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WO2024206274A2 - T cell receptors targeting y220c or r175h mutation in p53 - Google Patents

T cell receptors targeting y220c or r175h mutation in p53 Download PDF

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Publication number
WO2024206274A2
WO2024206274A2 PCT/US2024/021425 US2024021425W WO2024206274A2 WO 2024206274 A2 WO2024206274 A2 WO 2024206274A2 US 2024021425 W US2024021425 W US 2024021425W WO 2024206274 A2 WO2024206274 A2 WO 2024206274A2
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WIPO (PCT)
Prior art keywords
seq
amino acid
polypeptide chain
acid sequence
chain comprises
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PCT/US2024/021425
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French (fr)
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WO2024206274A3 (en
Inventor
Sanghyun Kim
Nikolaos ZACHARAKIS
Noam LEVIN
Steven A. Rosenberg
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US Department of Health and Human Services
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US Department of Health and Human Services
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Priority to AU2024241888A priority Critical patent/AU2024241888A1/en
Priority to CN202480029232.1A priority patent/CN121127261A/en
Publication of WO2024206274A2 publication Critical patent/WO2024206274A2/en
Publication of WO2024206274A3 publication Critical patent/WO2024206274A3/en
Anticipated expiration legal-status Critical
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70503—Immunoglobulin superfamily
    • C07K14/7051—T-cell receptor (TcR)-CD3 complex
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K40/00—Cellular immunotherapy
    • A61K40/10—Cellular immunotherapy characterised by the cell type used
    • A61K40/11—T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K40/00—Cellular immunotherapy
    • A61K40/30—Cellular immunotherapy characterised by the recombinant expression of specific molecules in the cells of the immune system
    • A61K40/32—T-cell receptors [TCR]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K40/00—Cellular immunotherapy
    • A61K40/40—Cellular immunotherapy characterised by antigens that are targeted or presented by cells of the immune system
    • A61K40/41—Vertebrate antigens
    • A61K40/42—Cancer antigens
    • A61K40/4238—Regulators of development
    • A61K40/424—Apoptosis related proteins, e.g. survivin or livin
    • A61K40/4241—Apoptosis related proteins, e.g. survivin or livin p53
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
    • C07K14/4746—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used p53

Definitions

  • TCR T cell receptor
  • Leydig 770310 HHS E-054-2023-0-PC-01 2 Further aspects of the invention polypeptides, proteins, nucleic acids, recombinant expression vectors, host cells, populations of cells, and pharmaceutical compositions relating to the TCRs of the invention. Still further aspects of the invention provide methods of detecting the presence of cancer in a mammal, methods of inducing an immune response against a cancer in a mammal, and methods of treating or preventing cancer in a mammal. Additional aspects of the invention provide methods of producing a host cell expressing the TCR and methods of producing the TCR, polypeptide, or protein.
  • Figure 1A is a graph showing the frequency (%) of tetramer positive cells measured following each of culture conditions 1, 2, and 3 of Table 1. Tetramers were generated from HLA-A*02:01 monomer and were loaded with p53 R175H 9-mer peptide analogue to capture p53 R175H mutation-reactive T cells with HLA-A*02:01 restriction.
  • Figures 1B-1E are graphs showing the percentage of mTCR+CD8+ cells expressing 4-1BB following co-culture of effector cells with target cells.
  • Effector cells were healthy donor PBL transduced with 4196-C TCR (B), 4196-D TCR (C), 4196-E TCR (D) or TCR 4196_AV6_with_BV11-2 (E) (disclosed in US 2020/0277352; used as the “Comparison TCR”).
  • Target cells were T2 cells that naturally expressed HLA-A2 and had been incubated with the WT peptide analogue or mutant p53 R175H 9-mer peptide analogue at the indicated concentrations.
  • Figure 1F is a graph including the same data shown in Figures 1B-1E replotted together for comparison.
  • Figure 1G is a graph showing the normalized tumor cell counts measured at 4 hour intervals following co-culture of effector cells with target cells.
  • Effector cells were healthy donor PBL transduced with a retroviral vector encoding 4196-C TCR, 4196-D TCR, 4196-E TCR, or an irrelevant TCR.
  • the target cells were TYK-nu human ovarian cancer cells that naturally expressed p53 R175H and HLA-A*02:01. P ⁇ 0.001.
  • Figure 2A shows flow cytometry data obtained after staining TILs from Patient 4424 for CD8 and tetramer, which was used in Figure 1A.
  • Figure 2B is a graph showing the percentage of mTCR+CD8+ cells expressing 4- 1BB as a marker for T cell activation following co-culture of effector cells with target cells.
  • Effector cells were healthy donor PBL transduced with the 4424-R175H TCR.
  • Target cells Leydig 770310 HHS E-054-2023-0-PC-01 3 were monkey kidney COS7 cells engineered express HLA-A2 and incubated with the WT peptide analogue or mutant p53 R175H 9-mer peptide analogue at the indicated concentrations.
