be raised. For example, an antibody may be raised against the full-length TROP-2 protein, or to a portion of the extracellular domain of the TROP-2 protein. Techniques for generating antibodies directed to such target polypeptides are well known to those skilled in the art. Examples of such techniques include, for example, but are not limited to, those involving display libraries, xeno or human mice, hybridomas, and the like. Target polypeptides within the scope of the present technology include any polypeptide derived from TROP-2 protein containing the extracellular domain which is capable of eliciting an immune response. [00114] It should be understood that recombinantly engineered antibodies and antibody fragments, e.g., antibody-related polypeptides, which are directed to TROP-2 protein and fragments thereof are suitable for use in accordance with the present disclosure. [00115] Anti-TROP-2 antibodies that can be subjected to the techniques set forth herein include monoclonal and polyclonal antibodies, and antibody fragments such as Fab, Fab′, F(ab′)2, Fd, scFv, diabodies, antibody light chains, antibody heavy chains and/or antibody fragments. Methods useful for the high yield production of antibody Fv-containing polypeptides, e.g., Fab′ and F(ab′)2 antibody fragments have been described. See U.S. Pat. No.5,648,237. [00116] Generally, an antibody is obtained from an originating species. More particularly, the nucleic acid or amino acid sequence of the variable portion of the light chain, heavy chain or both, of an originating species antibody having specificity for a target polypeptide antigen is obtained. An originating species is any species which was useful to generate the antibody of the present technology or library of antibodies, e.g., rat, mouse, rabbit, chicken, monkey, human, and the like. [00117] Phage or phagemid display technologies are useful techniques to derive the antibodies of the present technology. Techniques for generating and cloning monoclonal antibodies are well known to those skilled in the art. Expression of sequences encoding antibodies of the present technology, can be carried out in E. coli. [00118] Due to the degeneracy of nucleic acid coding sequences, other sequences which encode substantially the same amino acid sequences as those of the naturally occurring proteins may be used in the practice of the present technology These include, but are not limited to, nucleic acid -34- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 sequences including all or portions of the nucleic acid sequences encoding the above polypeptides, which are altered by the substitution of different codons that encode a functionally equivalent amino acid residue within the sequence, thus producing a silent change. It is appreciated that the nucleotide sequence of an immunoglobulin according to the present technology tolerates sequence homology variations of up to 25% as calculated by standard methods (“Current Methods in Sequence Comparison and Analysis,” Macromolecule Sequencing and Synthesis, Selected Methods and Applications, pp.127-149, 1998, Alan R. Liss, Inc.) so long as such a variant forms an operative antibody which recognizes TROP-2 proteins. For example, one or more amino acid residues within a polypeptide sequence can be substituted by another amino acid of a similar polarity which acts as a functional equivalent, resulting in a silent alteration. Substitutes for an amino acid within the sequence may be selected from other members of the class to which the amino acid belongs. For example, the nonpolar (hydrophobic) amino acids include alanine, leucine, isoleucine, valine, proline, phenylalanine, tryptophan and methionine. The polar neutral amino acids include glycine, serine, threonine, cysteine, tyrosine, asparagine, and glutamine. The positively charged (basic) amino acids include arginine, lysine and histidine. The negatively charged (acidic) amino acids include aspartic acid and glutamic acid. Also included within the scope of the present technology are proteins or fragments or derivatives thereof which are differentially modified during or after translation, e.g., by glycosylation, proteolytic cleavage, linkage to an antibody molecule or other cellular ligands, etc. Additionally, an immunoglobulin encoding nucleic acid sequence can be mutated in vitro or in vivo to create and/or destroy translation, initiation, and/or termination sequences or to create variations in coding regions and/or form new restriction endonuclease sites or destroy pre-existing ones, to facilitate further in vitro modification. Any technique for mutagenesis known in the art can be used, including but not limited to in vitro site directed mutagenesis, J. Biol. Chem.253:6551, use of Tab linkers (Pharmacia), and the like. [00119] Monoclonal Antibody. In one embodiment of the present technology, the antibody is an anti-TROP-2 monoclonal antibody. For example, in some embodiments, the anti-TROP-2 monoclonal antibody may be a human or a mouse anti-TROP-2 monoclonal antibody. For preparation of monoclonal antibodies directed towards the TROP-2 protein, or derivatives, fragments, analogs or homologs thereof, any technique that provides for the production of antibody molecules by continuous cell line culture can be utilized. Such techniques include, but are not limited to, the hybridoma technique (See, e.g., Kohler & Milstein, 1975. Nature 256: 495-497); the trioma technique; the human B-cell hybridoma technique (See, e.g., Kozbor, et al., 1983. Immunol. Today 4: 72) and the EBV hybridoma technique to produce human monoclonal antibodies (See, e.g., Cole, et al., 1985. In: MONOCLONAL ANTIBODIES AND CANCER THERAPY, Alan R. Liss, Inc., pp.77-96). Human monoclonal antibodies can be utilized in the practice of the present technology and can be produced by using human hybridomas (See, e.g., Cote, et al., 1983. Proc. -35- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 Natl. Acad. Sci. USA 80: 2026-2030) or by transforming human B-cells with Epstein Barr Virus in vitro (See, e.g., Cole, et al., 1985. In: MONOCLONAL ANTIBODIES AND CANCER THERAPY, Alan R. Liss, Inc., pp.77-96). For example, a population of nucleic acids that encode regions of antibodies can be isolated. PCR utilizing primers derived from sequences encoding conserved regions of antibodies is used to amplify sequences encoding portions of antibodies from the population and then DNAs encoding antibodies or fragments thereof, such as variable domains, are reconstructed from the amplified sequences. Such amplified sequences also can be fused to DNAs encoding other proteins ^ e.g., a bacteriophage coat, or a bacterial cell surface protein ^ for expression and display of the fusion polypeptides on phage or bacteria. Amplified sequences can then be expressed and further selected or isolated based, e.g., on the affinity of the expressed antibody or fragment thereof for an antigen or epitope present on the TROP-2 protein. Alternatively, hybridomas expressing anti-TROP-2 monoclonal antibodies can be prepared by immunizing a subject and then isolating hybridomas from the subject’s spleen using routine methods. See, e.g., Milstein et al., (Galfre and Milstein, Methods Enzymol (1981) 73: 3-46). Screening the hybridomas using standard methods will produce monoclonal antibodies of varying specificity (i.e., for different epitopes) and affinity. A selected monoclonal antibody with the desired properties, e.g., TROP-2 binding, can be used as expressed by the hybridoma, it can be bound to a molecule such as polyethylene glycol (PEG) to alter its properties, or a cDNA encoding it can be isolated, sequenced and manipulated in various ways. Synthetic dendromeric trees can be added to reactive amino acid side chains, e.g., lysine, to enhance the immunogenic properties of TROP-2 protein. Also, CPG-dinucleotide techniques can be used to enhance the immunogenic properties of the TROP-2 protein. Other manipulations include substituting or deleting particular amino acyl residues that contribute to instability of the antibody during storage or after administration to a subject, and affinity maturation techniques to improve affinity of the antibody of the TROP-2 protein. [00120] Hybridoma Technique. In some embodiments, the antibody of the present technology is an anti-TROP-2 monoclonal antibody produced by a hybridoma which includes a B cell obtained from a transgenic non-human animal, e.g., a transgenic mouse, having a genome comprising a human heavy chain transgene and a light chain transgene fused to an immortalized cell. Hybridoma techniques include those known in the art and taught in Harlow et al., Antibodies: A Laboratory Manual Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, 349 (1988); Hammerling et al., Monoclonal Antibodies And T-Cell Hybridomas, 563-681 (1981). Other methods for producing hybridomas and monoclonal antibodies are well known to those of skill in the art. [00121] Phage Display Technique. As noted above, the antibodies of the present technology can be produced through the application of recombinant DNA and phage display technology. For example, anti-TROP-2 antibodies, can be prepared using various phage display methods known in -36- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 the art. In phage display methods, functional antibody domains are displayed on the surface of a phage particle which carries polynucleotide sequences encoding them. Phages with a desired binding property are selected from a repertoire or combinatorial antibody library (e.g., human or murine) by selecting directly with an antigen, typically an antigen bound or captured to a solid surface or bead. Phages used in these methods are typically filamentous phage including fd and M13 with Fab, Fv or disulfide stabilized Fv antibody domains that are recombinantly fused to either the phage gene III or gene VIII protein. In addition, methods can be adapted for the construction of Fab expression libraries (See, e.g., Huse, et al., Science 246: 1275-1281, 1989) to allow rapid and effective identification of monoclonal Fab fragments with the desired specificity for a TROP-2 polypeptide, e.g., a polypeptide or derivatives, fragments, analogs or homologs thereof. Other examples of phage display methods that can be used to make the antibodies of the present technology include those disclosed in Huston et al., Proc. Natl. Acad. Sci U.S.A., 85: 5879-5883, 1988; Chaudhary et al., Proc. Natl. Acad. Sci U.S.A., 87: 1066-1070, 1990; Brinkman et al., J. Immunol. Methods 182: 41-50, 1995; Ames et al., J. Immunol. Methods 184: 177-186, 1995; Kettleborough et al., Eur. J. Immunol.24: 952-958, 1994; Persic et al., Gene 187: 9-18, 1997; Burton et al., Advances in Immunology 57: 191-280, 1994; PCT/GB91/01134; WO 90/02809; WO 91/10737; WO 92/01047; WO 92/18619; WO 93/11236; WO 95/15982; WO 95/20401; WO 96/06213; WO 92/01047 (Medical Research Council et al.); WO 97/08320 (Morphosys); WO 92/01047 (CAT/MRC); WO 91/17271 (Affymax); and U.S. Pat. Nos.5,698,426, 5,223,409, 5,403,484, 5,580,717, 5,427,908, 5,750,753, 5,821,047, 5,571,698, 5,427,908, 5,516,637, 5,780,225, 5,658,727 and 5,733,743. Methods useful for displaying polypeptides on the surface of bacteriophage particles by attaching the polypeptides via disulfide bonds have been described by Lohning, U.S. Pat. No.6,753,136. As described in the above references, after phage selection, the antibody coding regions from the phage can be isolated and used to generate whole antibodies, including human antibodies, or any other desired antigen binding fragment, and expressed in any desired host including mammalian cells, insect cells, plant cells, yeast, and bacteria. For example, techniques to recombinantly produce Fab, Fab′ and F(ab′)2 fragments can also be employed using methods known in the art such as those disclosed in WO 92/22324; Mullinax et al., BioTechniques 12: 864-869, 1992; and Sawai et al., AJRI 34: 26-34, 1995; and Better et al., Science 240: 1041- 1043, 1988. [00122] Generally, hybrid antibodies or hybrid antibody fragments that are cloned into a display vector can be selected against the appropriate antigen in order to identify variants that maintain good binding activity, because the antibody or antibody fragment will be present on the surface of the phage or phagemid particle. See, e.g., Barbas III et al., Phage Display, A Laboratory Manual (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 2001). However, other vector formats -37- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 could be used for this process, such as cloning the antibody fragment library into a lytic phage vector (modified T7 or Lambda Zap systems) for selection and/or screening. [00123] Expression of Recombinant Anti-TROP-2 Antibodies. As noted above, the antibodies of the present technology can be produced through the application of recombinant DNA technology. Recombinant polynucleotide constructs encoding an anti-TROP-2 antibody of the present technology typically include an expression control sequence operably-linked to the coding sequences of anti-TROP-2 antibody chains, including naturally-associated or heterologous promoter regions. As such, another aspect of the technology includes vectors containing one or more nucleic acid sequences encoding an anti-TROP-2 antibody of the present technology. For recombinant expression of one or more of the polypeptides of the present technology, the nucleic acid containing all or a portion of the nucleotide sequence encoding the anti-TROP-2 antibody is inserted into an appropriate cloning vector, or an expression vector (i.e., a vector that contains the necessary elements for the transcription and translation of the inserted polypeptide coding sequence) by recombinant DNA techniques well known in the art and as detailed below. Methods for producing diverse populations of vectors have been described by Lerner et al., U.S. Pat. Nos.6,291,160 and 6,680,192. [00124] In general, expression vectors useful in recombinant DNA techniques are often in the form of plasmids. In the present disclosure, “plasmid” and “vector” can be used interchangeably as the plasmid is the most commonly used form of vector. However, the present technology is intended to include such other forms of expression vectors that are not technically plasmids, such as viral vectors (e.g., replication defective retroviruses, adenoviruses and adeno-associated viruses), which serve equivalent functions. Such viral vectors permit infection of a subject and expression of a construct in that subject. In some embodiments, the expression control sequences are eukaryotic promoter systems in vectors capable of transforming or transfecting eukaryotic host cells. Once the vector has been incorporated into the appropriate host, the host is maintained under conditions suitable for high level expression of the nucleotide sequences encoding the anti-TROP-2 antibody, and the collection and purification of the anti-TROP-2 antibody, e.g., cross-reacting anti-TROP-2 antibodies. See generally, U.S.2002/0199213. These expression vectors are typically replicable in the host organisms either as episomes or as an integral part of the host chromosomal DNA. Commonly, expression vectors contain selection markers, e.g., ampicillin-resistance or hygromycin- resistance, to permit detection of those cells transformed with the desired DNA sequences. Vectors can also encode signal peptide, e.g., pectate lyase, useful to direct the secretion of extracellular antibody fragments. See U.S. Pat. No.5,576,195. [00125] The recombinant expression vectors of the present technology comprise a nucleic acid encoding a protein with TROP-2 binding properties in a form suitable for expression of the nucleic acid in a host cell, which means that the recombinant expression vectors include one or more -38- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 regulatory sequences, selected on the basis of the host cells to be used for expression that is operably-linked to the nucleic acid sequence to be expressed. Within a recombinant expression vector, “operably-linked” is intended to mean that the nucleotide sequence of interest is linked to the regulatory sequence(s) in a manner that allows for expression of the nucleotide sequence (e.g., in an in vitro transcription/translation system or in a host cell when the vector is introduced into the host cell). The term “regulatory sequence” is intended to include promoters, enhancers and other expression control elements (e.g., polyadenylation signals). Such regulatory sequences are described, e.g., in Goeddel, GENE EXPRESSION TECHNOLOGY: METHODS IN ENZYMOLOGY 185, Academic Press, San Diego, Calif. (1990). Regulatory sequences include those that direct constitutive expression of a nucleotide sequence in many types of host cell and those that direct expression of the nucleotide sequence only in certain host cells (e.g., tissue-specific regulatory sequences). It will be appreciated by those skilled in the art that the design of the expression vector can depend on such factors as the choice of the host cell to be transformed, the level of expression of polypeptide desired, etc. Typical regulatory sequences useful as promoters of recombinant polypeptide expression (e.g., anti-TROP-2 antibody), include, e.g., but are not limited to, promoters of 3-phosphoglycerate kinase and other glycolytic enzymes. Inducible yeast promoters include, among others, promoters from alcohol dehydrogenase, isocytochrome C, and enzymes responsible for maltose and galactose utilization. In one embodiment, a polynucleotide encoding an anti-TROP-2 antibody of the present technology is operably-linked to an ara B promoter and expressible in a host cell. See U.S. Pat.5,028,530. The expression vectors of the present technology can be introduced into host cells to thereby produce polypeptides or peptides, including fusion polypeptides, encoded by nucleic acids as described herein (e.g., anti-TROP-2 antibody, etc.). [00126] Another aspect of the present technology pertains to anti-TROP-2 antibody-expressing host cells, which contain a nucleic acid encoding one or more anti-TROP-2 antibodies. The recombinant expression vectors of the present technology can be designed for expression of an anti- TROP-2 antibody in prokaryotic or eukaryotic cells. For example, an anti-TROP-2 antibody can be expressed in bacterial cells such as Escherichia coli, insect cells (using baculovirus expression vectors), fungal cells, e.g., yeast, yeast cells or mammalian cells. Suitable host cells are discussed further in Goeddel, GENE EXPRESSION TECHNOLOGY: METHODS IN ENZYMOLOGY 185, Academic Press, San Diego, Calif. (1990). Alternatively, the recombinant expression vector can be transcribed and translated in vitro, e.g., using T7 promoter regulatory sequences and T7 polymerase. Methods useful for the preparation and screening of polypeptides having a predetermined property, e.g., anti-TROP-2 antibody, via expression of stochastically generated polynucleotide sequences has been previously described. See U.S. Pat. Nos.5,763,192; 5,723,323; 5,814,476; 5,817,483; 5,824,514; 5,976,862; 6,492,107; 6,569,641. -39- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 [00127] Expression of polypeptides in prokaryotes is most often carried out in E. coli with vectors containing constitutive or inducible promoters directing the expression of either fusion or non-fusion polypeptides. Fusion vectors add a number of amino acids to a polypeptide encoded therein, usually to the amino terminus of the recombinant polypeptide. Such fusion vectors typically serve three purposes: (i) to increase expression of recombinant polypeptide; (ii) to increase the solubility of the recombinant polypeptide; and (iii) to aid in the purification of the recombinant polypeptide by acting as a ligand in affinity purification. Often, in fusion expression vectors, a proteolytic cleavage site is introduced at the junction of the fusion moiety and the recombinant polypeptide to enable separation of the recombinant polypeptide from the fusion moiety subsequent to purification of the fusion polypeptide. Such enzymes, and their cognate recognition sequences, include Factor Xa, thrombin and enterokinase. Typical fusion expression vectors include pGEX (Pharmacia Biotech Inc; Smith and Johnson, 1988. Gene 67: 31-40), pMAL (New England Biolabs, Beverly, Mass.) and pRIT5 (Pharmacia, Piscataway, N.J.) that fuse glutathione S-transferase (GST), maltose E binding polypeptide, or polypeptide A, respectively, to the target recombinant polypeptide. [00128] Examples of suitable inducible non-fusion E. coli expression vectors include pTrc (Amrann et al., (1988) Gene 69: 301-315) and pET 11d (Studier et al., GENE EXPRESSION TECHNOLOGY: METHODS IN ENZYMOLOGY 185, Academic Press, San Diego, Calif. (1990) 60-89). Methods for targeted assembly of distinct active peptide or protein domains to yield multifunctional polypeptides via polypeptide fusion has been described by Pack et al., U.S. Pat. Nos. 6,294,353; 6,692,935. One strategy to maximize recombinant polypeptide expression, e.g., an anti- TROP-2 antibody, in E. coli is to express the polypeptide in host bacteria with an impaired capacity to proteolytically cleave the recombinant polypeptide. See, e.g., Gottesman, GENE EXPRESSION TECHNOLOGY: METHODS IN ENZYMOLOGY 185, Academic Press, San Diego, Calif. (1990) 119-128. Another strategy is to alter the nucleic acid sequence of the nucleic acid to be inserted into an expression vector so that the individual codons for each amino acid are those preferentially utilized in the expression host, e.g., E. coli (See, e.g., Wada, et al., 1992. Nucl. Acids Res.20: 2111- 2118). Such alteration of nucleic acid sequences of the present technology can be carried out by standard DNA synthesis techniques. [00129] In another embodiment, the anti-TROP-2 antibody expression vector is a yeast expression vector. Examples of vectors for expression in yeast Saccharomyces cerevisiae include pYepSec1 (Baldari, et al., 1987. EMBO J.6: 229-234), pMFa (Kurjan and Herskowitz, Cell 30: 933- 943, 1982), pJRY88 (Schultz et al., Gene 54: 113-123, 1987), pYES2 (Invitrogen Corporation, San Diego, Calif.), and picZ (Invitrogen Corp, San Diego, Calif.). Alternatively, an anti-TROP-2 antibody can be expressed in insect cells using baculovirus expression vectors. Baculovirus vectors available for expression of polypeptides, e.g., anti-TROP-2 antibody, in cultured insect cells (e.g., -40- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 SF9 cells) include the pAc series (Smith, et al., Mol. Cell. Biol.3: 2156-2165, 1983) and the pVL series (Lucklow and Summers, 1989. Virology 170: 31-39). [00130] In yet another embodiment, a nucleic acid encoding an anti-TROP-2 antibody of the present technology is expressed in mammalian cells using a mammalian expression vector. Examples of mammalian expression vectors include, e.g., but are not limited to, pCDM8 (Seed, Nature 329: 840, 1987) and pMT2PC (Kaufman, et al., EMBO J.6: 187-195, 1987). When used in mammalian cells, the expression vector's control functions are often provided by viral regulatory elements. For example, commonly used promoters are derived from polyoma, adenovirus 2, cytomegalovirus, and simian virus 40. For other suitable expression systems for both prokaryotic and eukaryotic cells that are useful for expression of the anti-TROP-2 antibody of the present technology, see, e.g., Chapters 16 and 17 of Sambrook, et al., MOLECULAR CLONING: A LABORATORY MANUAL.2nd ed., Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989. [00131] In another embodiment, the recombinant mammalian expression vector is capable of directing expression of the nucleic acid in a particular cell type (e.g., tissue-specific regulatory elements). Tissue-specific regulatory elements are known in the art. Non-limiting examples of suitable tissue-specific promoters include the albumin promoter (liver-specific; Pinkert, et al., Genes Dev.1: 268-277, 1987), lymphoid-specific promoters (Calame and Eaton, Adv. Immunol.43: 235- 275, 1988), promoters of T cell receptors (Winoto and Baltimore, EMBO J.8: 729-733, 1989) and immunoglobulins (Banerji, et al., 1983. Cell 33: 729-740; Queen and Baltimore, Cell 33: 741-748, 1983.), neuron-specific promoters (e.g., the neurofilament promoter; Byrne and Ruddle, Proc. Natl. Acad. Sci. USA 86: 5473-5477, 1989), pancreas-specific promoters (Edlund, et al., 1985. Science 230: 912-916), and mammary gland-specific promoters (e.g., milk whey promoter; U.S. Pat. No. 4,873,316 and European Application Publication No.264,166). Developmentally-regulated promoters are also encompassed, e.g., the murine hox promoters (Kessel and Gruss, Science 249: 374-379, 1990) and the α-fetoprotein promoter (Campes and Tilghman, Genes Dev.3: 537-546, 1989). [00132] Another aspect of the present methods pertains to host cells into which a recombinant expression vector of the present technology has been introduced. The terms “host cell” and “recombinant host cell” are used interchangeably herein. It is understood that such terms refer not only to the particular subject cell but also to the progeny or potential progeny of such a cell. Because certain modifications may occur in succeeding generations due to either mutation or environmental influences, such progeny may not, in fact, be identical to the parent cell, but are still included within the scope of the term as used herein. -41- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 [00133] A host cell can be any prokaryotic or eukaryotic cell. For example, an anti-TROP-2 antibody can be expressed in bacterial cells such as E. coli, insect cells, yeast or mammalian cells. Mammalian cells are a suitable host for expressing nucleotide segments encoding immunoglobulins or fragments thereof. See Winnacker, From Genes To Clones, (VCH Publishers, NY, 1987). A number of suitable host cell lines capable of secreting intact heterologous proteins have been developed in the art, and include Chinese hamster ovary (CHO) cell lines, various COS cell lines, HeLa cells, L cells and myeloma cell lines. In some embodiments, the cells are non-human. Expression vectors for these cells can include expression control sequences, such as an origin of replication, a promoter, an enhancer, and necessary processing information sites, such as ribosome binding sites, RNA splice sites, polyadenylation sites, and transcriptional terminator sequences. Queen et al., Immunol. Rev.89: 49, 1986. Illustrative expression control sequences are promoters derived from endogenous genes, cytomegalovirus, SV40, adenovirus, bovine papillomavirus, and the like. Co et al., J Immunol.148: 1149, 1992. Other suitable host cells are known to those skilled in the art. [00134] Vector DNA can be introduced into prokaryotic or eukaryotic cells via conventional transformation or transfection techniques. As used herein, the terms “transformation” and “transfection” are intended to refer to a variety of art-recognized techniques for introducing foreign nucleic acid (e.g., DNA) into a host cell, including calcium phosphate or calcium chloride co- precipitation, DEAE-dextran-mediated transfection, lipofection, electroporation, biolistics or viral- based transfection. Other methods used to transform mammalian cells include the use of polybrene, protoplast fusion, liposomes, electroporation, and microinjection (See generally, Sambrook et al., Molecular Cloning). Suitable methods for transforming or transfecting host cells can be found in Sambrook, et al. (MOLECULAR CLONING: A LABORATORY MANUAL.2nd ed., Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989), and other laboratory manuals. The vectors containing the DNA segments of interest can be transferred into the host cell by well-known methods, depending on the type of cellular host. [00135] For stable transfection of mammalian cells, it is known that, depending upon the expression vector and transfection technique used, only a small fraction of cells may integrate the foreign DNA into their genome. In order to identify and select these integrants, a gene that encodes a selectable marker (e.g., resistance to antibiotics) is generally introduced into the host cells along with the gene of interest. Various selectable markers include those that confer resistance to drugs, such as G418, hygromycin and methotrexate. Nucleic acid encoding a selectable marker can be introduced into a host cell on the same vector as that encoding the anti-TROP-2 antibody or can be introduced on a separate vector. Cells stably transfected with the introduced nucleic acid can be identified by drug selection (e.g., cells that have incorporated the selectable marker gene will survive, while the other cells die). -42- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 [00136] A host cell that includes an anti-TROP-2 antibody of the present technology, such as a prokaryotic or eukaryotic host cell in culture, can be used to produce (i.e., express) recombinant anti-TROP-2 antibody. In one embodiment, the method comprises culturing the host cell (into which a recombinant expression vector encoding the anti-TROP-2 antibody has been introduced) in a suitable medium such that the anti-TROP-2 antibody is produced. In another embodiment, the method further comprises the step of isolating the anti-TROP-2 antibody from the medium or the host cell. Once expressed, collections of the anti-TROP-2 antibody, e.g., the anti-TROP-2 antibodies or the anti-TROP-2 antibody-related polypeptides are purified from culture media and host cells. The anti-TROP-2 antibody can be purified according to standard procedures of the art, including HPLC purification, column chromatography, gel electrophoresis and the like. In one embodiment, the anti-TROP-2 antibody is produced in a host organism by the method of Boss et al., U.S. Pat. No.4,816,397. Usually, anti-TROP-2 antibody chains are expressed with signal sequences and are thus released to the culture media. However, if the anti-TROP-2 antibody chains are not naturally secreted by host cells, the anti-TROP-2 antibody chains can be released by treatment with mild detergent. Purification of recombinant polypeptides is well known in the art and includes ammonium sulfate precipitation, affinity chromatography purification technique, column chromatography, ion exchange purification technique, gel electrophoresis and the like (See generally Scopes, Protein Purification (Springer-Verlag, N.Y., 1982). [00137] Polynucleotides encoding anti-TROP-2 antibodies, e.g., the anti-TROP-2 antibody coding sequences, can be incorporated in transgenes for introduction into the genome of a transgenic animal and subsequent expression in the milk of the transgenic animal. See, e.g., U.S. Pat. Nos. 5,741,957, 5,304,489, and 5,849,992. Suitable transgenes include coding sequences for light and/or heavy chains in operable linkage with a promoter and enhancer from a mammary gland specific gene, such as casein or β-lactoglobulin. For production of transgenic animals, transgenes can be microinjected into fertilized oocytes, or can be incorporated into the genome of embryonic stem cells, and the nuclei of such cells transferred into enucleated oocytes. [00138] Single-Chain Antibodies. In one embodiment, the anti-TROP-2 antibody of the present technology is a single-chain anti-TROP-2 antibody. According to the present technology, techniques can be adapted for the production of single-chain antibodies specific to a TROP-2 protein (See, e.g., U.S. Pat. No.4,946,778). Examples of techniques which can be used to produce single- chain Fvs and antibodies of the present technology include those described in U.S. Pat. Nos. 4,946,778 and 5,258,498; Huston et al., Methods in Enzymology, 203: 46-88, 1991; Shu, L. et al., Proc. Natl. Acad. Sci. USA, 90: 7995-7999, 1993; and Skerra et al., Science 240: 1038-1040, 1988. [00139] Chimeric and Humanized Antibodies. In one embodiment, the anti-TROP-2 antibody of the present technology is a chimeric anti-TROP-2 antibody. In one embodiment, the anti-TROP-2 -43- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 antibody of the present technology is a humanized anti-TROP-2 antibody. In one embodiment of the present technology, the donor and acceptor antibodies are monoclonal antibodies from different species. For example, the acceptor antibody is a human antibody (to minimize its antigenicity in a human), in which case the resulting CDR-grafted antibody is termed a “humanized” antibody. [00140] Recombinant anti-TROP-2 antibodies, such as chimeric and humanized monoclonal antibodies, comprising both human and non-human portions, can be made using standard recombinant DNA techniques, and are within the scope of the present technology. For some uses, including in vivo use of the anti-TROP-2 antibody of the present technology in humans as well as use of these agents in in vitro detection assays, it is possible to use chimeric or humanized anti- TROP-2 antibodies. Such chimeric and humanized monoclonal antibodies can be produced by recombinant DNA techniques known in the art. Such useful methods include, e.g., but are not limited to, methods described in International Application No. PCT/US86/02269; U.S. Pat. No. 5,225,539; European Patent No.184187; European Patent No.171496; European Patent No. 173494; PCT International Publication No. WO 86/01533; U.S. Pat. Nos.4,816,567; 5,225,539; European Patent No.125023; Better, et al., 1988. Science 240: 1041-1043; Liu, et al., 1987. Proc. Natl. Acad. Sci. USA 84: 3439-3443; Liu, et al., 1987. J. Immunol.139: 3521-3526; Sun, et al., 1987. Proc. Natl. Acad. Sci. USA 84: 214-218; Nishimura, et al., 1987. Cancer Res.47: 999-1005; Wood, et al., 1985. Nature 314: 446-449; Shaw, et al., 1988. J. Natl. Cancer Inst.80: 1553-1559; Morrison (1985) Science 229: 1202-1207; Oi, et al. (1986) BioTechniques 4: 214; Jones, et al., 1986. Nature 321: 552-525; Verhoeyan, et al., 1988. Science 239: 1534; Morrison, Science 229: 1202, 1985; Oi et al., BioTechniques 4: 214, 1986; Gillies et al., J. Immunol. Methods, 125: 191- 202, 1989; U.S. Pat. No.5,807,715; and Beidler, et al., 1988. J. Immunol.141: 4053-4060. For example, antibodies can be humanized using a variety of techniques including CDR-grafting (EP 0 239400; WO 91/09967; U.S. Pat. No.5,530,101; 5,585,089; 5,859,205; 6,248,516; EP460167), veneering or resurfacing (EP 0592106; EP 0519596; Padlan E. A., Molecular Immunology, 28: 489-498, 1991; Studnicka et al., Protein Engineering 7: 805-814, 1994; Roguska et al., PNAS 91: 969-973, 1994), and chain shuffling (U.S. Pat. No.5,565,332). In one embodiment, a cDNA encoding a murine anti-TROP-2 monoclonal antibody is digested with a restriction enzyme selected specifically to remove the sequence encoding the Fc constant region, and the equivalent portion of a cDNA encoding a human Fc constant region is substituted (See Robinson et al., PCT/US86/02269; Akira et al., European Patent Application 184,187; Taniguchi, European Patent Application 171,496; Morrison et al., European Patent Application 173,494; Neuberger et al., WO 86/01533; Cabilly et al. U.S. Patent No.4,816,567; Cabilly et al., European Patent Application 125,023; Better et al. (1988) Science 240: 1041-1043; Liu et al. (1987) Proc. Natl. Acad. Sci. USA 84: 3439-3443; Liu et al. (1987) J Immunol 139: 3521-3526; Sun et al. (1987) Proc. Natl. Acad. Sci. USA 84: 214- 218; Nishimura et al. (1987) Cancer Res 47: 999-1005; Wood et al. (1985) Nature 314: 446-449; -44- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 and Shaw et al. (1988) J. Natl. Cancer Inst.80: 1553-1559; U.S. Pat. No.6,180,370; U.S. Pat. Nos. 6,300,064; 6,696,248; 6,706,484; 6,828,422. [00141] In one embodiment, the present technology provides the construction of humanized anti-TROP-2 antibodies that are unlikely to induce a human anti-mouse antibody (hereinafter referred to as “HAMA”) response, while still having an effective antibody effector function. As used herein, the terms “human” and “humanized”, in relation to antibodies, relate to any antibody which is expected to elicit a therapeutically tolerable weak immunogenic response in a human subject. In one embodiment, the present technology provides for a humanized anti-TROP-2 antibodies, heavy and light chain immunoglobulins. [00142] CDR Antibodies. In some embodiments, the anti-TROP-2 antibody of the present technology is an anti-TROP-2 CDR antibody. Generally the donor and acceptor antibodies used to generate the anti-TROP-2 CDR antibody are monoclonal antibodies from different species; typically the acceptor antibody is a human antibody (to minimize its antigenicity in a human), in which case the resulting CDR-grafted antibody is termed a “humanized” antibody. The graft may be of a single CDR (or even a portion of a single CDR) within a single VH or VL of the acceptor antibody, or can be of multiple CDRs (or portions thereof) within one or both of the VH and VL. Frequently, all three CDRs in all variable domains of the acceptor antibody will be replaced with the corresponding donor CDRs, though one needs to replace only as many as necessary to permit adequate binding of the resulting CDR-grafted antibody to TROP-2 protein. Methods for generating CDR-grafted and humanized antibodies are taught by Queen et al. U.S. Pat. No.5,585,089; U.S. Pat. No.5,693,761; U.S. Pat. No.5,693,762; and Winter U.S.5,225,539; and EP 0682040. Methods useful to prepare VH and VL polypeptides are taught by Winter et al., U.S. Pat. Nos.4,816,397; 6,291,158; 6,291,159; 6,291,161; 6,545,142; EP 0368684; EP0451216; and EP0120694. [00143] After selecting suitable framework region candidates from the same family and/or the same family member, either or both the heavy and light chain variable regions are produced by grafting the CDRs from the originating species into the hybrid framework regions. Assembly of hybrid antibodies or hybrid antibody fragments having hybrid variable chain regions with regard to either of the above aspects can be accomplished using conventional methods known to those skilled in the art. For example, DNA sequences encoding the hybrid variable domains described herein (i.e., frameworks based on the target species and CDRs from the originating species) can be produced by oligonucleotide synthesis and/or PCR. The nucleic acid encoding CDR regions can also be isolated from the originating species antibodies using suitable restriction enzymes and ligated into the target species framework by ligating with suitable ligation enzymes. Alternatively, the framework regions of the variable chains of the originating species antibody can be changed by site-directed mutagenesis. -45- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 [00144] Since the hybrids are constructed from choices among multiple candidates corresponding to each framework region, there exist many combinations of sequences which are amenable to construction in accordance with the principles described herein. Accordingly, libraries of hybrids can be assembled having members with different combinations of individual framework regions. Such libraries can be electronic database collections of sequences or physical collections of hybrids. [00145] This process typically does not alter the acceptor antibody’s FRs flanking the grafted CDRs. However, one skilled in the art can sometimes improve antigen binding affinity of the resulting anti-TROP-2 CDR-grafted antibody by replacing certain residues of a given FR to make the FR more similar to the corresponding FR of the donor antibody. Suitable locations of the substitutions include amino acid residues adjacent to the CDR, or which are capable of interacting with a CDR (See, e.g., US 5,585,089, especially columns 12-16). Or one skilled in the art can start with the donor FR and modify it to be more similar to the acceptor FR or a human consensus FR. Techniques for making these modifications are known in the art. Particularly if the resulting FR fits a human consensus FR for that position, or is at least 90% or more identical to such a consensus FR, doing so may not increase the antigenicity of the resulting modified anti-TROP-2 CDR-grafted antibody significantly compared to the same antibody with a fully human FR. [00146] Fc Modifications. In some embodiments, the anti-TROP-2 antibodies of the present technology comprise a variant Fc region, wherein said variant Fc region comprises at least one amino acid modification relative to a wild-type Fc region (or the parental Fc region), such that said molecule has an altered affinity for an Fc receptor (e.g., an Fc^R), provided that said variant Fc region does not have a substitution at positions that make a direct contact with Fc receptor based on crystallographic and structural analysis of Fc-Fc receptor interactions such as those disclosed by Sondermann et al., Nature, 406:267-273 (2000). Examples of positions within the Fc region that make a direct contact with an Fc receptor such as an Fc^R, include amino acids 234-239 (hinge region), amino acids 265-269 (B/C loop), amino acids 297-299 (C7E loop), and amino acids 327- 332 (F/G) loop. [00147] In some embodiments, an anti-TROP-2 antibody of the present technology has an altered affinity for activating and/or inhibitory receptors, having a variant Fc region with one or more amino acid modifications, wherein said one or more amino acid modification is a N297 substitution with alanine, or a K322 substitution with alanine. [00148] Glycosylation Modifications. In some embodiments, anti-TROP-2 antibodies of the present technology have an Fc region with variant glycosylation as compared to a parent Fc region. In some embodiments, variant glycosylation includes the absence of fucose; in some embodiments, variant glycosylation results from expression in GnT1-deficient CHO cells. -46- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 [00149] In some embodiments, the antibodies of the present technology, may have a modified glycosylation site relative to an appropriate reference antibody that binds to an antigen of interest (e.g., TROP-2), without altering the functionality of the antibody, e.g., binding activity to the antigen. As used herein, "glycosylation sites" include any specific amino acid sequence in an antibody to which an oligosaccharide (i.e., carbohydrates containing two or more simple sugars linked together) will specifically and covalently attach. [00150] Oligosaccharide side chains are typically linked to the backbone of an antibody via either N-or O-linkages. N-linked glycosylation refers to the attachment of an oligosaccharide moiety to the side chain of an asparagine residue. O-linked glycosylation refers to the attachment of an oligosaccharide moiety to a hydroxyamino acid, e.g., serine, threonine. For example, an Fc- glycoform (hTROP-2-IgGln) that lacks certain oligosaccharides including fucose and terminal N- acetylglucosamine may be produced in special CHO cells and exhibit enhanced ADCC effector function. [00151] In some embodiments, the carbohydrate content of an immunoglobulin-related composition disclosed herein is modified by adding or deleting a glycosylation site. Methods for modifying the carbohydrate content of antibodies are well known in the art and are included within the present technology, see, e.g., U.S. Patent No.6,218,149; EP 0359096B1; U.S. Patent Publication No. US 2002/0028486; International Patent Application Publication WO 03/035835; U.S. Patent Publication No.2003/0115614; U.S. Patent No.6,218,149; U.S. Patent No.6,472,511; all of which are incorporated herein by reference in their entirety. In some embodiments, the carbohydrate content of an antibody (or relevant portion or component thereof) is modified by deleting one or more endogenous carbohydrate moieties of the antibody. In some certain embodiments, the present technology includes deleting the glycosylation site of the Fc region of an antibody, by modifying position 297 from asparagine to alanine. [00152] Engineered glycoforms may be useful for a variety of purposes, including but not limited to enhancing or reducing effector function. Engineered glycoforms may be generated by any method known to one skilled in the art, for example by using engineered or variant expression strains, by co-expression with one or more enzymes, for example N-acetylglucosaminyltransferase III (GnTIII), by expressing a molecule comprising an Fc region in various organisms or cell lines from various organisms, or by modifying carbohydrate(s) after the molecule comprising Fc region has been expressed. Methods for generating engineered glycoforms are known in the art, and include but are not limited to those described in Umana et al., 1999, Nat. Biotechnol.17: 176-180; Davies et al., 2001, Biotechnol. Bioeng.74:288-294; Shields et al., 2002, J. Biol. Chem.277:26733- 26740; Shinkawa et al., 2003, J. Biol. Chem.278:3466-3473; U.S. Patent No.6,602,684; U.S. Patent Application Serial No.10/277,370; U.S. Patent Application Serial No.10/113,929; International -47- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 Patent Application Publications WO 00/61739A1 ; WO 01/292246A1; WO 02/311140A1; WO 02/30954A1; POTILLEGENT™ technology (Biowa, Inc. Princeton, N.J.); GLYCOMAB™ glycosylation engineering technology (GLYCART biotechnology AG, Zurich, Switzerland); each of which is incorporated herein by reference in its entirety. See, e.g., International Patent Application Publication WO 00/061739; U.S. Patent Application Publication No.2003/0115614; Okazaki et al., 2004, JMB, 336: 1239-49. [00153] Fusion Proteins. In one embodiment, the anti-TROP-2 antibody of the present technology is a fusion protein. The anti-TROP-2 antibodies of the present technology, when fused to a second protein, can be used as an antigenic tag. Examples of domains that can be fused to polypeptides include not only heterologous signal sequences, but also other heterologous functional regions. The fusion does not necessarily need to be direct, but can occur through linker sequences. Moreover, fusion proteins of the present technology can also be engineered to improve characteristics of the anti-TROP-2 antibodies. For instance, a region of additional amino acids, particularly charged amino acids, can be added to the N-terminus of the anti-TROP-2 antibody to improve stability and persistence during purification from the host cell or subsequent handling and storage. Also, peptide moieties can be added to an anti-TROP-2 antibody to facilitate purification. Such regions can be removed prior to final preparation of the anti-TROP-2 antibody. The addition of peptide moieties to facilitate handling of polypeptides are familiar and routine techniques in the art. The anti-TROP-2 antibody of the present technology can be fused to marker sequences, such as a peptide which facilitates purification of the fused polypeptide. In select embodiments, the marker amino acid sequence is a hexa-histidine peptide, such as the tag provided in a pQE vector (QIAGEN, Inc., Chatsworth, Calif), among others, many of which are commercially available. As described in Gentz et al., Proc. Natl. Acad. Sci. USA 86: 821-824, 1989, for instance, hexa-histidine provides for convenient purification of the fusion protein. Another peptide tag useful for purification, the “HA” tag, corresponds to an epitope derived from the influenza hemagglutinin protein. Wilson et al., Cell 37: 767, 1984. [00154] Thus, any of these above fusion proteins can be engineered using the polynucleotides or the polypeptides of the present technology. Also, in some embodiments, the fusion proteins described herein show an increased half-life in vivo. [00155] Fusion proteins having disulfide-linked dimeric structures (due to the IgG) can be more efficient in binding and neutralizing other molecules compared to the monomeric secreted protein or protein fragment alone. Fountoulakis et al., J. Biochem.270: 3958-3964, 1995. [00156] Similarly, EP-A-O 464533 (Canadian counterpart 2045869) discloses fusion proteins comprising various portions of constant region of immunoglobulin molecules together with another human protein or a fragment thereof. In many cases, the Fc part in a fusion protein is beneficial in -48- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 therapy and diagnosis, and thus can result in, e.g., improved pharmacokinetic properties. See EP-A 0232262. Alternatively, deleting or modifying the Fc part after the fusion protein has been expressed, detected, and purified, may be desired. For example, the Fc portion can hinder therapy and diagnosis if the fusion protein is used as an antigen for immunizations. In drug discovery, e.g., human proteins, such as hTROP-2, have been fused with Fc portions for the purpose of high- throughput screening assays to identify antagonists of hTROP-2. Bennett et al., J. Molecular Recognition 8: 52-58, 1995; Johanson et al., J. Biol. Chem., 270: 9459-9471, 1995. [00157] Labeled Anti-TROP-2 antibodies. In one embodiment, the anti-TROP-2 antibody of the present technology is coupled with a label moiety, i.e., detectable group. The particular label or detectable group conjugated to the anti-TROP-2 antibody is not a critical aspect of the technology, so long as it does not significantly interfere with the specific binding of the anti-TROP-2 antibody of the present technology to the TROP-2 protein. The detectable group can be any material having a detectable physical or chemical property. Such detectable labels have been well-developed in the field of immunoassays and imaging. In general, almost any label useful in such methods can be applied to the present technology. Thus, a label is any composition detectable by spectroscopic, photochemical, biochemical, immunochemical, electrical, optical or chemical means. Labels useful in the practice of the present technology include magnetic beads (e.g., Dynabeads™), fluorescent dyes (e.g., fluorescein isothiocyanate, Texas red, rhodamine, and the like), radiolabels (e.g., 3H, 14C, 35S, 125I, 121I, 131I, 112In, 99mTc), other imaging agents such as microbubbles (for ultrasound imaging), 18F, 11C, 15O, 89Zr (for Positron emission tomography), 99mTC, 111In (for Single photon emission tomography), enzymes (e.g., horse radish peroxidase, alkaline phosphatase and others commonly used in an ELISA), and calorimetric labels such as colloidal gold or colored glass or plastic (e.g., polystyrene, polypropylene, latex, and the like) beads. Patents that describe the use of such labels include U.S. Pat. Nos.3,817,837; 3,850,752; 3,939,350; 3,996,345; 4,277,437; 4,275,149; and 4,366,241, each incorporated herein by reference in their entirety and for all purposes. See also Handbook of Fluorescent Probes and Research Chemicals (6th Ed., Molecular Probes, Inc., Eugene OR.). [00158] The label can be coupled directly or indirectly to the desired component of an assay according to methods well known in the art. As indicated above, a wide variety of labels can be used, with the choice of label depending on factors such as required sensitivity, ease of conjugation with the compound, stability requirements, available instrumentation, and disposal provisions. [00159] Non-radioactive labels are often attached by indirect means. Generally, a ligand molecule (e.g., biotin) is covalently bound to the molecule. The ligand then binds to an anti-ligand (e.g., streptavidin) molecule which is either inherently detectable or covalently bound to a signal system, such as a detectable enzyme, a fluorescent compound, or a chemiluminescent compound. A number of ligands and anti-ligands can be used. Where a ligand has a natural anti-ligand, e.g., -49- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 biotin, thyroxine, and cortisol, it can be used in conjunction with the labeled, naturally-occurring anti-ligands. Alternatively, any haptenic or antigenic compound can be used in combination with an antibody, e.g., an anti-TROP-2 antibody. [00160] The molecules can also be conjugated directly to signal generating compounds, e.g., by conjugation with an enzyme or fluorophore. Enzymes of interest as labels will primarily be hydrolases, particularly phosphatases, esterases and glycosidases, or oxidoreductases, particularly peroxidases. Fluorescent compounds useful as labeling moieties, include, but are not limited to, e.g., fluorescein and its derivatives, rhodamine and its derivatives, dansyl, umbelliferone, and the like. Chemiluminescent compounds useful as labeling moieties, include, but are not limited to, e.g., luciferin, and 2,3-dihydrophthalazinediones, e.g., luminol. For a review of various labeling or signal-producing systems which can be used, see U.S. Pat. No.4,391,904. [00161] Means of detecting labels are well known to those of skill in the art. Thus, for example, where the label is a radioactive label, means for detection include a scintillation counter or photographic film as in autoradiography. Where the label is a fluorescent label, it can be detected by exciting the fluorochrome with the appropriate wavelength of light and detecting the resulting fluorescence. The fluorescence can be detected visually, by means of photographic film, by the use of electronic detectors such as charge coupled devices (CCDs) or photomultipliers and the like. Similarly, enzymatic labels can be detected by providing the appropriate substrates for the enzyme and detecting the resulting reaction product. Finally, simple colorimetric labels can be detected simply by observing the color associated with the label. Thus, in various dipstick assays, conjugated gold often appears pink, while various conjugated beads appear the color of the bead. [00162] Some assay formats do not require the use of labeled components. For instance, agglutination assays can be used to detect the presence of the target antibodies, e.g., the anti-TROP- 2 antibodies. In this case, antigen-coated particles are agglutinated by samples comprising the target antibodies. In this format, none of the components need be labeled and the presence of the target antibody is detected by simple visual inspection. Methods for Treating DPM Using the Anti-TROP-2 Antibodies of the Present Technology [00163] In one aspect, the immunoglobulin-related compositions (e.g., antibodies or antigen binding fragments thereof) of the present technology are useful for treating diffuse pleural mesothelioma (DPM) in a subject in need thereof comprising administering to the subject an effective amount of an antibody-drug conjugate comprising an anti-TROP-2 antibody or antigen binding fragment thereof. In some embodiments, the subject is -50- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 diagnosed with or is suffering from recurrent DPM or metastatic DPM. Additionally or alternatively, in certain embodiments, the subject has received a prior anti-cancer therapy. [00164] In one aspect, the present disclosure provides a method for treating diffuse pleural mesothelioma (DPM) in a subject in need thereof comprising administering to the subject an effective amount of an antibody-drug conjugate comprising an anti-TROP-2 