  • Figure 3 is a graph showing the normalized tumor cell counts measured at 3 hour intervals following co-culture of effector cells with target cells at an effector to target ratio of 2:1.
  • Effector cells were healthy donor PBL transduced with a retroviral vector encoding the 4196-C TCR, 4196-D TCR, 4196-E TCR, 4424-R175H TCR, or 4141-TCR1a2 disclosed in US 2022/0332785 (another “Comparison TCR”).
  • the target cells were TYK-nu human ovarian cancer cells.
  • Figures 4A-4D are graphs showing the percentage of mTCR+CD3+ cells expressing 4-1BB following co-culture of effector cells with target cells.
  • Effector cells were healthy donor PBL transduced with the 4402-Y220C-B TCR, 4402-Y220C-C TCR, or 4402- Y220C-L TCR or the 4343-D TCR (“Comparison TCR”).
  • Target cells were autologous B cells pulsed with WT or mutant peptide at the indicated concentrations.
  • Figure 4E is a graph showing the percentage of mTCR+CD3+ cells expressing 4- 1BB following co-culture of effector cells with target cells. Effector cells were healthy donor PBL transduced with the 4402-Y220C-C TCR or 4402-Y220C-L TCR.
  • Target cells were autologous breast tumor organoid cells from Patient 4402 of passage 11 showing loss of DRB1*13:03 or passage 27 showing reduced but detectable levels of DRB1*13:03 expression (see Figure 4F).
  • TCR-C and L show recognition of only the organoid passage 27 but not the passage 11, suggesting the two TCRs are restricted by DRB1*13:03.
  • Target cells pulsed with DMSO served as a control.
  • Figure 4F is a graph showing the Transcripts Per Kilobase Million (TPM) value measured for HLA-DRB1*15:01 and DRB1*13:03 in the autologous organoid cells from Patient 4402 of passages 11 and 27.
  • TPM Transcripts Per Kilobase Million
  • Figures 5A-5C show an alignment of the amino acid sequences of the nine p53 splice variants.
  • P53_HUMAN SEQ ID NO: 1
  • P53_HUMAN SEQ ID NO: 104
  • P53_HUMAN SEQ ID NO: 105
  • P53_HUMAN SEQ ID NO: 106
  • P53_HUMAN SEQ ID NO: 107
  • P53_HUMAN SEQ ID NO: 111).
  • FIGS 6-7 are graphs showing mean tumor size (mm 2 ) measured at the indicated number of days after adoptive cell transfer (ACT) of transduced healthy donor T cells administered to tumor-bearing mice at a dose of 2 million cells/mouse ( Figure 6) or 10 million cells/mouse ( Figure 7).
  • ACT adoptive cell transfer
  • the healthy donor T cells were independently transduced with: (i) the retroviral vector encoding the 4196-C TCR of Example 4 (“4196 IVS-C”); (ii) an irrelevant TCR control (i.e., 4259) (“4259 (irrelevant)”); (iii) the TCR 4196_AV12- 1_with_BV6-1 disclosed in US 2020/0277352 (“12-6”); (iv) the TCR 4196_AV38- 1_with_BV10-3 disclosed in US 2020/0277352 (“38-10”); (v) the TCR 4196_AV6_with_BV11-2 disclosed in US 2020/0277352 (“6-11”); (vi) the TCR 4141- TCR1a2 disclosed in US 2022/0332785 (“41411a2”); or (vii) the 4141 IVS TCR disclosed in US Patent Application No.18/289,596 (“4141 IVS”).
  • Tumor Protein P53 acts as a tumor suppressor by, for example, regulating cell division.
  • the p53 protein is located in the nucleus of the cell, where it binds directly to DNA.
  • the p53 protein is involved in determining whether the DNA will be repaired or the damaged cell will undergo apoptosis. If the DNA can be repaired, p53 activates other genes to fix the damage. If the DNA cannot be repaired, the p53 protein prevents the cell from dividing and signals it to undergo apoptosis. By stopping cells with mutated or damaged DNA from dividing, p53 helps prevent the development of tumors.
  • WT (normal) full-length p53 comprises the amino acid sequence of SEQ ID NO: 1. Mutations in the p53 protein may reduce or eliminate the p53 protein’s tumor suppressor function. Alternatively or additionally, a p53 mutation may be a gain-of-function mutation by interfering with WT p53 in a dominant negative fashion. Mutated p53 protein may be expressed in any of a variety of human cancers such as, for example, cholangiocarcinoma, melanoma, colon cancer, rectal cancer, ovarian cancer, endometrial cancer, non-small cell lung cancer (NSCLC), glioblastoma, uterine cervical cancer, head and neck cancer, breast cancer, pancreatic cancer, or bladder cancer.
  • human cancers such as, for example, cholangiocarcinoma, melanoma, colon cancer, rectal cancer, ovarian cancer, endometrial cancer, non-small cell lung cancer (NSCLC), glioblastoma, uterine cervical cancer,
  • An aspect of the invention provides an isolated or purified T cell receptor (TCR) having antigenic specificity for a human p53 Y220C or human p53 R175H amino acid sequence Leydig 770310 HHS E-054-2023-0-PC-01 5 (hereinafter, “mutated p53”).