antibody or antigen binding fragment thereof and an effective amount of an AKT inhibitor. In another aspect, the present disclosure provides a method for re-sensitizing DPM tumors to anti-TROP-2 antibody-drug conjugate therapy comprising administering to the subject an effective amount of an antibody-drug conjugate comprising an anti-TROP-2 antibody or antigen binding fragment thereof and an effective amount of an AKT inhibitor. Examples of AKT inhibitors include, but are not limited to, samotolisib, oridonin, capivasertib, ipatasertib, miransertib, afuresertib, uprosertib, BAY1125976, MK-2206, TAS-117, GSK690693, triciribine, and perifosine. In certain embodiments, the antibody-drug conjugate and the AKT inhibitor are administered separately, simultaneously, or sequentially. [00165] Additionally or alternatively, in some embodiments, the antibody-drug conjugate comprises one or more of alkylating agents, topoisomerase inhibitors, platinum agents, taxanes, vinca agents, anti-estrogen drugs, aromatase inhibitors, ovarian suppression agents, VEGF/VEGFR inhibitors, EGF/EGFR inhibitors, PARP inhibitors, cytostatic alkaloids, cytotoxic antibiotics, antimetabolites, endocrine/hormonal agents, and bisphosphonate therapy agents. In other embodiments, the antibody-drug conjugate comprises a cytotoxic drug selected from the group consisting of an anthracycline, a camptothecin, a tubulin inhibitor, a maytansinoid, a calicheamycin, an auristatin, a nitrogen mustard, an ethylenimine derivative, an alkyl sulfonate, a nitrosourea, a triazene, a folic acid analog, a taxane, a COX-2 inhibitor, a pyrimidine analog, a purine analog, an antibiotic, an enzyme inhibitor, an epipodophyllotoxin, a platinum coordination complex, a vinca alkaloid, a substituted urea, a methyl hydrazine derivative, an adrenocortical suppressant, a hormone antagonist, an antimetabolite, an alkylating agent, an antimitotic, an anti-angiogenic agent, a tyrosine kinase inhibitor, an mTOR inhibitor, a heat shock protein (HSP90) inhibitor, a proteosome inhibitor, an HDAC inhibitor, a topoisomerase inhibitor and a pro-apoptotic agent. [00166] In certain embodiments, the antibody-drug conjugate comprises cyclophosphamide, fluorouracil (or 5-fluorouracil or 5-FU), methotrexate, edatrexate (10- -51- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 ethyl-10-deaza-aminopterin), thiotepa, carboplatin, cisplatin, taxanes, paclitaxel, protein- bound paclitaxel, docetaxel, vinorelbine, tamoxifen, raloxifene, toremifene, fulvestrant, gemcitabine, irinotecan, ixabepilone, temozolmide, topotecan, vincristine, vinblastine, eribulin, mutamycin, capecitabine, anastrozole, exemestane, letrozole, leuprolide, abarelix, buserlin, goserelin, megestrol acetate, risedronate, pamidronate, ibandronate, alendronate, denosumab, zoledronate, trastuzumab, tykerb, anthracyclines, bevacizumab, Tesirine, afatinib, aplidin, azaribine, axitinib, AVL-101, AVL-291, bendamustine, bleomycin, bortezomib, bosutinib, bryostatin-1, busulfan, calicheamycin, camptothecin, 10-hydroxy camptothecin, carmustine, celecoxib, chlorambucil, COX-2 inhibitors, SN-38, cladribine, camptothecans, crizotinib, cytarabine, dacarbazine, dasatinib, dinaciclib, dactinomycin, daunorubicin, deruxtecan, DM1, DM3, DM4, doxorubicin, 2-pyrrolinodoxorubicine (2- PDox), a pro-drug form of 2-PDox (pro-2-PDox), cyano-morpholino doxorubicin, doxorubicin glucuronide, endostatin, epirubicin glucuronide, erlotinib, estramustine, epidophyllotoxin, erlotinib, entinostat, estrogen receptor binding agents, etoposide (VP 16), etoposide glucuronide, etoposide phosphate, fmgolimod, floxuridine (FUdR), 3',5'-O- dioleoyl-FudR (FUdR-dO), fludarabine, flutamide, fame syl -protein transferase inhibitors, flavopiridol, fostamatinib, ganetespib, GDC-0834, GS-1101, gefitinib, hydroxyurea, ibrutinib, idarubicin, idelalisib, ifosfamide, imatinib, lapatinib, lenolidamide, leucovorin, LFM-A13, lomustine, mechlorethamine, melphalan, mercaptopurine, 6-mercaptopurine, mitoxantrone, mithramycin, mitomycin, mitotane, monomethylauristatin F (MMAF), monomethylauristatin D (MMAD), monomethylauristatin E (MMAE), navelbine, neratinib, nilotinib, nitrosurea, olaparib, plicomycin, procarbazine, PCI-32765, pentostatin, PSI-341, semustine, SN-38, sorafenib, streptozocin, SU11248, sunitinib, transplatinum, thalidomide, thioguanine, teniposide, uracil mustard, vatalanib, vinca alkaloids, ZD1839 or combinations thereof. [00167] Additionally or alternatively, in some embodiments, the anti-TROP-2 antibody or antigen binding fragment comprises the variable heavy domain and variable light domain of sacituzumab, or datopotamab. In some embodiments, the antibody-drug conjugate comprises sacituzumab govitecan. [00168] The compositions of the present technology may optionally be administered as a single bolus to a subject in need thereof. Alternatively, the dosing regimen may comprise multiple administrations performed at various times after the appearance of tumors. -52- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 [00169] Administration can be carried out by any suitable route, including orally, intranasally, parenterally (intravenously, intramuscularly, intraperitoneally, or subcutaneously), rectally, intracranially, intratumorally, intrathecally, or topically. Administration includes self-administration and the administration by another. It is also to be appreciated that the various modes of treatment of medical conditions as described are intended to mean “substantial”, which includes total but also less than total treatment, and wherein some biologically or medically relevant result is achieved. [00170] In any and all embodiments of the methods disclosed herein, the methods of the present technology further comprise sequentially, simultaneously or separately administering to the patient an effective amount of a corticosteroid, or an antihistamine. [00171] In some embodiments, the antibodies of the present technology comprise pharmaceutical formulations which may be administered to subjects in need thereof in one or more doses. Dosage regimens can be adjusted to provide the desired response (e.g., a therapeutic response). [00172] Typically, an effective amount of the antibody compositions of the present technology, sufficient for achieving a therapeutic effect, range from about 0.000001 mg per kilogram body weight per day to about 10,000 mg per kilogram body weight per day. Typically, the dosage ranges are from about 0.0001 mg per kilogram body weight per day to about 100 mg per kilogram body weight per day. For administration of anti-TROP-2 antibodies, the dosage ranges from about 0.0001 to 100 mg/kg, and more usually 0.01 to 5 mg/kg every week, every two weeks or every three weeks, of the subject body weight. For example, dosages can be 1 mg/kg body weight or 10 mg/kg body weight every week, every two weeks or every three weeks or within the range of 1-10 mg/kg every week, every two weeks or every three weeks. In one embodiment, a single dosage of antibody ranges from 0.1-10,000 micrograms per kg body weight. In one embodiment, antibody concentrations in a carrier range from 0.2 to 2000 micrograms per delivered milliliter. An exemplary treatment regime entails administration once per every two weeks or once a month or once every 3 to 6 months. Anti-TROP-2 antibodies may be administered on multiple occasions. Intervals between single dosages can be hourly, daily, weekly, monthly or yearly. Intervals can also be irregular as indicated by measuring blood levels of the antibody in the subject. In some methods, dosage is adjusted to achieve a serum antibody concentration in the subject of from about 75 μg/mL to about 125 μg/mL, 100 μg/mL to about 150 μg/mL, from about 125 μg/mL to about 175 μg/mL, or from about 150 μg/mL to about 200 μg/mL. -53- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 Alternatively, anti-TROP-2 antibodies can be administered as a sustained release formulation, in which case less frequent administration is required. Dosage and frequency vary depending on the half-life of the antibody in the subject. The dosage and frequency of administration can vary depending on whether the treatment is prophylactic or therapeutic. In prophylactic applications, a relatively low dosage is administered at relatively infrequent intervals over a long period of time. In therapeutic applications, a relatively high dosage at relatively short intervals is sometimes required until progression of the disease is reduced or terminated, or until the subject shows partial or complete amelioration of symptoms of disease. Thereafter, the patient can be administered a prophylactic regime. [00173] Toxicity. Optimally, an effective amount (e.g., dose) of an anti-TROP-2 antibody described herein will provide therapeutic benefit without causing substantial toxicity to the subject. Toxicity of the anti-TROP-2 antibody described herein can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., by determining the LD50 (the dose lethal to 50% of the population) or the LD100 (the dose lethal to 100% of the population). The dose ratio between toxic and therapeutic effect is the therapeutic index. The data obtained from these cell culture assays and animal studies can be used in formulating a dosage range that is not toxic for use in human. The dosage of the anti-TROP-2 antibody described herein lies within a range of circulating concentrations that include the effective dose with little or no toxicity. The dosage can vary within this range depending upon the dosage form employed and the route of administration utilized. The exact formulation, route of administration and dosage can be chosen by the individual physician in view of the subject’s condition. See, e.g., Fingl et al., In: The Pharmacological Basis of Therapeutics, Ch.1 (1975). [00174] Formulations of Pharmaceutical Compositions. According to the methods of the present technology, the anti-TROP-2 antibody can be incorporated into pharmaceutical compositions suitable for administration. The pharmaceutical compositions generally comprise recombinant or substantially purified antibody and a pharmaceutically- acceptable carrier in a form suitable for administration to a subject. Pharmaceutically- acceptable carriers are determined in part by the particular composition being administered, as well as by the particular method used to administer the composition. Accordingly, there is a wide variety of suitable formulations of pharmaceutical compositions for administering the antibody compositions (See, e.g., Remington’s Pharmaceutical Sciences, Mack Publishing Co., Easton, PA 18th ed., 1990). The pharmaceutical compositions are generally -54- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 formulated as sterile, substantially isotonic and in full compliance with all Good Manufacturing Practice (GMP) regulations of the U.S. Food and Drug Administration. [00175] The terms “pharmaceutically-acceptable,” “physiologically-tolerable,” and grammatical variations thereof, as they refer to compositions, carriers, diluents and reagents, are used interchangeably and represent that the materials are capable of administration to or upon a subject without the production of undesirable physiological effects to a degree that would prohibit administration of the composition. For example, “pharmaceutically- acceptable excipient” means an excipient that is useful in preparing a pharmaceutical composition that is generally safe, non-toxic, and desirable, and includes excipients that are acceptable for veterinary use as well as for human pharmaceutical use. Such excipients can be solid, liquid, semisolid, or, in the case of an aerosol composition, gaseous. “Pharmaceutically-acceptable salts and esters” means salts and esters that are pharmaceutically-acceptable and have the desired pharmacological properties. Such salts include salts that can be formed where acidic protons present in the composition are capable of reacting with inorganic or organic bases. Suitable inorganic salts include those formed with the alkali metals, e.g., sodium and potassium, magnesium, calcium, and aluminum. Suitable organic salts include those formed with organic bases such as the amine bases, e.g., ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, and the like. Such salts also include acid addition salts formed with inorganic acids (e.g., hydrochloric and hydrobromic acids) and organic acids (e.g., acetic acid, citric acid, maleic acid, and the alkane- and arene-sulfonic acids such as methanesulfonic acid and benzenesulfonic acid). Pharmaceutically-acceptable esters include esters formed from carboxy, sulfonyloxy, and phosphonoxy groups present in the anti-TROP-2 antibody, e.g., C1-6 alkyl esters. When there are two acidic groups present, a pharmaceutically-acceptable salt or ester can be a mono-acid-mono-salt or ester or a di-salt or ester; and similarly where there are more than two acidic groups present, some or all of such groups can be salified or esterified. An anti-TROP-2 antibody named in this technology can be present in unsalified or unesterified form, or in salified and/or esterified form, and the naming of such anti- TROP-2 antibody is intended to include both the original (unsalified and unesterified) compound and its pharmaceutically-acceptable salts and esters. Also, certain embodiments of the present technology can be present in more than one stereoisomeric form, and the naming of such anti-TROP-2 antibody is intended to include all single stereoisomers and all mixtures (whether racemic or otherwise) of such stereoisomers. A person of ordinary skill -55- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 in the art, would have no difficulty determining the appropriate timing, sequence and dosages of administration for particular drugs and compositions of the present technology. [00176] Examples of such carriers or diluents include, but are not limited to, water, saline, Ringer's solutions, dextrose solution, and 5% human serum albumin. Liposomes and non-aqueous vehicles such as fixed oils may also be used. The use of such media and compounds for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or compound is incompatible with the anti-TROP-2 antibody, use thereof in the compositions is contemplated. Supplementary active compounds can also be incorporated into the compositions. [00177] A pharmaceutical composition of the present technology is formulated to be compatible with its intended route of administration. The anti-TROP-2 antibody compositions of the present technology can be administered by parenteral, topical, intravenous, oral, subcutaneous, intraarterial, intradermal, transdermal, rectal, intracranial, intrathecal, intraperitoneal, intranasal; or intramuscular routes, or as inhalants. The anti- TROP-2 antibody can optionally be administered in combination with other agents that are at least partly effective in treating DPM. [00178] Solutions or suspensions used for parenteral, intradermal, or subcutaneous application can include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial compounds such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating compounds such as ethylenediaminetetraacetic acid (EDTA); buffers such as acetates, citrates or phosphates, and compounds for the adjustment of tonicity such as sodium chloride or dextrose. The pH can be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide. The parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic. [00179] Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor ELTM (BASF, Parsippany, N.J.) or phosphate buffered saline (PBS). In all cases, the composition must be sterile and should be fluid to the extent that easy syringeability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and -56- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 fungi. The carrier can be a solvent or dispersion medium containing, e.g., water, ethanol, polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof. The proper fluidity can be maintained, e.g., by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal compounds, e.g., parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, it will be desirable to include isotonic compounds, e.g., sugars, polyalcohols such as manitol, sorbitol, sodium chloride in the composition. Prolonged absorption of the injectable compositions can be brought about by including in the composition a compound which delays absorption, e.g., aluminum monostearate and gelatin. [00180] Sterile injectable solutions can be prepared by incorporating an anti-TROP-2 antibody of the present technology in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the anti-TROP-2 antibody into a sterile vehicle that contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, methods of preparation are vacuum drying and freeze-drying that yields a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof. The antibodies of the present technology can be administered in the form of a depot injection or implant preparation which can be formulated in such a manner as to permit a sustained or pulsatile release of the active ingredient. [00181] Oral compositions generally include an inert diluent or an edible carrier. They can be enclosed in gelatin capsules or compressed into tablets. For the purpose of oral therapeutic administration, the anti-TROP-2 antibody can be incorporated with excipients and used in the form of tablets, troches, or capsules. Oral compositions can also be prepared using a fluid carrier for use as a mouthwash, wherein the compound in the fluid carrier is applied orally and swished and expectorated or swallowed. Pharmaceutically compatible binding compounds, and/or adjuvant materials can be included as part of the composition. The tablets, pills, capsules, troches and the like can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating compound such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium -57- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 stearate or Sterotes; a glidant such as colloidal silicon dioxide; a sweetening compound such as sucrose or saccharin; or a flavoring compound such as peppermint, methyl salicylate, or orange flavoring. [00182] For administration by inhalation, the anti-TROP-2 antibody is delivered in the form of an aerosol spray from pressured container or dispenser which contains a suitable propellant, e.g., a