  • TCR T cell receptor
  • a p53 amino acid sequence may comprise fewer than all of the amino acid residues of the full-length, WT p53 protein. Accordingly, the position numbers are defined herein by reference to the WT full-length p53 protein (namely, SEQ ID NO: 1) with the understanding that the actual position of the corresponding residue in a particular example of a p53 amino acid sequence may be different. Because the positions are as defined by SEQ ID NO: 1, the term “Y220C” indicates that the tyrosine present at position 220 of SEQ ID NO: 1 is replaced by cysteine, and “R175H” indicates that the arginine present at position 175 of SEQ ID NO: 1 has been replaced with histidine.
  • Y220C refers to a substitution of the underlined tyrosine in SEQ ID NO: 114 with cysteine, even though the actual position of the underlined tyrosine in SEQ ID NO: 114 is 13.
  • Human p53 amino acid sequences with the Y220C mutation are hereinafter referred to as “Y220C” or “p53 Y220C .”
  • Human p53 amino acid sequences with the R175H mutation are hereinafter referred to as “R175H” or “p53 R175H .”
  • “mutated p53” refers to human p53 Y220C or human p53 R175H .
  • P53 has nine known splice variants. The p53 mutations described herein are conserved over all nine p53 splice variants. An alignment of the nine p53 splice variants is shown in Figures 5A-5C.
  • the inventive TCRs may have antigenic specificity for any mutated p53 amino acid sequence described herein encoded by any of the nine p53 splice variants. Because the positions are as defined by SEQ ID NO: 1, then the actual positions of the amino acid sequence of a particular splice variant of p53 are defined relative to the corresponding positions of SEQ ID NO: 1, and the positions as defined by SEQ ID NO: 1 may be different than the actual positions in a particular splice variant.
  • mutations refer to a replacement of an amino acid residue in the amino acid sequence of a particular splice variant of p53 corresponding to the indicated position of the 393-amino acid Leydig 770310 HHS E-054-2023-0-PC-01 6 sequence of SEQ ID NO: 1 with the that the actual positions in the splice variant may be different.
  • the TCR has antigenic specificity for human p53 with a mutation at position 220, as defined by SEQ ID NO: 1.
  • the p53 mutation at position 220 may be a missense mutation.
  • the mutation at position 220 may be a substitution of the native (WT) tyrosine residue present at position 220 with any amino acid residue other than tyrosine.
  • the TCR has antigenic specificity for a human p53 Y220C amino acid sequence.
  • the TCR may have antigenic specificity for the human p53 Y220C amino acid sequence of DRNTFRHSVVVPCEPPEVGSDCTTI (SEQ ID NO: 114).
  • the TCR does not have antigenic specificity for the wild-type human p53 amino acid sequence of DRNTFRHSVVVPYEPPEVGSDCTTI (SEQ ID NO: 115).
  • the TCR has antigenic specificity for human p53 with a mutation at position 175, as defined by SEQ ID NO: 1.
  • the p53 mutation at position 175 may be a missense mutation. Accordingly, the mutation at position 175 may be a substitution of the native (WT) arginine residue present at position 175 with any amino acid residue other than arginine.
  • the TCR has antigenic specificity for a human p53 R175H amino acid sequence.
  • the TCR may have antigenic specificity for the human p53 R175H amino acid sequence of HMTEVVRHC (SEQ ID NO: 112).
  • the TCR does not have antigenic specificity for the wild- type human p53 amino acid sequence of HMTEVVRRC (SEQ ID NO: 113).
  • the inventive TCRs may be able to recognize mutated p53 in an HLA (human leukocyte antigen)-molecule-dependent manner.
  • HLA- molecule-dependent manner means that the TCR elicits an immune response upon binding to mutated p53 within the context of an HLA molecule, which HLA molecule is expressed by the patient from which the TCR was isolated.
  • the inventive TCRs may be able to recognize mutated p53 that is presented by the applicable HLA molecule and may bind to the HLA molecule in addition to mutated p53.
  • the inventive TCRs are able to recognize Y220C presented by an HLA Class II molecule.
  • the TCR may elicit an immune response upon binding to Y220C within the context of an HLA Class II molecule.
  • the inventive TCRs are able to recognize Y220C that is presented by an HLA Class II molecule and may bind to the HLA Class II molecule in addition to Y220C.
  • the Class II molecule is an HLA-DR heterodimer.
  • the HLA-DR heterodimer is a cell surface receptor including an ⁇ chain and a ⁇ chain.
  • the HLA-DR ⁇ chain is encoded by the HLA-DRA gene.
  • the HLA-DR ⁇ chain is encoded by the HLA-DRB1 gene, the HLA-DRB3 gene, HLA-DRB4 gene, or the HLA- DRB5 gene.