gas such as carbon dioxide, or a nebulizer. [00183] Systemic administration can also be by transmucosal or transdermal means. For transmucosal or transdermal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art, and include, e.g., for transmucosal administration, detergents, bile salts, and fusidic acid derivatives. Transmucosal administration can be accomplished through the use of nasal sprays or suppositories. For transdermal administration, the anti-TROP-2 antibody is formulated into ointments, salves, gels, or creams as generally known in the art. [00184] The anti-TROP-2 antibody can also be prepared as pharmaceutical compositions in the form of suppositories (e.g., with conventional suppository bases such as cocoa butter and other glycerides) or retention enemas for rectal delivery. [00185] In one embodiment, the anti-TROP-2 antibody is prepared with carriers that will protect the anti-TROP-2 antibody against rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Methods for preparation of such formulations will be apparent to those skilled in the art. The materials can also be obtained commercially from Alza Corporation and Nova Pharmaceuticals, Inc. Liposomal suspensions (including liposomes targeted to infected cells with monoclonal antibodies to viral antigens) can also be used as pharmaceutically- acceptable carriers. These can be prepared according to methods known to those skilled in the art, e.g., as described in U.S. Pat. No.4,522,811. Kits of the Present Technology [00186] The present technology provides kits for the treatment of DPM, comprising at least one immunoglobulin-related composition of the present technology (e.g., any antibody or antigen binding fragment described herein), or a functional variant (e.g., substitutional variant) thereof. Optionally, the above described components of the kits of the present technology are packed in suitable containers and labeled for treatment of DPM. The above- -58- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 mentioned components may be stored in unit or multi-dose containers, for example, sealed ampoules, vials, bottles, syringes, and test tubes, as an aqueous, preferably sterile, solution or as a lyophilized, preferably sterile, formulation for reconstitution. The kit may further comprise a second container which holds a diluent suitable for diluting the pharmaceutical composition towards a higher volume. Suitable diluents include, but are not limited to, the pharmaceutically acceptable excipient of the pharmaceutical composition and a saline solution. Furthermore, the kit may comprise instructions for diluting the pharmaceutical composition and/or instructions for administering the pharmaceutical composition, whether diluted or not. The containers may be formed from a variety of materials such as glass or plastic and may have a sterile access port (for example, the container may be an intravenous solution bag or a vial having a stopper which may be pierced by a hypodermic injection needle). The kit may further comprise more containers comprising a pharmaceutically acceptable buffer, such as phosphate-buffered saline, Ringer's solution and dextrose solution. It may further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles, syringes, culture medium for one or more of the suitable hosts. The kits may optionally include instructions customarily included in commercial packages of therapeutic or diagnostic products, that contain information about, for example, the indications, usage, dosage, manufacture, administration, contraindications and/or warnings concerning the use of such therapeutic or diagnostic products. [00187] The kit can also comprise, e.g., a buffering agent, a preservative or a protein- stabilizing agent. Each component of the kit can be enclosed within an individual container and all of the various containers can be within a single package, along with instructions for using the kit. The kits of the present technology may contain a written product on or in the kit container. The written product describes how to use the reagents contained in the kit, e.g., for treatment of DPM in a subject in need thereof. In certain embodiments, the use of the reagents can be according to the methods of the present technology. EXAMPLES [00188] The present technology is further illustrated by the following Examples, which should not be construed as limiting in any way. -59- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 Example 1: Materials and Methods [00189] Study design: The main purpose of this study was to assess the pro-oncogenic role of TROP-2 (TACTSD2) in diffuse pleural mesothelioma (DPM), to study the efficacy of exploiting this membrane protein as a therapeutic target in this setting and to provide preliminary data on its theranostic potential. TROP-2 expression was analyzed in independent DPM clinical cohorts at the mRNA and protein levels and correlated to different clinical features including patient survival. Isogenic cell lines were generated to perform a functional study of the effect of the modulation of TROP-2 expression on tumorigenic characteristics in vitro and in vivo. An array of patient-derived xenograft models were treated with a TROP-2-targeting antibody-drug conjugate in vivo to assess its efficacy at controlling tumor growth, and compare it to that of standard of care cytotoxin therapy. Endpoints or each treatment group were defined as the time when group average size would reach 1000mm3 of volume. Where specified, tumors were collected, and molecular analyses were performed on then to identify and validate mechanisms of resistance to treatment. All experiments were randomized and blinded where possible. Sample sizes were determined on the basis of expected effect sizes from pilot experiments and previous experience with the models used. Overall, group sizes of five or more mice were leveraged. Differences in tumor volume were tested using Student’s t-test (two- tailed), correcting for multiple measurements. All in vitro experiments were performed at least in biological triplicates each including technical triplicates. [00190] Cell lines: H2452 (CRL-5946, RRID:CVCL_1553), H28 (CRL-5820, RRID:CVCL_1555), and MSTO-211H (CRL-2081, RRID:CVCL_1430) were purchased from ATCC. Cell lines were authenticated through the Short Tandem Repeat (STR) characterization method and regularly tested for mycoplasma (Universal Mycoplasma Detection Kit, #30-1012K, ATCC). All experiments were performed in low passage cells. All cell lines were cultured according to ATCC guidelines. [00191] Plasmid vectors, virus production and transductions: Lentiviral particles were produced and used to infect isogenic cell lines of interest, as described previously (50). Lentiviral vectors used include Cas9 (#125592, Addgene, RRID:Addgene_125592), LV04 vectors expressing sgRNAs for CDC7 (#HSPD0000024539, Sigma) or the respective control vector expressing a safe targeting sgRNA BFP (#HSCONTROL_AAVS1 on LV04, Sigma), or the Lv105 vector overexpressing TACSTD2 (#EX- G0457-Lv105-B Genecopoeia). No clone isolation/expansion was doing after transduction with either Cas9, -60- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 overexpression of sgRNA plasmids, thus generating polyclonal cell cultures for each of the cell lines generated. [00192] Lentiviral particles were produced and used to infect isogenic cell lines of interest, as described previously (57), through concurrent transfection of HEK293T cells (ATCC, # CRL-1573, RRID:CVCL_0045) with a 3:2:1 ratio of lentiviral plasmid:psPAX2:pMD2.G with JetPrime transfection reagent (Polyplus, # 114-15) at a 2:1 JetPrime:DNA ratio. Medium was changed 24 h after transfection and viral supernatants were collected 72 h after transfection. Viral supernatants were syringe-filtered with a 0.45- μM PVDF filter (Millipore, # SLHVM33RS) and concentrated approximately 20-fold with Lenti-X Concentrator (Takara Bio, # 631232) according to the manufacturer’s protocol. [00193] To generate Cas9-expressing cell lines, cells were spin-transduced with lentiviral particles made out of a lentiviral plasmid designed to constitutively express Cas9 (#125592, Addgene, RRID:Addgene_125592) as described in (58), and selected with blasticidin 2.5 μg/mL. Cells were similarly spin-transduced as described in (1) with lentiviral particles made out of lentiviral LV04 vectors expressing sgRNAs for CDC7 (#HSPD0000024539, Sigma) or the respective control vector expressing a safe targeting sgRNA BFP (#HSCONTROL_AAVS1 on LV04, Sigma), or the Lv105 vector overexpressing TACSTD2 (#EX-G0457-Lv105-B Genecopoeia). [00194] Synergy assays: Cells were seeded in 96-well plates (1000 cells/well) and treated with an interval of concentrations of SG and samotolisib (Selleck Chemicals, #S8322) for 5 days. Then, cell viability was assessed with CellTiter-Glo 2.0 Assay (Promega, G9242) and normalized to the untreated wells. Synergy was calculated using the HSA method using the SynergyFinder web application (2.0, RRID:SCR_019318) (51). [00195] Immunoblotting: Protein extraction and western blot were performed from cell lines and PDXs as previously described (52). IHC was performed on formalin-fixed paraffin-embedded tissue from tumor samples, PDX slides, or TMAs derived from patients with DPM. [00196] Protein extraction and western blot were performed as previously described in (59) from frozen cell pellets or flash-frozen tumor samples using RIPA lysis buffer with 1× HALT protease inhibitor cocktail (Thermo, # 78446). Cell pellets were resuspended in five volumes of cold lysis buffer and incubated on ice for 30 min. Lysates were clarified by centrifugation at 20,000g for 10 min at 4 °C. Antibodies for TROP-2 (Cell Signaling -61- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 Technology Cat# 90540, RRID:AB_2800160), ERK (Cell Signaling Technology Cat# 4695, RRID:AB_390779), phosphor-ERK (Cell Signaling Technology Cat# 4370, RRID:AB_2315112), AKT (Cell Signaling Technology Cat# 9272, RRID:AB_329827), phospho-AKT (Cell Signaling Technology Cat# 4060, RRID:AB_2315049), KLF4 (Cell Signaling Technology Cat# 12173, RRID:AB_2797840), SOX2 (Cell Signaling Technology Cat# 3579, RRID:AB_2195767), N-cadherin (Cell Signaling Technology Cat# 13116, RRID:AB_2687616), Vimentin (Cell Signaling Technology Cat# 5741, RRID:AB_10695459), CD56 (Cell Signaling Technology Cat# 99746, RRID:AB_2868490), PRAS40 (Cell Signaling Technology Cat# 2691, RRID:AB_2225033), phospho-PRAS40 (Cell Signaling Technology Cat# 13175, RRID:AB_2798140), vinculin (Cell Signaling Technology Cat# 13901, RRID:AB_2728768), tubulin (Cell Signaling Technology Cat# 3873, RRID:AB_1904178), and actin (Cell Signaling Technology Cat# 3700, RRID:AB_2242334). Quantifications were performed with the Image Studio software (Version 3.1, Li-Cor, RRID:SCR_015795). [00197] IHC was performed on FFPE tissue from tumor samples or PDX slides or tissue microarrays derived from patients with DPM, using a TROP-2 antibody (#NBP2-49166, Novus Biologicals). For immunohistochemical staining, slides were deparaffinized and steamed for 45 min in Target Retrieval Solution (Dako). Immunocomplexes were detected using PV Poly-HRP anti-mouse IgG (Leica Microsystems, #PV6114) followed by a TSA biotin amplification step (Perkin Elmer) with DAB as the chromogen. Tissue sections were counterstained with hematoxylin, and slides were digitized on a Ventana DP 200 Slide Scanner (Roche). Expression was scored in a blinded manner by pathologists, whereby the optical density level (“0” for no brown color, “1” for faint and fine brown chromogen deposition, “2” for intermediate chromogen deposition, and “3” for prominent chromogen deposition) was multiplied by the percentage of cells at each staining level, resulting in a total H-score range of 0–300. [00198] In vivo experiments: All mice were kept in specific pathogen-free animal facilities at Memorial Sloan Kettering Cancer Center (MSK), and procedures were performed in accord with the guidelines of MSK Institutional Animal Care and Use Committee under an approved protocol. Six to ten female 6-week-old NOD.Cg- Prkdc<scid> Il2rg<tm1Wjl>/SzJ (NSG) mice were subcutaneously engrafted per treatment arm and until tumors reached 100-150 mm3. At that point, mice were randomized into groups and treated with either vehicle, SG (0.5 mg/mouse i.v. BIW), gemcitabine (40mg/kg -62- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 i.p. QW) (53-55) or irinotecan (50 mg/kg i.p QW), or with the combination of SG (0.5 mg/mouse i.v. BIW) and samotolosib (5 mg/kg p.o. QDx5). Mice weights and tumor volumes were measured twice a week and mice were sacrificed when tumors reached ~1000 mm3. The number of mice per treatment arm were selected according to previous experience with the models and response to treatments. Blinding was not performed. All animal experiments were approved by the MSK Institutional Animal Care and Use Committee (#13-07-007). [00199] For intracardiac injections, a total of 0.5 million cells were injected in the left ventricle of 5 anesthetized 6–8-week-old athymic female mice (Envigo). Immediately after surgery, and then weekly, animals were injected with D-luciferin (#LUCK-5G, GoldBiotechnology) at 15 mg/kg retro- orbitally and photonic emission was imaged using the In Vivo Imaging System (IVIS, Perkin Elmer) with a collection time of 1 minute. Tumor bioluminescence was quantified by integrating the photonic flux (photons per second) through a region encircling each tumor as determined by the LIVING IMAGES software package per manufacturer’s instructions (Perkin Elmer, RRID:SCR_014247). [00200] In vitro tumorigenic surrogate assays: Surrogate assays were performed as previously described (56). Three biological replicates, each including three technical replicates were performed for each assay. [00201] Surrogate assays were performed as indicated in (60). For growth curves, multiple 96-well plates were seeded with 3,000 cells/well and cell density was quantified using a luminescent assay (CellTiter-Glo 2.0 assay, #G9242, Promega). Cell proliferation was determined by normalizing to the day 0 cell density measurement. For agar assays, the number of colonies was counted after a period of 2 weeks to 1 month after seeding. Three biological replicates (independent experiments) were performed for each assay. For each biological replicate, three technical replicates per condition were carried out. Migration and invasion assays were performed using Cultrex BME Cell invasion assay kit (#3455- 096-K, R&D Systems), following manufacturer’s instructions. Briefly, 50,000 cells were seeded per chamber on day 0 on 0% FBS media, with 10% FBS media in the bottom well, and results were collected on day 4 using a luminescent assay (CellTiter-Glo 2.0 assay, #G9242, Promega) emitted by cells in the bottom chamber (migration) or included in the matrigel mimicking the extracellular matrix. Each experiment was replicated a minimum of three times in independent assays, and the experimental condition was normalized to control condition, which was assigned a value of 1. Analysis of invasion/migration capacity was -63- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 performed by averaging values in the independent replicates and by performing a two-tailed Student’s t-test to assess for statistical significance. [00202] RNA seq analyses: For PDX samples the FASTQ files were first mapped to a hybrid genome that consists of both human and mouse sequences into one index. The reads were mapped and then any read that mapped to the mouse genome was culled. The remaining reads were converted back to a FASTQ file and mapped to the target genome using the rnaStar aligner that maps reads genomically and resolves reads across splice junctions (61). The 2-pass mapping method was used in which the reads are mapped twice (62). The first mapping pass used a list of known annotated junctions from Ensemble. Novel junctions found in the first pass were then added to the known junctions and a second mapping pass was done (on the second pass the RemoveNoncanoncial flag was used). After mapping the output SAM files were post-processed using the PICARD tools to: add read groups, AddOrReplaceReadGroups which in addition sorted the file and converted it to the compressed BAM format. The expression count matrix was then computed from the mapped reads using HTSeq (www-huber.embl.de/users/anders/HTSeq, RRID:SCR_005514) and one of several possible gene model databases. The raw count matrix generated by HTSeq was then processed using the R/Bioconductor package DESeq2 v.1.42.1 (www-huber.embl.de/users/anders/DESeq, RRID:SCR_015687) which was used to both normalize the full dataset and analyze differential expression between sample groups. Pathway analysis was performed using the R package ClusterProfiler v.4.10.1 (RRID:SCR_016884)(63) with MsigDB Hallmark pathways (https://www.gsea- msigdb.org/gsea/msigdb/) v7.0.1, RRID:SCR_016863 ) (64, 65). [00203] For cell line RNAseq analysis, transcript counts were quantified using Salmon v1.1.0 (RRID:SCR_016863)(66) from RNA-seq reads. Fastq raw reads were mapped to 25 mer indexed hg38 genome by Salmon with default settings. Transcripts were converted to genes based on Ensembl 92 (RRID:SCR_002344)(67) and normalized by size factor at gene level. The subsequent differential gene expression analyses were processed on Salmon output files using Sleuth v0.30.0 (RRID:SCR_016883) in gene mode (68). Differentially expressed genes were identified using the Wald test. Any genes with p value adjusted by Benjamini-Hochberg method less than 0.05, and beta greater than 0.5 were regarded as significant. Gene set enrichment analysis (GSEA) (64) was performed on the full list of differentially expressed genes on the previously mentioned comparisons. Gene ranking was based on p value scores calculated as - log10(p value)*(sign of beta). Gene set annotations -64- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 include all gene sets deposited in the Molecular Signatures Database (MSigDB v7.0.1, RRID:SCR_016863) (64,65) or custom curated gene lists. The significance level of enrichment was evaluated using permutation test and the p value was adjusted by Benjamini-Hochberg procedure. Gene sets were marked as significantly enriched when the adjusted p value ≤ 0.05. These analyses were executed using the R package ClusterProfiler v3.18.1 (RRID:SCR_016884)(69). [00204] Survival Analysis in the MSKCC cohort: Kaplan-Meier survival curves using overall survival, event status (dead or alive) and corresponding H score for each sample TMA were plotted using the `survminer` (RRID:SCR_021094) and `survival` (RRID:SCR_021137) R packages. To obtain an estimate of the optimal H score cutpoint for survival (H=16.5), the `cutpointr` R package was used with the parameters method = maximize_boot_metric and metric = spec_constrain. Cutpoints at H score 5, 10, 20, 25 were also plotted to compare survival curves for significance and sample distribution between the groups. Multivariate survival analysis was performed using the `survivalAnalysis` R package. Cofactors in the model were H Score (>=16.5 or <16.5, binary factor), Sex (M/F), Smoking Status (Yes/No), Epitheliod (Yes/No), Asbestos exposure (Yes/No), Age (≥ 65 or <65 as a binary factor). [00205] Clinical samples: All study subjects provided signed informed consent for biospecimen analyses under Institutional Review Board-approved protocols (#14-209, #14- 091, #12-245). The TMA was established from samples from consented patients. Benign pleural tissue was obtained from consented patients who had undergone pleural biopsies/resections for diagnoses other than mesothelioma and whose pleural specimens were negative for the evidence of malignancy at the time of pathologic review. [00206] Radiochemistry: SG was reconstituted in saline at a concentration of 10 mg/mL before buffer exchanging via Amicon 30kD spin filter into chelexed PBS at pH 8.7. Bioconjugation of DFO was achieved by a 6 molar excess of SCN-DFO by random lysine conjugation over 1 hour incubation at 37° C. DFO- SG was purified by PD-10 column before final concentration again in an Amicon 30 kD spin filter. Radiolabeling of DFO-SG with Zirconium-89 at a targeted specific activity of 10-15 µCi/µg was achieved using a 10- fold volume of HEPES to that of the 1 M Oxalic acid 89Zr-Oxalate to yield a pH of 7.4. mixture was heated to 37° C for one hour before ITLC in 50 mM EDTA pH 5.5 to confirm high radiochemical purity (>99%). MALDI-TOF MS/MS was performed on the immunoconjugate to determine degree of SG labeling. MSK_Lx13 tumor bearing mice -65- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 were injected with 20 µg [89Zr]Zr-DFO-SG and imaged on a Siemens Inveon PET/CT system at 24-, 48-, and 72-hours post injection, while mice bearing MSK_Lx606 tumors were imaged at 24-, 48-, and 120-hours post injection and images calibrated to the percent injected dose per gram. Post imaging, mice were euthanized, and the following organs harvested for gamma counting and weighing as an orthogonal method to imaging blood, heart, lung, liver, spleen, pancreas, small intestine, large intestine, kidneys, muscle, bone, and tumor. In the Lx606 mouse model a blocking study was performed with 500 µg IgG1 pre mouse alongside the 20 µg [89Zr]Zr-DFO-SG. [00207] Statistical analyses: Comparisons between two groups were performed using paired or unpaired two-tailed Student’s t test, as described in the figure legends. A p value below 0.05 was considered statistically significant (*p ≤ 0.05, **p ≤ 0.01, and ***p ≤ 0.001). N indicates the number of biological (independent) experimental repeats; graphs indicate mean values and SEM or SD, as specified in the figure legends. All in vitro experiments were replicated a minimum of three times (biological replicates). All Western blots have been replicated a minimum of two times (biological replicates). Please refer to previous sections for detailed statistical analyses of the bioinformatic data. [00208] RNA extraction: Frozen tissues or cell pellets were weighed and homogenized in RLT and nucleic acids were extracted using the AllPrep DNA/RNA Mini Kit (QIAGEN, #80204) according to the manufacturer’s instructions. RNA was eluted in nuclease-free water. [00209] Expression Analysis in TCGA Mesothelioma Cohort: TCGA expression matrices (TPM and STAR-Counts) and corresponding meta data for the “TCGA-MESO” project were downloaded using the “TCGAbiolinks” R package v2.30.4 (The Cancer Genome Atlas (RRID:SCR_003193))(70-72). Samples were divided into thirds based on expression of TACSTD2 (TPM quantification). Differential gene expression analysis (DEG) was performed between the highest and lowest groups using the DESeq2 R package (RRID:SCR_015687) with the STAR-Counts expression data. Genes were ranked according to a composite score (-log10(pvalue)*sign(log2FoldChange)) and gene set enrichment analysis was performed with the `clusterProfiler` R package using gene sets from MSigDB (RRID:SCR_016863), Kegg (https://www.genome.jp/kegg/, SCR_012773) and Gene Ontology (GO) (https://geneontology.org/, RRID:SCR_002811). Pathways were filtered for p<0.05 and manually curated based on pathway among all gene sets. Pathways -66- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 with similar biological etiology among the three gene sets were plotted together. Heatmaps for genes of interest in relevant biological pathways were plotted using DESeq2 (RRID:SCR_015687) variance stabilizing transformed counts using the pheatmap v.1.0.12 (RRID:SCR_016418) function in R. [00210] Expression Analysis in MESOMICS cohort: Gene count matrix and associated metadata for the MESOMICS cohort were obtained from https://github.com/IARCbioinfo/MESOMICS_data (73) and normalized using DESeq2 (RRID:SCR_015687). Samples were divided into thirds based on normalized expression of TACSTD2. Downstream analysis (DEG, GSEA, heatmaps) were performed similarly to the TCGA cohort. Example 2: TROP-2 expression in DPM clinical samples [00211] TROP-2 protein expression was evaluated in 227 tumor specimens by immunohistochemistry (IHC) on a patient-derived DPM tissue microarray (TMA, FIG.10), which was detected in 30% of samples (n=67 H score range: 1 – 226). In parallel, 20 benign pleural specimens submitted for TROP-2 IHC evaluation exhibited no detectable TROP-2 expression, suggesting that TROP-2 expression appears limited to tumor tissue in this setting (p < 0.0001; FIG.1A). No significant correlation was observed of TROP-2 expression with sex, smoking status, self-reported classical occupational asbestos exposure, or median age (≥ or < 65) (FIG.1B). As previously established (2, 6, 24-26), there was an association between histologic subtype and survival, with patients harboring epithelioid tumors showing improved outcomes. TROP-2 expression was associated with worse overall survival (FIGs.1C and FIGs.5A-5D), suggestive of a negative prognostic role for TROP-2 in DPM. [00212] Taken together, these results show that TROP-2 expression is associated with poor prognosis in DPM. Example 3: TROP-2 activates oncogenic and pro-metastatic signaling in DPM [00213] To explore the potential oncogenic effects of TROP-2 in DPM, two fully independent and publicly available transcriptomic cohorts were leveraged: The Cancer Genome Atlas (TCGA)(4) and MESOMICS (27). Differential gene expression (DGE) compared tumors with high (highest tertile) versus low (lowest tertile) TACSTD2 mRNA expression, followed by pathway enrichment analyses, to nominate pathways potentially dysregulated by TROP-2 expression (FIGs.2A-2B). In both datasets, TROP-2 high- -67- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 expressing tumors displayed consistent downregulation of genes involved in DNA repair, and upregulation of genes related to inflammation, including TNF and JAK/STAT signaling, suggesting an association between TROP-2 expression and a pro- inflammatory phenotype in DPM tumors. Upregulation was observed of genes involved in pro-oncogenic signaling pathways, such as MAPK and AKT, consistent with previous reports indicating that TROP-2 triggers both ERK and AKT signaling in different tumor settings (28-30). Lastly, high TROP-2-expressing samples showed upregulation of genes involved in stemness, epithelial-to-mesenchymal transition (EMT), and metastasis, suggesting a role for TROP-2 in the metastatic process of DPM tumors. The two datasets were notably concordant (FIGs.2A-2B). [00214] To assess whether TROP-2 acts as a driver of these pathways, isogenic models derived from DPM cell lines were generated. TACSTD2 was overexpressed in the H28 cell line which lacks endogenous TROP-2 expression, and alternately overexpressed or knocked out (KO) TACSTD2 in the TROP-2-expressing H2452 cell line (FIG.2C). Transcriptomic analyses of these isogenic models supported that TROP-2-induced upregulation of genes involved in MAPK and AKT signaling, in stemness, and in EMT/metastasis (FIG.2D). Western blots validated these results at the protein level, with increased phosphorylation of ERK and AKT associated with TROP-2 overexpression, as well as increased expression of EMT markers such as N-cadherin, CD56, and vimentin, and of stemness markers including KLF4 and SOX2 (FIG.2C). [00215] The tumorigenic potential of these isogenic cell lines, including an additional one with no endogenous TROP-2 expression, MSTO-211H, in which TACSTD2 was exogenously overexpressed, was functionally characterized. TROP-2 overexpression increased proliferation in the three cell lines, while TROP-2 KO reduced cell growth (FIG. 3A). Assessment of the colony formation potential in soft agar assays showed increased colony formation by cell lines with TROP-2 overexpression, and the opposite after TROP-2 KO (FIG.3B), further supporting a role for TROP-2 as a driver of stemness in DPM, consistent with the transcriptomic and protein-level results (FIGs.2A-C). Additionally, in vitro features associated with metastatic potential were assayed in these cell lines, demonstrating that TROP-2 overexpression enhances migration and invasion (FIGs.3C- 3D). Conversely, TROP-2 KO almost completely abrogated migration and invasion abilities (FIGs.3C-3D). -68- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 [00216] To assess the capacity of TROP-2 to modulate metastatic colonization in vivo, intracardiac injections were performed in immunodeficient mice of the H28 and H2452 isogenic models with TROP-2 overexpression or KO, respectively (FIGs.3E-3F). Initial luciferase activity right after intracardiac injection was assessed to ensure homogeneity in terms of cell injection per mouse in the different conditions tested (FIG.6). Accelerated development of metastasis was observed in mice receiving the TROP-2-overexpressing H28 cells, relative to mice that received their control counterpart (FIG.3E), which translated into decreased survival (FIG.3F). Conversely, TROP-2-KO H2452 exhibited delayed development of metastasis (FIG.3E) and extended survival (FIG.3F). [00217] Taken together, these results suggest that TROP-2 expression induces oncogenic signaling and a stem-like, EMT state, and exerts a pro-oncogenic role in DPM, with increased metastatic potential. Example 4: TROP-2-targeting ADC is efficacious in DPM PDXs [00218] The efficacy of the SG ADC was assessed in PDX models imaged with [89Zr]Zr- DFO-SG with therapeutic reference to gemcitabine, a common standard of care for recurrent metastatic DPM. To select an appropriate diversity of models for testing, TROP-2 expression was evaluated in the cohort of PDXs by assessing TACSTD2 expression by RNA-Seq, and TROP-2 levels by IHC and flow cytometry (FIGs.4A and 7A). Model were then selected based on an appropriate growth rate and treatment diversity(15). Flow cytometry showed superior sensitivity for TROP-2 protein detection (76% [19/25] of tested models were positive; FIG.7B) compared to IHC (13% [1/8] of tested models were positive). Flow cytometry seemed to be the most reliable and sensitive detection method for TROP-2 detection in DPM. The frequency of PDX IHC positivity was generally consistent with that observed in primary tumors (30%; FIG.1A). Eight PDX models were assessed using all three modalities. SG efficacy was tested in MSK_Lx707 (treatment naive, low TROP-2 expression), MSK_Lx13 (pre-treated, low expression), MSK_Lx307 (treatment naive, intermediate expression) and MSK_Lx606 (pre-treated, high TROP-2 expression) (FIG.4A) (15). [00219] Remarkably, SG showed efficacy superior to gemcitabine in all models, regardless of TROP-2 expression or pre-treatment status (FIG.4B), with treatment/control (T/C) values of 14% vs.72% (MSK_Lx707), 32% vs.45% (MSK_Lx307) and 10% vs. 50% (MSK_Lx606), for SG and gemcitabine, respectively, at control condition -69- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 experimental endpoint. Gemcitabine was highly toxic in mice xenografted with the MSK_Lx13 PDX tumors, inducing death of all animals before day 60 of treatment (FIG. 4B and 8A). In this model, SG induced a sustained response suppressing tumor growth beyond day 80 of treatment, with a T/C value of 34.0% at control arm experimental endpoint (FIG.4B). The efficacy of SG was also compared with that of irinotecan (the active metabolite of the SG payload, SN38) in the two PDX models derived from pre- treated DPM tumors (FIG.4B). Notably, irinotecan showed minimal efficacy, suggesting that ADC-driven internalization may be important for response. SG appeared to be well tolerated in all PDX models, including no significant decrease in body weight relative to the controls (FIG.8A). These results suggest that SG might be an attractive therapeutic approach for patients with recalcitrant DPM tumors. [00220] Accordingly, these results demonstrate that the methods of the present technology are useful for treating recalcitrant/aggressive DPM tumors in a subject in need thereof. Example 5: AKT inhibition re-sensitizes to TROP-2-targeting ADC after development of resistance [00221] Next, to study potential mechanisms of resistance to SG, transcriptomic analysis of control and SG-treated Lx307 and Lx707 tumors from the in vivo treatment was performed (FIG.4B). Downregulation of TACSTD2 mRNA expression was not observed in the SG-treated PDX models (FIG.8B), suggesting a mechanism of resistance other than suppression or genetic deletion of the target. Analysis of DGE in tumors from SG-treated versus control animals, and subsequent pathway enrichment analysis, revealed AKT signaling as one of the top-ranking pathways with genes upregulated in the SG-treated tumors, consistently for both PDX models (FIG.4C). To validate these results at the protein level, western blots of total protein extracts were performed from control and SG- treated tumors of both models (FIG.4D). Consistent with the transcriptomic results, increased levels of phosphorylated AKT and PRAS40, a downstream effector of the AKT pathway, were observed. To test the ability of AKT inhibition to enhance or prolong sensitivity of DPM cells to SG, in vitro synergy assays were performed leveraging the AKT inhibitor samotolisib, chosen based on prior experience with the compound and tolerability in a prior clinical trial (31), and has demonstrated preliminary efficacy in solid tumors (32, 33) in endogenously TROP-2-expressing H2452 and in TROP-2-overexpressing MSTO- 211H (FIG.4E) cells. These assays demonstrated synergy between the drugs in both lines -70- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 (HSA synergy scores 8.6 and 8.0, respectively). Thus, administration of the AKT/mTOR inhibitor samotolisib was assessed as a strategy to reverse resistance to SG. Mice bearing the SG- sensitive model MSK_Lx606 were treated until the development of resistance (FIG.4F). At day 160, upon recognition of emerging acquired resistance, mice from the SG-treatment arm were equally distributed into two groups: one to continue SG-treatment and one to receive the combination treatment. Addition of samotolisib fully re-established disease control with no signs of toxicity observed in the combo-treated mice as compared to their SG monotherapy-treated counterparts, as per body weight measurements (FIGs.4F and 8C). [00222] Accordingly, these results demonstrate that the methods of the present technology are useful for treating SG resistant DPM in a subject in need thereof. Example 6: Imaging TROP-2-expressing DPM models with a novel diagnostic tracer utilizing Zirconium-89; [89Zr]Zr-DFO-SG. [00223] Immuno-positron emission tomography (PET) imaging was explored as a potential approach to assessing TROP-2 expression in DPM. The utility of a tissue biopsy to assess eligibility for a therapeutic target such as TROP-2 can be limited by intra- and inter-tumoral heterogeneity. Molecular imaging using the TROP-2 antibody can directly assess pharmacodynamic parameters, including temporal biodistribution and tumor specificity of uptake. This approach could have immediate clinical utility in identifying patients most likely to benefit from TROP-2 targeted therapies such as SG. As an exploratory pilot, SG was conjugated with the PET radionuclide Zirconium-89 to produce a TROP-2 targeted immunoPET radiotracer. The radiolabeling properties of [89Zr]Zr-DFO- SG, including bioconjugation, radiolabeling, and serum stability assays were deemed satisfactory (FIGs.9A-9C). [00224] Serial immunoPET/CT imaging of [89Zr]Zr-DFO-SG in TROP-2 low- and high- expressing models confirmed the potential of [89Zr]Zr-DFO-SG as diagnostic tracer. In the MSK_Lx13 PDX tumor model (low expression), imaging showed rapid tumoral uptake; >20% injected activity per cc (IA/CC) at 24 hours post injection. In a separate model, MSK_Lx606 (high TROP-2 expression) tumor accumulation was demonstrated through 144 hours post injection. Due to the NSG background, and therefore a lack of the Fc complement (34), a blocking dose of isotype IgG1 (500 µg) was administered to approximate imaging in an immunocompetent model with ex vivo terminal biodistribution -71- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 to confirm positive tumor imaging. These data support the feasibility of [89Zr]Zr- DFO-SG development as a diagnostic tracer for quantitative detection of TROP-2-expressing DPM. (FIGs.4G-H) EQUIVALENTS [00225] The present technology is not to be limited in terms of the particular embodiments described in this application, which are intended as single illustrations of individual aspects of the present technology. Many modifications and variations of this present technology can be made without departing from its spirit and scope, as will be apparent to those skilled in the art. Functionally equivalent methods and apparatuses within the scope of the present technology, in addition to those enumerated herein, will be apparent to those skilled in the art from the foregoing descriptions. Such modifications and variations are intended to fall within the scope of the present technology. It is to be understood that this present technology is not limited to particular methods, reagents, compounds compositions or biological systems, which can, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting. [00226] In addition, where features or aspects of the disclosure are described in terms of Markush groups, those skilled in the art will recognize that the disclosure is also thereby described in terms of any individual member or subgroup of members of the Markush group. [00227] As will be understood by one skilled in the art, for any and all purposes, particularly in terms of providing a written description, all ranges disclosed herein also encompass any and all possible subranges and combinations of subranges thereof. Any listed range can be easily recognized as sufficiently describing and enabling the same range being broken down into at least equal halves, thirds, quarters, fifths, tenths, etc. As a non- limiting example, each range discussed herein can be readily broken down into a lower third, middle third and upper third, etc. As will also be understood by one skilled in the art all language such as “up to,” “at least,” “greater than,” “less than,” and the like, include the number recited and refer to ranges which can be subsequently broken down into subranges as discussed above. Finally, as will be understood by one skilled in the art, a range includes each individual member. Thus, for example, a group having 1-3 cells refers to groups -72- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 having 1, 2, or 3 cells. Similarly, a group having 1-5 cells refers to groups having 1, 2, 3, 4, or 5 cells, and so forth. [00228] All patents, patent applications, provisional applications, and publications referred to or cited herein are incorporated by reference in their entirety, including all figures and tables, to the extent they are not inconsistent with the explicit teachings of this specification. REFERENCES 1. N. J. Vogelzang, J. J. Rusthoven, J. Symanowski, C. Denham, E. Kaukel, P. Ruffie, U. Gatzemeier, M. Boyer, S. Emri, C. Manegold, C. Niyikiza, P. Paoletti, Phase III study of pemetrexed in combination with cisplatin versus cisplatin alone in patients with malignant pleural mesothelioma. J Clin Oncol 21, 2636-2644 (2003). 