  • Examples of molecules encoded by the HLA-DRB1 gene may include, but are not limited to, HLA-DR1, HLA-DR2, HLA-DR3, HLA-DR4, HLA-DR5, HLA-DR6, HLA- DR7, HLA-DR8, HLA-DR9, HLA-DR10, HLA-DR11, HLA-DR12, HLA-DR13, HLA- DR14, HLA-DR15, HLA-DR16, and HLA-DR17.
  • the HLA-DRB3 gene encodes HLA- DR52.
  • the HLA-DRB4 gene encodes HLA-DR53.
  • the HLA-DRB5 gene encodes HLA- DR51.
  • the alpha chain of the HLA Class II molecule is expressed by the HLA-DRA1*01:01 allele.
  • the beta chain of the HLA Class II molecule is expressed by the HLA-DRB3*02:02 allele.
  • the HLA Class II molecule is an HLA-DRB3:HLA-DRA heterodimer.
  • the HLA Class II molecule is a heterodimer of an HLA-DRA1*01:01 chain and an HLA-DRB3*02:02 chain.
  • the mutated p53 is Y220C and the HLA Class II molecule is a heterodimer of an HLA-DRA1*01:01 chain and an HLA-DRB3*02:02 chain.
  • the alpha chain of the HLA Class II molecule is expressed by the HLA-DRA1*01:01 allele.
  • the beta chain of the HLA Class II molecule is expressed by the HLA-DRB1*13:03 allele.
  • the HLA Class II molecule is an HLA-DRB1:HLA-DRA heterodimer.
  • the HLA Class II molecule is a heterodimer of an HLA-DRA1*01:01 chain and an HLA-DRB1*13:03 chain.
  • the mutated p53 is Y220C and the HLA Class II molecule is a heterodimer of an HLA-DRA1*01:01 chain and an HLA-DRB1*13:03 chain.
  • the inventive TCRs are able to recognize R175H presented by an HLA Class I molecule. In this regard, the TCR may elicit an immune response upon binding to R175H within the context of an HLA Class I molecule.
  • the inventive TCRs are able to recognize R175H that is presented by an HLA Class I molecule and may bind to the HLA Class I molecule in addition to R175H.
  • the HLA Class I molecule is an HLA-A molecule.
  • the HLA-A molecule is a heterodimer of an ⁇ chain and ⁇ 2 microglobulin.
  • the HLA-A ⁇ chain may be encoded by an HLA-A gene.
  • ⁇ 2 microglobulin binds non-covalently to the alpha1, alpha2 and alpha3 domains of the alpha chain to build the HLA-A complex.
  • the HLA-A molecule may be any HLA-A
  • the HLA Class I molecule is an HLA-A2 molecule.
  • the HLA-A2 molecule may be any HLA- A2 molecule.
  • Examples of HLA-A2 molecules may include, but are not limited to, those encoded by the HLA-A*02:01, HLA-A*02:02, HLA-A*02:03 allele, HLA-A*02:05, HLA- A*02:06, HLA-A*02:07 allele, or HLA-A*02:11 allele.
  • the HLA Class I molecule is encoded by the HLA-A*02:01 allele.
  • the TCRs of the invention may provide any one or more of many advantages, including when expressed by cells used for adoptive cell transfer. Mutated p53 is expressed by cancer cells and is not expressed by normal, noncancerous cells. Without being bound to a particular theory or mechanism, it is believed that the inventive TCRs advantageously target the destruction of cancer cells while minimizing or eliminating the destruction of normal, non-cancerous cells, thereby reducing, for example, by minimizing or eliminating, toxicity. Moreover, the inventive TCRs may, advantageously, successfully treat or prevent mutated p53-positive cancers that do not respond to other types of treatment such as, for example, chemotherapy, surgery, or radiation.
  • the inventive TCRs may provide highly avid recognition of mutated p53, which may provide the ability to recognize unmanipulated tumor cells (e.g., tumor cells that have not been treated with interferon (IFN)- ⁇ , transfected with a vector encoding one or both of mutated p53 and the applicable HLA molecule, pulsed with a p53 peptide with the p53 mutation, or a combination thereof).
  • unmanipulated tumor cells e.g., tumor cells that have not been treated with interferon (IFN)- ⁇ , transfected with a vector encoding one or both of mutated p53 and the applicable HLA molecule, pulsed with a p53 peptide with the p53 mutation, or a combination thereof.
  • Mutations in p53 are common across different tumor types. Roughly half of all tumors harbor a mutation in p53, about half of which will be a missense mutation. The R175H mutation is common, affecting about 5% of all patients with solid cancers.
  • the Y220C mutation is also highly recurrent and affects about 0.4% of all cancer patients. Accordingly, the inventive TCRs may increase the number of patients who may be eligible for treatment with immunotherapy.
  • the phrase “antigenic specificity,” as used herein, means that the TCR can specifically bind to and immunologically recognize mutated p53 with high avidity.
  • a TCR may be considered to have “antigenic specificity” for mutated p53 if about 1 x 10 4 to about 1 x 10 5 T cells expressing the TCR secrete at least about 200 pg/mL or more (e.g., 200 pg/mL or more, 300 pg/mL or more, 400 pg/mL or more, 500 pg/mL or more, 600 pg/mL or more, 700 pg/mL or more, 1000 pg/mL or more, 5,000 pg/mL or more, 7,000 pg/mL or more, 10,000 pg/mL or more, 20,000 pg/mL or more, or a range defined by any two of the foregoing values) of IFN- ⁇ upon co-culture with (a) antigen-negative, applicable HLA molecule positive target cells pulsed with mutated p53 peptide (e.g., about 0.1 ng/mL to Leydig 770310 H
  • TCRs expressing the inventive TCRs may also secrete IFN- ⁇ upon co-culture with antigen-negative, applicable HLA molecule positive target cells pulsed with higher concentrations of mutated p53 peptide.
  • a TCR may be considered to have “antigenic specificity” for mutated p53 if T cells expressing the TCR secrete at least twice as much IFN- ⁇ upon co-culture with (a) antigen-negative, applicable HLA molecule positive target cells pulsed with mutated p53 peptide or (b) antigen-negative, applicable HLA molecule positive target cells into which a nucleotide sequence encoding mutated p53 has been introduced such that the target cell expresses mutated p53 as compared to the amount of IFN- ⁇ expressed by a negative control.
  • the negative control may be, for example, (i) T cells expressing the TCR, co-cultured with (a) antigen-negative, applicable HLA molecule positive target cells pulsed with the same concentration of an irrelevant peptide (e.g., some other peptide with a different sequence from the mutated p53 peptide) or (b) antigen-negative, applicable HLA molecule positive target cells into which a nucleotide sequence encoding an irrelevant peptide has been introduced such that the target cell expresses the irrelevant peptide, or (ii) untransduced T cells (e.g., derived from PBMC, which do not express the TCR) co-cultured with (a) antigen- negative, applicable HLA molecule positive target cells pulsed with the same concentration of mutated p53 peptide or (b) antigen-negative, applicable HLA molecule positive target cells into which a nucleotide sequence encoding mutated p53 has been introduced such that the target cell expresses mutated p
  • IFN- ⁇ secretion may be measured by methods known in the art such as, for example, enzyme-linked immunosorbent assay (ELISA).
  • concentration of pulsed peptide may be as described herein with respect to other aspects of the invention.
  • a TCR may be considered to have “antigenic specificity” for mutated p53 if at least twice as many of the numbers of T cells expressing the TCR secrete IFN- ⁇ upon co-culture with (a) antigen-negative, applicable HLA molecule positive target cells pulsed with mutated p53 peptide or (b) antigen-negative, applicable HLA molecule positive target cells into which a nucleotide sequence encoding mutated p53 has been introduced such that the target cell expresses mutated p53 as compared to the numbers Leydig 770310 HHS E-054-2023-0-PC-01 10 of negative control T cells that secrete ⁇ .
  • the concentration of peptide and the negative control may be as described herein with respect to other aspects of the invention.
  • the numbers of cells secreting IFN- ⁇ may be measured by methods known in the art such as, for example, enzyme-linked immunospot (ELISPOT) assay.
  • ELISPOT enzyme-linked immunospot
  • a TCR may be considered to have “antigenic specificity” for mutated p53 if at least twice as many spots are detected by ELISPOT for the T cells expressing the TCR upon co-culture with (a) antigen-negative, applicable HLA molecule positive target cells pulsed with mutated p53 peptide or (b) antigen-negative, applicable HLA molecule positive target cells into which a nucleotide sequence encoding mutated p53 has been introduced such that the target cell expresses mutated p53 as compared to the number of spots detected by ELISPOT for negative control T cells co-cultured with the same target cells.
  • concentration of peptide and the negative control may be as described herein with respect to other aspects of the invention.
  • a TCR may be considered to have “antigenic specificity” for mutated p53 if T cells expressing the TCR upregulate expression of one or both of 4-1BB and OX40 as measured by, for example, flow cytometry after stimulation with target cells expressing mutated p53.
  • An aspect of the invention provides a TCR comprising two polypeptides (i.e., polypeptide chains), such as an alpha ( ⁇ ) chain of a TCR, a beta ( ⁇ ) chain of a TCR, a gamma ( ⁇ ) chain of a TCR, a delta ( ⁇ ) chain of a TCR, or a combination thereof.
  • the polypeptides of the inventive TCR can comprise any amino acid sequence, provided that the TCR has antigenic specificity for mutated p53.
  • the TCR comprises two polypeptide chains, each of which comprises a variable region comprising a complementarity determining region (CDR)1, a CDR2, and a CDR3 of a TCR.
  • CDR complementarity determining region
  • the TCR comprises a first polypeptide chain comprising an ⁇ chain CDR1 (CDR1 ⁇ ), an ⁇ chain CDR2 (CDR2 ⁇ ), and an ⁇ chain CDR3 (CDR3 ⁇ ), and a second polypeptide chain comprising a ⁇ chain CDR1 (CDR1 ⁇ ), a ⁇ chain CDR2 (CDR2 ⁇ ), and a ⁇ chain CDR3 (CDR3 ⁇ ).