2. P. Baas, A. Scherpereel, A. K. Nowak, N. Fujimoto, S. Peters, A. S. Tsao, A. S. Mansfield, S. Popat, T. Jahan, S. Antonia, Y. Oulkhouir, Y. Bautista, R. Cornelissen, L. Greillier, F. Grossi, D. Kowalski, J. Rodriguez-Cid, P. Aanur, A. Oukessou, C. Baudelet, G. Zalcman, First-line nivolumab plus ipilimumab in unresectable malignant pleural mesothelioma (CheckMate 743): a multicentre, randomised, open-label, phase 3 trial. Lancet 397, 375-386 (2021). 3. A. G. Nicholson, J. L. Sauter, A. K. Nowak, H. L. Kindler, R. R. Gill, M. Remy-Jardin, S. G. Armato, 3rd, L. Fernandez-Cuesta, R. Bueno, N. Alcala, M. Foll, H. Pass, R. Attanoos, P. Baas, M. B. Beasley, L. Brcic, K. J. Butnor, L. R. Chirieac, A. Churg, P. Courtiol, S. Dacic, M. De Perrot, T. Frauenfelder, A. Gibbs, F. R. Hirsch, K. Hiroshima, A. Husain, S. Klebe, S. Lantuejoul, A. Moreira, I. Opitz, M. Perol, A. Roden, V. Roggli, A. Scherpereel, F. Tirode, H. Tazelaar, W. D. Travis, M. S. Tsao, P. van Schil, J. M. Vignaud, B. Weynand, L. Lang-Lazdunski, I. Cree, V. W. Rusch, N. Girard, F. Galateau-Salle, EURACAN/IASLC Proposals for Updating the Histologic Classification of Pleural Mesothelioma: Towards a More Multidisciplinary Approach. J Thorac Oncol 15, 29-49 (2020). 4. J. Hmeljak, F. Sanchez-Vega, K. A. Hoadley, J. Shih, C. Stewart, D. Heiman, P. Tarpey, L. Danilova, E. Drill, E. A. Gibb, R. Bowlby, R. Kanchi, H. U. Osmanbeyoglu, Y. Sekido, J. Takeshita, Y. Newton, K. Graim, M. Gupta, C. M. Gay, L. Diao, D. L. Gibbs, V. Thorsson, L. Iype, H. Kantheti, D. T. Severson, G. Ravegnini, P. Desmeules, A. A. Jungbluth, W. D. Travis, S. Dacic, L. R. Chirieac, F. Galateau-Salle, J. Fujimoto, A. N. -73- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 Husain, H. C. Silveira, V. W. Rusch, R. C. Rintoul, H. Pass, H. Kindler, M. G. Zauderer, D. J. Kwiatkowski, R. Bueno, A. S. Tsao, J. Creaney, T. Lichtenberg, K. Leraas, J. Bowen, T. R. Network, I. Felau, J. C. Zenklusen, R. Akbani, A. D. Cherniack, L. A. Byers, M. S. Noble, J. A. Fletcher, A. G. Robertson, R. Shen, H. Aburatani, B. W. Robinson, P. Campbell, M. Ladanyi, Integrative Molecular Characterization of Malignant Pleural Mesothelioma. Cancer Discov 8, 1548-1565 (2018). 5. R. Bueno, E. W. Stawiski, L. D. Goldstein, S. Durinck, A. De Rienzo, Z. Modrusan, F. Gnad, T. T. Nguyen, B. S. Jaiswal, L. R. Chirieac, D. Sciaranghella, N. Dao, C. E. Gustafson, K. J. Munir, J. A. Hackney, A. Chaudhuri, R. Gupta, J. Guillory, K. Toy, C. Ha, Y. J. Chen, J. Stinson, S. Chaudhuri, N. Zhang, T. D. Wu, D. J. Sugarbaker, F. J. de Sauvage, W. G. Richards, S. Seshagiri, Comprehensive genomic analysis of malignant pleural mesothelioma identifies recurrent mutations, gene fusions and splicing alterations. Nat Genet 48, 407-416 (2016). 6. M. G. Zauderer, A. Martin, J. Egger, H. Rizvi, M. Offin, A. Rimner, P. S. Adusumilli, V. W. Rusch, M. G. Kris, J. L. Sauter, M. Ladanyi, R. Shen, The use of a next-generation sequencing-derived machine-learning risk-prediction model (OncoCast-MPM) for malignant pleural mesothelioma: a retrospective study. Lancet Digit Health 3, e565-e576 (2021). 7. M. Offin, S. R. Yang, J. Egger, G. Jayakumaran, R. S. Spencer, J. Lopardo, G. M. Nash, A. Cercek, W. D. Travis, M. G. Kris, M. Ladanyi, J. L. Sauter, M. G. Zauderer, Molecular Characterization of Peritoneal Mesotheliomas. J Thorac Oncol 17, 455-460 (2022). 8. H. L. Kindler, S. Novello, A. Bearz, G. L. Ceresoli, J. Aerts, J. Spicer, P. Taylor, K. Nackaerts, A. Greystoke, R. Jennens, L. Calabro, J. A. Burgers, A. Santoro, S. Cedres, P. Serwatowski, S. Ponce, J. P. Van Meerbeeck, A. K. Nowak, G. Blumenschein, Jr., J. M. Siegel, L. Kasten, K. Kochert, A. O. Walter, B. H. Childs, C. Elbi, R. Hassan, D. A. Fennell, Anetumab ravtansine versus vinorelbine in patients with relapsed, mesothelin- positive malignant pleural mesothelioma (ARCS-M): a randomised, open-label phase 2 trial. Lancet Oncol 23, 540-552 (2022). 9. D. A. Fennell, A. King, S. Mohammed, A. Branson, C. Brookes, L. Darlison, A. G. Dawson, A. Gaba, M. Hutka, B. Morgan, A. Nicholson, C. Richards, P. Wells-Jordan, G. J. Murphy, A. Thomas, S. T. s. g. Mi, Rucaparib in patients with BAP1-deficient or BRCA1- -74- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 deficient mesothelioma (MiST1): an open-label, single-arm, phase 2a clinical trial. Lancet Respir Med 9, 593-600 (2021). 10. D. A. Fennell, A. King, S. Mohammed, A. Greystoke, S. Anthony, C. Poile, N. Nusrat, M. Scotland, V. Bhundia, A. Branson, C. Brookes, L. Darlison, A. G. Dawson, A. Gaba, M. Hutka, B. Morgan, A. Bajaj, C. Richards, P. Wells-Jordan, A. Thomas, S. T. s. g. Mi, Abemaciclib in patients with p16ink4A-deficient mesothelioma (MiST2): a single-arm, open-label, phase 2 trial. Lancet Oncol 23, 374-381 (2022). 11. M. G. Zauderer, P. W. Szlosarek, S. Le Moulec, S. Popat, P. Taylor, D. Planchard, A. Scherpereel, M. Koczywas, M. Forster, R. B. Cameron, T. Peikert, E. K. Argon, N. R. Michaud, A. Szanto, J. Yang, Y. Chen, V. Kansra, S. Agarwal, D. A. Fennell, EZH2 inhibitor tazemetostat in patients with relapsed or refractory, BAP1-inactivated malignant pleural mesothelioma: a multicentre, open-label, phase 2 study. Lancet Oncol 23, 758-767 (2022). 12. P. S. Adusumilli, M. G. Zauderer, I. Riviere, S. B. Solomon, V. W. Rusch, R. E. O'Cearbhaill, A. Zhu, W. Cheema, N. K. Chintala, E. Halton, J. Pineda, R. Perez-Johnston, K. S. Tan, B. Daly, J. A. Araujo Filho, D. Ngai, E. McGee, A. Vincent, C. Diamonte, J. L. Sauter, S. Modi, D. Sikder, B. Senechal, X. Wang, W. D. Travis, M. Gonen, C. M. Rudin, R. J. Brentjens, D. R. Jones, M. Sadelain, A Phase I Trial of Regional Mesothelin-Targeted CAR T-cell Therapy in Patients with Malignant Pleural Disease, in Combination with the Anti-PD-1 Agent Pembrolizumab. Cancer Discov 11, 2748-2763 (2021). 13. M. Offin, D. L. De Silva, J. L. Sauter, J. V. Egger, E. Yorke, P. S. Adusumilli, A. Rimner, V. W. Rusch, M. G. Zauderer, Multimodality Therapy in Patients With Primary Pericardial Mesothelioma. J Thorac Oncol 17, 1428-1432 (2022). 14. M. Zhang, J. L. Luo, Q. Sun, J. Harber, A. G. Dawson, A. Nakas, S. Busacca, A. J. Sharkey, D. Waller, M. T. Sheaff, C. Richards, P. Wells-Jordan, A. Gaba, C. Poile, E. Y. Baitei, A. Bzura, J. Dzialo, M. Jama, J. Le Quesne, A. Bajaj, L. Martinson, J. A. Shaw, C. Pritchard, T. Kamata, N. Kuse, L. Brannan, P. De Philip Zhang, H. Yang, G. Griffiths, G. Wilson, C. Swanton, F. Dudbridge, E. J. Hollox, D. A. Fennell, Clonal architecture in mesothelioma is prognostic and shapes the tumour microenvironment. Nat Commun 12, 1751 (2021). 15. M. Offin, J. L. Sauter, S. E. Tischfield, J. V. Egger, S. Chavan, N. S. Shah, P. Manoj, K. Ventura, V. Allaj, E. de Stanchina, W. Travis, M. Ladanyi, A. Rimner, V. W. Rusch, P. S. -75- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 Adusumilli, J. T. Poirier, M. G. Zauderer, C. M. Rudin, T. Sen, Genomic and transcriptomic analysis of a diffuse pleural mesothelioma patient-derived xenograft library. Genome Med 14, 127 (2022). 16. S. Zaman, H. Jadid, A. C. Denson, J. E. Gray, Targeting Trop-2 in solid tumors: future prospects. Onco Targets Ther 12, 1781-1790 (2019). 17. D. Fong, P. Moser, C. Krammel, J. M. Gostner, R. Margreiter, M. Mitterer, G. Gastl, G. Spizzo, High expression of TROP2 correlates with poor prognosis in pancreatic cancer. Br J Cancer 99, 1290-1295 (2008). 18. A. Jiang, X. Gao, D. Zhang, L. Zhang, H. Lu, Expression and clinical significance of the Trop-2 gene in advanced non-small cell lung carcinoma. Oncol Lett 6, 375-380 (2013). 19. S. Alberti, S. Miotti, M. Stella, C. E. Klein, M. Fornaro, S. Menard, M. I. Colnaghi, Biochemical characterization of Trop-2, a cell surface molecule expressed by human carcinomas: formal proof that the monoclonal antibodies T16 and MOv-16 recognize Trop- 2. Hybridoma 11, 539-545 (1992). 20. A. Bardia, S. A. Hurvitz, S. M. Tolaney, D. Loirat, K. Punie, M. Oliveira, A. Brufsky, S. D. Sardesai, K. Kalinsky, A. B. Zelnak, R. Weaver, T. Traina, F. Dalenc, P. Aftimos, F. Lynce, S. Diab, J. Cortes, J. O'Shaughnessy, V. Dieras, C. Ferrario, P. Schmid, L. A. Carey, L. Gianni, M. J. Piccart, S. Loibl, D. M. Goldenberg, Q. Hong, M. S. Olivo, L. M. Itri, H. S. Rugo, A. C. T. Investigators, Sacituzumab Govitecan in Metastatic Triple-Negative Breast Cancer. N Engl J Med 384, 1529-1541 (2021). 21. A. Bardia, S. M. Tolaney, K. Punie, D. Loirat, M. Oliveira, K. Kalinsky, A. Zelnak, P. Aftimos, F. Dalenc, S. Sardesai, E. Hamilton, P. Sharma, S. Recalde, E. C. Gil, T. Traina, J. O'Shaughnessy, J. Cortes, M. Tsai, L. Vahdat, V. Dieras, L. A. Carey, H. S. Rugo, D. M. Goldenberg, Q. Hong, M. Olivo, L. M. Itri, S. A. Hurvitz, Biomarker analyses in the phase III ASCENT study of sacituzumab govitecan versus chemotherapy in patients with metastatic triple-negative breast cancer. Ann Oncol 32, 1148-1156 (2021). 22. S. T. Tagawa, A. V. Balar, D. P. Petrylak, A. R. Kalebasty, Y. Loriot, A. Flechon, R. K. Jain, N. Agarwal, M. Bupathi, P. Barthelemy, P. Beuzeboc, P. Palmbos, C. E. Kyriakopoulos, D. Pouessel, C. N. Sternberg, Q. Hong, T. Goswami, L. M. Itri, P. Grivas, TROPHY-U-01: A Phase II Open-Label Study of Sacituzumab Govitecan in Patients With Metastatic Urothelial Carcinoma Progressing After Platinum-Based Chemotherapy and Checkpoint Inhibitors. J Clin Oncol 39, 2474-2485 (2021). -76- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 23. L. Hegedüs, Ö. Okumus, F. Mairinger, T. Ploenes, S. Reuter, M. Schuler, A. Welt, S. Vega-Rubin-de-Celis, D. Theegarten, A. Bankfalvi, C. Aigner, B. Hegedüs, TROP2 expression and SN38 antitumor activity in malignant pleural mesothelioma cells provide a rationale for antibody-drug conjugate therapy. Lung Cancer 178, 237-246 (2023). 24. D. Curran, T. Sahmoud, P. Therasse, J. van Meerbeeck, P. E. Postmus, G. Giaccone, Prognostic factors in patients with pleural mesothelioma: the European Organization for Research and Treatment of Cancer experience. J Clin Oncol 16, 145-152 (1998). 25. R. M. Flores, T. Routledge, V. E. Seshan, J. Dycoco, M. Zakowski, Y. Hirth, V. W. Rusch, The impact of lymph node station on survival in 348 patients with surgically resected malignant pleural mesothelioma: implications for revision of the American Joint Committee on Cancer staging system. J Thorac Cardiovasc Surg 136, 605-610 (2008). 26. R. R. Meyerhoff, C. F. Yang, P. J. Speicher, B. C. Gulack, M. G. Hartwig, T. A. D'Amico, D. H. Harpole, M. F. Berry, Impact of mesothelioma histologic subtype on outcomes in the Surveillance, Epidemiology, and End Results database. J Surg Res 196, 23- 32 (2015). 27. L. Mangiante, N. Alcala, A. Sexton-Oates, A. Di Genova, A. Gonzalez-Perez, A. Khandekar, E. N. Bergstrom, J. Kim, X. Liu, R. Blazquez-Encinas, C. Giacobi, N. Le Stang, S. Boyault, C. Cuenin, S. Tabone-Eglinger, F. Damiola, C. Voegele, M. Ardin, M. C. Michallet, L. Soudade, T. M. Delhomme, A. Poret, M. Brevet, M. C. Copin, S. Giusiano- Courcambeck, D. Damotte, C. Girard, V. Hofman, P. Hofman, J. Mouroux, C. Cohen, S. Lacomme, J. Mazieres, V. T. de Montpreville, C. Perrin, G. Planchard, N. Rousseau, I. Rouquette, C. Sagan, A. Scherpereel, F. Thivolet, J. M. Vignaud, D. Jean, A. G. S. Ilg, R. Olaso, V. Meyer, A. Boland-Auge, J. F. Deleuze, J. Altmuller, P. Nuernberg, A. Ibanez- Costa, J. P. Castano, S. Lantuejoul, A. Ghantous, C. Maussion, P. Courtiol, H. Hernandez- Vargas, C. Caux, N. Girard, N. Lopez-Bigas, L. B. Alexandrov, F. Galateau-Salle, M. Foll, L. Fernandez-Cuesta, Multiomic analysis of malignant pleural mesothelioma identifies molecular axes and specialized tumor profiles driving intertumor heterogeneity. Nat Genet 55, 607-618 (2023). 28. R. Cubas, S. Zhang, M. Li, C. Chen, Q. Yao, Trop2 expression contributes to tumor pathogenesis by activating the ERK MAPK pathway. Mol Cancer 9, 253 (2010). 29. S. Lenart, P. Lenart, J. Smarda, J. Remsik, K. Soucek, P. Benes, Trop2: Jack of All Trades, Master of None. Cancers (Basel) 12, (2020). -77- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 30. E. Guerra, M. Trerotola, R. Tripaldi, A. L. Aloisi, P. Simeone, A. Sacchetti, V. Relli, A. D'Amore, R. La Sorda, R. Lattanzio, M. Piantelli, S. Alberti, Trop-2 Induces Tumor Growth Through AKT and Determines Sensitivity to AKT Inhibitors. Clin Cancer Res 22, 4197- 4205 (2016). 31. C. J. Sweeney, I. J. Percent, S. Babu, J. L. Cultrera, B. A. Mehlhaff, O. B. Goodman, D. S. Morris, I. D. Schnadig, C. Albany, N. D. Shore, P. R. Sieber, S. C. Guba, W. Zhang, V. Wacheck, G. P. Donoho, A. M. Szpurka, S. Callies, B. K. Lin, J. C. Bendell, Phase Ib/II Study of Enzalutamide with Samotolisib (LY3023414) or Placebo in Patients with Metastatic Castration-Resistant Prostate Cancer. Clin Cancer Res 28, 2237-2247 (2022). 32. J. C. Bendell, A. M. Varghese, D. M. Hyman, T. M. Bauer, S. Pant, S. Callies, J. Lin, R. Martinez, E. Wickremsinhe, A. Fink, V. Wacheck, K. N. Moore, A First-in-Human Phase 1 Study of LY3023414, an Oral PI3K/mTOR Dual Inhibitor, in Patients with Advanced Cancer. Clin Cancer Res 24, 3253-3262 (2018). 33. M. M. Rubinstein, D. M. Hyman, I. Caird, H. Won, K. Soldan, K. Seier, A. Iasonos, W. P. Tew, R. E. O'Cearbhaill, R. N. Grisham, M. L. Hensley, T. Troso-Sandoval, P. Sabbatini, J. Guillen, S. D. Selcuklu, C. Zimel, J. Torrisi, C. Aghajanian, V. Makker, Phase 2 study of LY3023414 in patients with advanced endometrial cancer harboring activating mutations in the PI3K pathway. Cancer 126, 1274-1282 (2020). 34. S. K. Sharma, A. Chow, S. Monette, D. Vivier, J. Pourat, K. J. Edwards, T. R. Dilling, D. Abdel-Atti, B. M. Zeglis, J. T. Poirier, J. S. Lewis, Fc-Mediated Anomalous Biodistribution of Therapeutic Antibodies in Immunodeficient Mouse Models. Cancer Res 78, 1820-1832 (2018). 35. T. El Sewedy, M. Fornaro, S. Alberti, Cloning of the murine TROP2 gene: conservation of a PIP2-binding sequence in the cytoplasmic domain of TROP-2. Int J Cancer 75, 324- 330 (1998). 36. S. Miotti, S. Canevari, S. Menard, D. Mezzanzanica, G. Porro, S. M. Pupa, M. Regazzoni, E. Tagliabue, M. I. Colnaghi, Characterization of human ovarian carcinoma- associated antigens defined by novel monoclonal antibodies with tumor-restricted specificity. Int J Cancer 39, 297-303 (1987). 37. G. Muhlmann, G. Spizzo, J. Gostner, M. Zitt, H. Maier, P. Moser, G. Gastl, M. Zitt, H. M. Muller, R. Margreiter, D. Ofner, D. Fong, TROP2 expression as prognostic marker for gastric carcinoma. J Clin Pathol 62, 152-158 (2009). -78- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 38. J. C. Lin, Y. Y. Wu, J. Y. Wu, T. C. Lin, C. T. Wu, Y. L. Chang, Y. S. Jou, T. M. Hong, P. C. Yang, TROP2 is epigenetically inactivated and modulates IGF-1R signalling in lung adenocarcinoma. EMBO Mol Med 4, 472-485 (2012). 39. H. Guan, Z. Guo, W. Liang, H. Li, G. Wei, L. Xu, H. Xiao, Y. Li, Trop2 enhances invasion of thyroid cancer by inducing MMP2 through ERK and JNK pathways. BMC Cancer 17, 486 (2017). 40. P. Lombardi, M. Filetti, R. Falcone, V. Altamura, F. Paroni Sterbini, E. Bria, A. Fabi, D. Giannarelli, G. Scambia, G. Daniele, Overview of Trop-2 in Cancer: From Pre-Clinical Studies to Future Directions in Clinical Settings. Cancers (Basel) 15, (2023). 41. S. Lenart, P. Lenart, L. Knopfova, H. Kotasova, V. Pelkova, V. Sedlakova, O. Vacek, J. Pokludova, V. Can, J. Smarda, K. Soucek, A. Hampl, P. Benes, TACSTD2 upregulation is an early reaction to lung infection. Sci Rep 12, 9583 (2022). 42. Q. Liu, H. Li, Q. Wang, Y. Zhang, W. Wang, S. Dou, W. Xiao, Increased expression of TROP2 in airway basal cells potentially contributes to airway remodeling in chronic obstructive pulmonary disease. Respir Res 17, 159 (2016). 43. L. Yang, Y. Pang, H. L. Moses, TGF-beta and immune cells: an important regulatory axis in the tumor microenvironment and progression. Trends Immunol 31, 220-227 (2010). 