  • the TCR comprises the amino acid sequences of: (1) all of SEQ ID NOs: 2-7 (4196-C TCR); (2) all of SEQ ID NOs: 16-21 (4196-D TCR); (3) all of SEQ ID NOs: 30-35 (4196-E TCR); (4) all of SEQ ID NOs: 44-49 (4424-R175H TCR); (5) all of SEQ ID NOs: 58-63 (4402-Y220C-B TCR); (6) all of SEQ ID NOs: 72-77 (4402-Y220C-C TCR); or (7) all of SEQ ID NOs: 86-91 (4402-Y220C-L TCR).
  • the six amino acid sequences in each collection correspond to the CDR1 ⁇ , CDR2 ⁇ , CDR3 ⁇ , CDR1 ⁇ , CDR2 ⁇ , and CDR3 ⁇ of a TCR, respectively.
  • the TCR may comprise the amino acid sequences of any one or more of: SEQ ID NOs: 2-7, 16-21, 30-35, 44-49, 58-63, 72-77, and 86-91.
  • the TCR comprises an isolated or purified T cell receptor (TCR) having antigenic specificity for a human p53 Y220C or human p53 R175H amino acid sequence, wherein the TCR comprises the amino acid sequences of: (1) all of SEQ ID NOs: 2-7; (2) all of SEQ ID NOs: 16-21; (3) all of SEQ ID NOs: 30-35; (4) all of SEQ ID NOs: 44-49; (5) all of SEQ ID NOs: 58-63; (6) all of SEQ ID NOs: 72-77; or (7) all of SEQ ID NOs: 86-91.
  • TCR T cell receptor
  • the TCR comprises an ⁇ chain variable region amino acid sequence and a ⁇ chain variable region amino acid sequence, which together comprise one of the collections of CDRs set forth above.
  • the TCR can comprise the amino acid sequences of: (1) both of SEQ ID NOs: 8 and 9 (4196-C TCR); (2) both of SEQ ID NOs: 10 and 11 (4196-C TCR); (3) both of SEQ ID NOs: 22 and 23 (4196-D TCR); (4) both of SEQ ID NOs: 24 and 25 (4196-D TCR); (5) both of SEQ ID NOs: 36 and 37 (4196-E TCR); (6) both of SEQ ID NOs: 38 and 39 (4196-E TCR); (7) both of SEQ ID NOs: 50 and 51 (4424-R175H TCR); (8) both of SEQ ID NOs: 52 and 53 (4424-R175H TCR); (9) both of SEQ ID NOs: 64 and 65 (4402-Y220C-B TCR); (10)
  • Each one of the foregoing collections of amino acid sequences in this paragraph sets forth the two variable regions of each of seven different TCRs having antigenic specificity for mutated human p53.
  • the two amino acid sequences in each collection correspond to the variable region of the ⁇ chain and the variable region of the ⁇ chain of a TCR, respectively.
  • the TCR may, e.g., comprise the amino acid sequence of any one or more of SEQ ID NOs: 8, 9, 10, 11, 22, 23, 24, 25, 36, 37, 38, 39, 50, 51, 52, 53, 64, 65, 66, 67, 78, 79, 80, 81, 92, 93, 94, and 95.
  • the TCR comprises the amino acid sequence(s) of: (1) SEQ ID NO: 8; (2) SEQ ID NO: 9; (3) both of SEQ ID NOs: 8 and 9; (4) SEQ ID NO: 10; (5) SEQ ID NO: 11; (6) both of SEQ ID NOs: 10 and 11; (7) SEQ ID NO: 22; (8) SEQ ID NO: 23; (9) both of SEQ ID NOs: 22 and 23; (10) SEQ ID NO: 24; (11) SEQ Leydig 770310 HHS E-054-2023-0-PC-01 12 ID NO: 25; (12) both of SEQ ID NOs: 24 (13) SEQ ID NO: 36; (14) SEQ ID NOs: 37; (15) both of SEQ ID NOs: 36 and 37; (16) SEQ ID NO: 38; (17) SEQ ID NO: 39; (18) both of SEQ ID NOs: 38 and 39; (19) SEQ ID NO: 50; (20) SEQ ID NO: 51; (21) both of SEQ ID NOs: 50; (20) SEQ ID
  • the inventive TCRs may further comprise a constant region.
  • the constant region may be derived from any suitable species such as, e.g., human or mouse.
  • the TCRs further comprise a murine constant region.
  • murine or “human,” when referring to a TCR or any component of a TCR described herein (e.g., complementarity determining region (CDR), variable region, constant region, alpha chain, and/or beta chain), means a TCR (or component thereof) which is derived from a mouse or a human, respectively, i.e., a TCR (or component thereof) that originated from or was, at one time, expressed by a mouse T cell or a human T cell, respectively.
  • CDR complementarity determining region
  • the TCR may comprise a murine ⁇ chain constant region and a murine ⁇ chain constant region.
  • the murine ⁇ chain constant region may be modified or unmodified.
  • a modified murine ⁇ chain constant region may be, e.g., cysteine-substituted, LVL-modified, or both cysteine-substituted and LVL-modified, as described, for example, in U.S. Patent No. 10,174,098.
  • the murine ⁇ chain constant region may be modified or unmodified.
  • a modified murine ⁇ chain constant region may be, e.g., cysteine-substituted, as described, for example, in U.S. Patent No.10,174,098.
  • the TCR comprises a cysteine-substituted, LVL-modified murine ⁇ chain constant region comprising the amino acid sequence of SEQ ID NO: 100 or 101.
  • the TCR comprises a cysteine-substituted murine ⁇ chain constant region comprising the amino acid sequence of SEQ ID NO: 102.
  • the inventive TCR can comprise an ⁇ chain of a TCR and a ⁇ chain of a TCR.
  • the ⁇ chain of the TCR may comprise a variable region of an ⁇ chain and a constant region of an ⁇ chain.
  • An ⁇ chain of this type can be paired with any ⁇ Leydig 770310 HHS E-054-2023-0-PC-01 13 chain of a TCR.
  • the ⁇ chain may a region of a ⁇ chain and a constant region of a ⁇ chain.
  • the TCR can comprise the amino acid sequences of: (1) both of SEQ ID NOs: 12 and 13 (4196-C TCR); (2) both of SEQ ID NOs: 14 and 15 (4196-C TCR); (3) both of SEQ ID NOs: 26 and 27 (4196-D TCR); (4) both of SEQ ID NOs: 28 and 29 (4196-D TCR); (5) both of SEQ ID NOs: 40 and 41 (4196-E TCR); (6) both of SEQ ID NOs: 42 and 43 (4196-E TCR); (7) both of SEQ ID NOs: 54 and 55 (4424-R175H TCR); (8) both of SEQ ID NOs: 56 and 57 (4424-R175H); (9) both of SEQ ID NOs: 68 and 69 (4402-Y220C-B TCR); (10) both of SEQ ID NOs: 70 and 71 (4402-Y220C-B TCR); (11) SEQ ID NO: 82 and 83 (4402-Y220C
  • the TCR may comprise the amino acid sequence of any one or more of SEQ ID NOs: 12, 13, 14, 15, 26, 27, 28, 29, 40, 41, 42, 43, 54, 55, 56, 57, 68, 69, 70, 71, 82, 83, 84, 85, 96, 97, 98, and 99.
  • the TCR comprises the amino acid sequences of: (1) SEQ ID NO: 12; (2) SEQ ID NO: 13; (3) both of SEQ ID NOs: 12 and 13; (4) SEQ ID NO: 14; (5) SEQ ID NO: 15; (6) both of SEQ ID NOs: 14 and 15; (7) SEQ ID NO: 26; (8) SEQ ID NO: 27; (9) both of SEQ ID NOs: 26 and 27; (10) SEQ ID NO: 28; (11) SEQ ID NO: 29; (12) both of SEQ ID NOs: 28 and 29; (13) SEQ ID NO: 40; (14) SEQ ID NO: 41; (15) both of SEQ ID NOs: 40 and 41; (16) SEQ ID NO: 42; (17) SEQ ID NO: 43; (18) both of SEQ ID NOs: 42 and 43; (19) SEQ ID NO: 54; (20) SEQ ID NO: 55; (21) both of SEQ ID NOs: 54 and 55; (22) SEQ ID NO: 56; (23) SEQ ID NO:
  • the term “functional variant,” as used herein, refers to a TCR, Leydig 770310 HHS E-054-2023-0-PC-01 14 polypeptide, or protein having substantial or sequence identity or similarity to a parent TCR, polypeptide, or protein, which functional variant retains the biological activity of the TCR, polypeptide, or protein of which it is a variant.
  • Functional variants encompass, for example, those variants of the TCR, polypeptide, or protein described herein (the parent TCR, polypeptide, or protein) that retain the ability to specifically bind to mutated p53 for which the parent TCR has antigenic specificity or to which the parent polypeptide or protein specifically binds, to a similar extent, the same extent, or to a higher extent, as the parent TCR, polypeptide, or protein.
  • the functional variant can, for instance, be at least about 30%, at least about 50%, at least about 75%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or more identical in amino acid sequence to the parent TCR, polypeptide, or protein, respectively.
  • the functional variant can, for example, comprise the amino acid sequence of the parent TCR, polypeptide, or protein with at least one conservative amino acid substitution. Conservative amino acid substitutions are known in the art, and include amino acid substitutions in which one amino acid having certain physical and/or chemical properties is exchanged for another amino acid that has the same chemical or physical properties.