44. A. Bessede, F. Peyraud, B. Besse, S. Cousin, M. Cabart, F. Chomy, C. Rey, O. Lara, O. Odin, I. Nafia, L. Vanhersecke, F. Barlesi, J. P. Guegan, A. Italiano, TROP2 Is Associated with Primary Resistance to Immune Checkpoint Inhibition in Patients with Advanced Non- Small Cell Lung Cancer. Clin Cancer Res 30, 779-785 (2024). 45. K. M. Tully, S. Tendler, L. M. Carter, S. K. Sharma, Z. V. Samuels, K. Mandleywala, J. A. Korsen, A. M. Delos Reyes, A. Piersigilli, W. D. Travis, T. Sen, N. Pillarsetty, J. T. Poirier, C. M. Rudin, J. S. Lewis, Radioimmunotherapy Targeting Delta-like Ligand 3 in Small Cell Lung Cancer Exhibits Antitumor Efficacy with Low Toxicity. Clin Cancer Res 28, 1391-1401 (2022). 46. J. A. Korsen, J. A. Gutierrez, K. M. Tully, L. M. Carter, Z. V. Samuels, S. Khitrov, J. T. Poirier, C. M. Rudin, Y. Chen, M. J. Morris, L. Bodei, N. Pillarsetty, J. S. Lewis, Delta-like ligand 3-targeted radioimmunotherapy for neuroendocrine prostate cancer. Proc Natl Acad Sci U S A 119, e2203820119 (2022). -79- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 47. B. T. Li, F. Michelini, S. Misale, E. Cocco, L. Baldino, Y. Cai, S. Shifman, H. Y. Tu, M. L. Myers, C. Xu, M. Mattar, I. Khodos, M. Little, B. Qeriqi, G. Weitsman, C. J. Wilhem, A. S. Lalani, I. Diala, R. A. Freedman, N. U. Lin, D. B. Solit, M. F. Berger, P. R. Barber, T. Ng, M. Offin, J. M. Isbell, D. R. Jones, H. A. Yu, S. Thyparambil, W. L. Liao, A. Bhalkikar, F. Cecchi, D. M. Hyman, J. S. Lewis, D. J. Buonocore, A. L. Ho, V. Makker, J. S. Reis-Filho, P. Razavi, M. E. Arcila, M. G. Kris, J. T. Poirier, R. Shen, J. Tsurutani, G. A. Ulaner, E. de Stanchina, N. Rosen, C. M. Rudin, M. Scaltriti, HER2-Mediated Internalization of Cytotoxic Agents in ERBB2 Amplified or Mutant Lung Cancers. Cancer Discov 10, 674-687 (2020). 48. A. Quintanal-Villalonga, H. Taniguchi, Y. Hao, A. Chow, Y. A. Zhan, S. S. Chavan, F. Uddin, V. Allaj, P. Manoj, N. S. Shah, J. M. Chan, M. Offin, M. Ciampricotti, J. Ray- Kirton, J. Egger, U. Bhanot, I. Linkov, M. Asher, M. H. Roehrl, J. Qiu, E. de Stanchina, T. J. Hollmann, R. P. Koche, T. Sen, J. T. Poirier, C. M. Rudin, Inhibition of XPO1 Sensitizes Small Cell Lung Cancer to First- and Second-Line Chemotherapy. Cancer Res 82, 472-483 (2022). 49. Y. He, M. M. Sun, G. G. Zhang, J. Yang, K. S. Chen, W. W. Xu, B. Li, Targeting PI3K/Akt signal transduction for cancer therapy. Signal Transduct Target Ther 6, 425 (2021). 50. C. H. Hulton, E. A. Costa, N. S. Shah, A. Quintanal-Villalonga, G. Heller, E. de Stanchina, C. M. Rudin, J. T. Poirier, Direct genome editing of patient-derived xenografts using CRISPR-Cas9 enables rapid in vivo functional genomics. Nat Cancer 1, 359-369 (2020). 51. A. Ianevski, A. K. Giri, T. Aittokallio, SynergyFinder 2.0: visual analytics of multi-drug combination synergies. Nucleic Acids Res 48, W488-W493 (2020). 52. A. Quintanal-Villalonga, H. Taniguchi, Y. A. Zhan, M. M. Hasan, S. S. Chavan, F. Meng, F. Uddin, P. Manoj, M. T. A. Donoghue, H. H. Won, J. M. Chan, M. Ciampricotti, A. Chow, M. Offin, J. C. Chang, J. Ray-Kirton, S. E. Tischfield, J. Egger, U. K. Bhanot, I. Linkov, M. Asher, S. Sinha, J. Silber, C. A. Iacobuzio-Donahue, M. H. Roehrl, T. J. Hollmann, H. A. Yu, J. Qiu, E. de Stanchina, M. K. Baine, N. Rekhtman, J. T. Poirier, B. Loomis, R. P. Koche, C. M. Rudin, T. Sen, Multiomic Analysis of Lung Tumors Defines Pathways Activated in Neuroendocrine Transformation. Cancer Discov 11, 3028-3047 (2021). -80- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 53. M. J. Rathos, K. Joshi, H. Khanwalkar, S. M. Manohar, K. S. Joshi, Molecular evidence for increased antitumor activity of gemcitabine in combination with a cyclin-dependent kinase inhibitor, P276-00 in pancreatic cancers. J Transl Med 10, 161 (2012). 54. W. Waissi, A. Nicol, M. Jung, M. Rousseau, D. Jarnet, G. Noel, H. Burckel, Radiosensitizing Pancreatic Cancer with PARP Inhibitor and Gemcitabine: An In Vivo and a Whole-Transcriptome Analysis after Proton or Photon Irradiation. Cancers (Basel) 13, (2021). 55. D. C. Zhang, J. L. Liu, Y. B. Ding, J. G. Xia, G. Y. Chen, Icariin potentiates the antitumor activity of gemcitabine in gallbladder cancer by suppressing NF-kappaB. Acta Pharmacol Sin 34, 301-308 (2013). 56. A. Quintanal-Villalonga, S. Molina-Pinelo, C. Cirauqui, L. Ojeda-Marquez, A. Marrugal, R. Suarez, E. Conde, S. Ponce-Aix, A. B. Enguita, A. Carnero, I. Ferrer, L. Paz- Ares, FGFR1 Cooperates with EGFR in Lung Cancer Oncogenesis, and Their Combined Inhibition Shows Improved Efficacy. J Thorac Oncol 14, 641-655 (2019). 57. C. H. Hulton, E. A. Costa, N. S. Shah, A. Quintanal-Villalonga, G. Heller, E. de Stanchina, C. M. Rudin, J. T. Poirier, Direct genome editing of patient-derived xenografts using CRISPR-Cas9 enables rapid in vivo functional genomics. Nat Cancer 1, 359-369 (2020). 58. A. Quintanal-Villalonga, V. Durani, A. Sabet, E. Redin, K. Kawasaki, M. Shafer, W. R. Karthaus, S. Zaidi, Y. A. Zhan, P. Manoj, H. Sridhar, N. S. Shah, A. Chow, U. K. Bhanot, I. Linkov, M. Asher, H. A. Yu, J. Qiu, E. de Stanchina, R. A. Patel, C. Morrissey, M. C. Haffner, R. P. Koche, C. L. Sawyers, C. M. Rudin, Exportin 1 inhibition prevents neuroendocrine transformation through SOX2 down-regulation in lung and prostate cancers. Sci Transl Med 15, eadf7006 (2023). 59. A. Quintanal-Villalonga, H. Taniguchi, Y. A. Zhan, M. M. Hasan, S. S. Chavan, F. Meng, F. Uddin, P. Manoj, M. T. A. Donoghue, H. H. Won, J. M. Chan, M. Ciampricotti, A. Chow, M. Offin, J. C. Chang, J. Ray-Kirton, S. E. Tischfield, J. Egger, U. K. Bhanot, I. Linkov, M. Asher, S. Sinha, J. Silber, C. A. Iacobuzio-Donahue, M. H. Roehrl, T. J. Hollmann, H. A. Yu, J. Qiu, E. de Stanchina, M. K. Baine, N. Rekhtman, J. T. Poirier, B. Loomis, R. P. Koche, C. M. Rudin, T. Sen, Multiomic Analysis of Lung Tumors Defines Pathways Activated in Neuroendocrine Transformation. Cancer Discov 11, 3028-3047 (2021). -81- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 60. A. Quintanal-Villalonga, S. Molina-Pinelo, C. Cirauqui, L. Ojeda-Marquez, A. Marrugal, R. Suarez, E. Conde, S. Ponce-Aix, A. B. Enguita, A. Carnero, I. Ferrer, L. Paz- Ares, FGFR1 Cooperates with EGFR in Lung Cancer Oncogenesis, and Their Combined Inhibition Shows Improved Efficacy. J Thorac Oncol 14, 641-655 (2019). 61. A. Dobin, C. A. Davis, F. Schlesinger, J. Drenkow, C. Zaleski, S. Jha, P. Batut, M. Chaisson, T. R. Gingeras, STAR: ultrafast universal RNA-seq aligner. Bioinformatics 29, 15-21 (2013). 62. P. G. Engstrom, T. Steijger, B. Sipos, G. R. Grant, A. Kahles, G. Ratsch, N. Goldman, T. J. Hubbard, J. Harrow, R. Guigo, P. Bertone, R. Consortium, Systematic evaluation of spliced alignment programs for RNA-seq data. Nat Methods 10, 1185-1191 (2013). 63. T. Wu, E. Hu, S. Xu, M. Chen, P. Guo, Z. Dai, T. Feng, L. Zhou, W. Tang, L. Zhan, X. Fu, S. Liu, X. Bo, G. Yu, clusterProfiler 4.0: A universal enrichment tool for interpreting omics data. Innovation (Camb) 2, 100141 (2021). 64. A. Subramanian, P. Tamayo, V. K. Mootha, S. Mukherjee, B. L. Ebert, M. A. Gillette, A. Paulovich, S. L. Pomeroy, T. R. Golub, E. S. Lander, J. P. Mesirov, Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc Natl Acad Sci U S A 102, 15545-15550 (2005). 65. A. Liberzon, A. Subramanian, R. Pinchback, H. Thorvaldsdottir, P. Tamayo, J. P. Mesirov, Molecular signatures database (MSigDB) 3.0. Bioinformatics 27, 1739-1740 (2011). 66. R. Patro, G. Duggal, M. I. Love, R. A. Irizarry, C. Kingsford, Salmon provides fast and bias-aware quantification of transcript expression. Nat Methods 14, 417-419 (2017). 67. D. R. Zerbino, P. Achuthan, W. Akanni, M. R. Amode, D. Barrell, J. Bhai, K. Billis, C. Cummins, A. Gall, C. G. Giron, L. Gil, L. Gordon, L. Haggerty, E. Haskell, T. Hourlier, O. G. Izuogu, S. H. Janacek, T. Juettemann, J. K. To, M. R. Laird, I. Lavidas, Z. Liu, J. E. Loveland, T. Maurel, W. McLaren, B. Moore, J. Mudge, D. N. Murphy, V. Newman, M. Nuhn, D. Ogeh, C. K. Ong, A. Parker, M. Patricio, H. S. Riat, H. Schuilenburg, D. Sheppard, H. Sparrow, K. Taylor, A. Thormann, A. Vullo, B. Walts, A. Zadissa, A. Frankish, S. E. Hunt, M. Kostadima, N. Langridge, F. J. Martin, M. Muffato, E. Perry, M. Ruffier, D. M. Staines, S. J. Trevanion, B. L. Aken, F. Cunningham, A. Yates, P. Flicek, Ensembl 2018. Nucleic Acids Res 46, D754-D761 (2018). -82- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 68. H. Pimentel, N. L. Bray, S. Puente, P. Melsted, L. Pachter, Differential analysis of RNA-seq incorporating quantification uncertainty. Nat Methods 14, 687-690 (2017). 69. G. Yu, L. G. Wang, Y. Han, Q. Y. He, clusterProfiler: an R package for comparing biological themes among gene clusters. OMICS 16, 284-287 (2012). 70. A. Colaprico, T. C. Silva, C. Olsen, L. Garofano, C. Cava, D. Garolini, T. S. Sabedot, T. M. Malta, S. M. Pagnotta, I. Castiglioni, M. Ceccarelli, G. Bontempi, H. Noushmehr, TCGAbiolinks: an R/Bioconductor package for integrative analysis of TCGA data. Nucleic Acids Res 44, e71 (2016). 71. T. C. Silva, A. Colaprico, C. Olsen, F. D'Angelo, G. Bontempi, M. Ceccarelli, H. Noushmehr, TCGA Workflow: Analyze cancer genomics and epigenomics data using Bioconductor packages. F1000Res 5, 1542 (2016). 72. M. Mounir, M. Lucchetta, T. C. Silva, C. Olsen, G. Bontempi, X. Chen, H. Noushmehr, A. Colaprico, E. Papaleo, New functionalities in the TCGAbiolinks package for the study and integration of cancer data from GDC and GTEx. PLoS Comput Biol 15, e1006701 (2019). 73. L. Mangiante, N. Alcala, A. Sexton-Oates, A. Di Genova, A. Gonzalez-Perez, A. Khandekar, E. N. Bergstrom, J. Kim, X. Liu, R. Blazquez-Encinas, C. Giacobi, N. Le Stang, S. Boyault, C. Cuenin, S. Tabone-Eglinger, F. Damiola, C. Voegele, M. Ardin, M. C. Michallet, L. Soudade, T. M. Delhomme, A. Poret, M. Brevet, M. C. Copin, S. Giusiano- Courcambeck, D. Damotte, C. Girard, V. Hofman, P. Hofman, J. Mouroux, C. Cohen, S. Lacomme, J. Mazieres, V. T. de Montpreville, C. Perrin, G. Planchard, N. Rousseau, I. Rouquette, C. Sagan, A. Scherpereel, F. Thivolet, J. M. Vignaud, D. Jean, A. G. S. Ilg, R. Olaso, V. Meyer, A. Boland-Auge, J. F. Deleuze, J. Altmuller, P. Nuernberg, A. Ibanez- Costa, J. P. Castano, S. Lantuejoul, A. Ghantous, C. Maussion, P. Courtiol, H. Hernandez- Vargas, C. Caux, N. Girard, N. Lopez-Bigas, L. B. Alexandrov, F. Galateau-Salle, M. Foll, L. Fernandez-Cuesta, Multiomic analysis of malignant pleural mesothelioma identifies molecular axes and specialized tumor profiles driving intertumor heterogeneity. Nat Genet 55, 607-618 (2023). -83- 4932-3602-9469.1

Claims

Atty. Dkt. No.115872-3182 WHAT IS CLAIMED IS 1. A method for treating diffuse pleural mesothelioma (DPM) in a subject in need thereof comprising administering to the subject an effective amount of an antibody-drug conjugate comprising an anti-TROP-2 antibody or antigen binding fragment thereof. 2. A method for treating diffuse pleural mesothelioma (DPM) in a subject in need thereof comprising administering to the subject an effective amount of an antibody-drug conjugate comprising an anti-TROP-2 antibody or antigen binding fragment thereof and an effective amount of an AKT inhibitor. 3. A method for re-sensitizing DPM tumors to anti-TROP-2 antibody-drug conjugate therapy comprising administering to the subject an effective amount of an antibody-drug conjugate comprising an anti-TROP-2 antibody or antigen binding fragment thereof and an effective amount of an AKT inhibitor. 4. The method of claim 2 or 3, wherein the AKT inhibitor is selected from the group consisting of samotolisib, oridonin, capivasertib, ipatasertib, miransertib, afuresertib, uprosertib, BAY1125976, MK-2206, TAS-117, GSK690693, triciribine, and perifosine, or wherein the AKT inhibitor is administered intravenously, intraperitoneally, subcutaneously, intramuscularly, or intratumorally. 5. The method of any one of claims 2-4, wherein the antibody-drug conjugate and the AKT inhibitor are administered separately, simultaneously, or sequentially. 6. The method of any one of claims 1-5, wherein the antibody-drug conjugate comprises one or more of alkylating agents, topoisomerase inhibitors, platinum agents, taxanes, vinca agents, anti-estrogen drugs, aromatase inhibitors, ovarian suppression agents, VEGF/VEGFR inhibitors, EGF/EGFR inhibitors, PARP inhibitors, cytostatic alkaloids, cytotoxic antibiotics, antimetabolites, endocrine/hormonal agents, and bisphosphonate therapy agents. 7. The method of any one of claims 1-6, wherein the antibody-drug conjugate comprises a cytotoxic drug selected from the group consisting of an anthracycline, a camptothecin, a tubulin inhibitor, a maytansinoid, a calicheamycin, an auristatin, a nitrogen mustard, an ethylenimine derivative, an alkyl sulfonate, a nitrosourea, a triazene, a folic acid analog, a taxane, a COX-2 inhibitor, a pyrimidine analog, a purine analog, an antibiotic, an enzyme inhibitor, an epipodophyllotoxin, a platinum coordination complex, a vinca alkaloid, a substituted urea, a methyl hydrazine derivative, an adrenocortical suppressant, a hormone antagonist, an antimetabolite, an alkylating agent, an antimitotic, an anti- -84- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 angiogenic agent, a tyrosine kinase inhibitor, an mTOR inhibitor, a heat shock protein (HSP90) inhibitor, a proteosome inhibitor, an HDAC inhibitor, a topoisomerase inhibitor and a pro-apoptotic agent. 8. The method of any one of claims 1-7, wherein the antibody-drug conjugate comprises cyclophosphamide, fluorouracil (or 5-fluorouracil or 5-FU), methotrexate, edatrexate (10-ethyl-10-deaza-aminopterin), thiotepa, carboplatin, cisplatin, taxanes, paclitaxel, protein-bound paclitaxel, docetaxel, vinorelbine, tamoxifen, raloxifene, toremifene, fulvestrant, gemcitabine, irinotecan, ixabepilone, temozolmide, topotecan, vincristine, vinblastine, eribulin, mutamycin, capecitabine, anastrozole, exemestane, letrozole, leuprolide, abarelix, buserlin, goserelin, megestrol acetate, risedronate, pamidronate, ibandronate, alendronate, denosumab, zoledronate, trastuzumab, tykerb, anthracyclines, bevacizumab, Tesirine, afatinib, aplidin, azaribine, axitinib, AVL-101, AVL-291, bendamustine, bleomycin, bortezomib, bosutinib, bryostatin-1, busulfan, calicheamycin, camptothecin, 10-hydroxy camptothecin, carmustine, celecoxib, chlorambucil, COX-2 inhibitors, SN-38, cladribine, camptothecans, crizotinib, cytarabine, dacarbazine, dasatinib, dinaciclib, dactinomycin, daunorubicin, deruxtecan, DM1, DM3, DM4, doxorubicin, 2-pyrrolinodoxorubicine (2-PDox), a pro-drug form of 2-PDox (pro-2- PDox), cyano-morpholino doxorubicin, doxorubicin glucuronide, endostatin, epirubicin glucuronide, erlotinib, estramustine, epidophyllotoxin, erlotinib, entinostat, estrogen receptor binding agents, etoposide (VP 16), etoposide glucuronide, etoposide phosphate, fmgolimod, floxuridine (FUdR), 3',5'-O-dioleoyl-FudR (FUdR-dO), fludarabine, flutamide, fame syl -protein transferase inhibitors, flavopiridol, fostamatinib, ganetespib, GDC-0834, GS-1101, gefitinib, hydroxyurea, ibrutinib, idarubicin, idelalisib, ifosfamide, imatinib, lapatinib, lenolidamide, leucovorin, LFM-A13, lomustine, mechlorethamine, melphalan, mercaptopurine, 6-mercaptopurine, mitoxantrone, mithramycin, mitomycin, mitotane, monomethylauristatin F (MMAF), monomethylauristatin D (MMAD), monomethylauristatin E (MMAE), navelbine, neratinib, nilotinib, nitrosurea, olaparib, plicomycin, procarbazine, PCI-32765, pentostatin, PSI-341, semustine, SN-38, sorafenib, streptozocin, SU11248, sunitinib, transplatinum, thalidomide, thioguanine, teniposide, uracil mustard, vatalanib, vinca alkaloids, ZD1839 or combinations thereof. 9. The method of any one of claims 1-8, wherein the anti-TROP-2 antibody or antigen binding fragment comprises the variable heavy (VH) domain and variable light (VL) domain of sacituzumab. -85- 4932-3602-9469.1 Atty. Dkt. No.115872-3182 10. The method of claim 9, wherein the VH domain comprises a VH-CDR1 sequence of SEQ ID NO: 5, a VH-CDR2 sequence of SEQ ID NO: 6, and a VH-CDR3 sequence of SEQ ID NO: 7 and the VL domain comprises a VL-CDR1 sequence of SEQ ID NO: 8, a VL- CDR2 sequence of SEQ ID NO: 9, and a VL-CDR3 sequence of SEQ ID NO: 10. 11. The method of claim 9 or 10, wherein (a) the VH domain comprises an amino acid sequence of SEQ ID NO: 3; and (b) the VL domain comprises an amino acid sequence of SEQ ID NO: 1. 12. The method of any one of claims 1-8, wherein the anti-TROP-2 antibody or antigen binding fragment comprises the variable heavy domain and variable light domain of datopotamab. 13. The method of claim 12, wherein the VH domain comprises a VH-CDR1 sequence of SEQ ID NO: 15, a VH-CDR2 sequence of SEQ ID NO: 16, and a VH-CDR3 sequence of SEQ ID NO: 17 and the VL domain comprises a VL-CDR1 sequence of SEQ ID NO: 18, a VL-CDR2 sequence of SEQ ID NO: 19, and a VL-CDR3 sequence of SEQ ID NO: 20. 14. The method of claim 12 or 13, wherein: (a) the VH domain comprises an amino acid sequence of SEQ ID NO: 13; and (b) the VL domain comprises an amino acid sequence of SEQ ID NO: 11. 15. The method of any one of claims 1-14, wherein the subject has received a prior anti- cancer therapy. 16. The method of any one of claims 1-15, wherein the subject is diagnosed with or is suffering from recurrent DPM or metastatic DPM. 17. The method of any one of claims 1-11 or 15-16, wherein the antibody-drug conjugate comprises sacituzumab govitecan. 18. The method of any one of claims 1-17, wherein the antibody-drug conjugate is administered intravenously, intraperitoneally, subcutaneously, intramuscularly, or intratumorally. -86- 4932-3602-9469.1
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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20230101735A1 (en) * 2020-01-22 2023-03-30 Jiangsu Hengrui Medicine Co., Ltd. Anti-trop-2 antidody-exatecan analog conjugate and medical use thereof
WO2024030758A1 (en) * 2022-08-01 2024-02-08 Iovance Biotherapeutics, Inc. Chimeric costimulatory receptors, chemokine receptors, and the use of same in cellular immunotherapies

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20230101735A1 (en) * 2020-01-22 2023-03-30 Jiangsu Hengrui Medicine Co., Ltd. Anti-trop-2 antidody-exatecan analog conjugate and medical use thereof
WO2024030758A1 (en) * 2022-08-01 2024-02-08 Iovance Biotherapeutics, Inc. Chimeric costimulatory receptors, chemokine receptors, and the use of same in cellular immunotherapies

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