  • the conservative amino acid substitution can be an acidic amino acid substituted for another acidic amino acid (e.g., Asp or Glu), an amino acid with a nonpolar side chain substituted for another amino acid with a nonpolar side chain (e.g., Ala, Gly, Val, Ile, Leu, Met, Phe, Pro, Trp, Val, etc.), a basic amino acid substituted for another basic amino acid (Lys, Arg, etc.), an amino acid with a polar side chain substituted for another amino acid with a polar side chain (Asn, Cys, Gln, Ser, Thr, Tyr, etc.), etc.
  • an amino acid with a polar side chain substituted for another amino acid with a polar side chain e.g., Asp or Glu
  • an amino acid with a nonpolar side chain substituted for another amino acid with a nonpolar side chain e.g., Ala, Gly, Val, Ile, Leu, Met, Phe, Pro, Trp, Val, etc
  • the functional variants can comprise the amino acid sequence of the parent TCR, polypeptide, or protein with at least one non-conservative amino acid substitution.
  • the non-conservative amino acid substitution it is preferable for the non-conservative amino acid substitution to not interfere with or inhibit the biological activity of the functional variant.
  • the non-conservative amino acid substitution enhances the biological activity of the functional variant, such that the biological activity of the functional variant is increased as compared to the parent TCR, polypeptide, or protein.
  • the TCR, polypeptide, or protein can consist essentially of the specified amino acid sequence or sequences described herein, such that other components of the TCR, Leydig 770310 HHS E-054-2023-0-PC-01 15 polypeptide, or protein, e.g., other amino do not materially change the biological activity of the TCR, polypeptide, or protein.
  • a polypeptide comprising a functional portion of any of the TCRs described herein.
  • the functional portion can be any portion comprising contiguous amino acids of the TCR of which it is a part, provided that the functional portion specifically binds to mutated p53.
  • Functional portions encompass, for example, those parts of a TCR that retain the ability to specifically bind to mutated p53 (e.g., in an applicable HLA molecule-dependent manner), or detect, treat, or prevent cancer, to a similar extent, the same extent, or to a higher extent, as the parent TCR.
  • the functional portion can comprise, for instance, about 10%, about 25%, about 30%, about 50%, about 70%, about 80%, about 90%, about 95%, or more, of the parent TCR.
  • the functional portion can comprise additional amino acids at the amino or carboxy terminus of the portion, or at both termini, which additional amino acids are not found in the amino acid sequence of the parent TCR.
  • the additional amino acids do not interfere with the biological function of the functional portion, e.g., specifically binding to mutated p53; and/or having the ability to detect cancer, treat or prevent cancer, etc. More desirably, the additional amino acids enhance the biological activity, as compared to the biological activity of the parent TCR.
  • the polypeptide can comprise a functional portion of either or both of the ⁇ and ⁇ chains of the TCRs of the invention, such as a functional portion comprising one of more of CDR1, CDR2, and CDR3 of the variable region(s) of the ⁇ chain and/or ⁇ chain of a TCR of the invention.
  • the polypeptide comprises the amino acid sequences of: (1) all of SEQ ID NOs: 2-7; (2) all of SEQ ID NOs: 16-21; (3) all of SEQ ID NOs: 30-35; (4) all of SEQ ID NOs: 44-49; (5) all of SEQ ID NOs: 58-63; (6) all of SEQ ID NOs: 72-77; or (7) all of SEQ ID NOs: 86-91.
  • the polypeptide may comprise the amino acid sequences of any one or more of: SEQ ID NOs: 2-7, 16-21, 30-35, 44-49, 58-63, 72-77, and 86-91.
  • polypeptide can comprise, for instance, the variable region of the inventive TCR comprising a combination of the CDR regions set forth above.
  • the polypeptide can comprise, e.g., the amino acid sequences of: (1) both of SEQ ID NOs: 8 and 9; (2) both of SEQ ID NOs: 10 and 11; (3) both of SEQ ID NOs: 22 and 23; (4) both of SEQ ID NOs: 24 and 25; (5) both of SEQ ID NOs: 36 and 37; (6) both of SEQ ID NOs: 38 and 39; (7) both of SEQ ID NOs: 50 and 51; (8) both of SEQ ID NOs: 52 and 53; (9) both of SEQ ID NOs: 64 and 65; (10) both of SEQ ID NOs: 66 and 67; (11) both of SEQ ID NOs: 78 and 79; (12) both of SEQ ID NOs: 80 and 81; (13) both of
  • the polypeptide may, e.g., comprise the amino acid sequence of any one or more of SEQ ID NOs: 8, 9, 10, 11, 22, 23, 24, 25, 36, 37, 38, 39, 50, 51, 52, 53, 64, 65, 66, 67, 78, 79, 80, 81, 92, 93, 94 and 95.
  • the polypeptide comprises the amino acid sequence(s) of: (1) SEQ ID NO: 8; (2) SEQ ID NO: 9; (3) both of SEQ ID NOs: 8 and 9; (4) SEQ ID NO: 10; (5) SEQ ID NO: 11; (6) both of SEQ ID NOs: 10 and 11; (7) SEQ ID NO: 22; (8) SEQ ID NO: 23; (9) both of SEQ ID NOs: 22 and 23; (10) SEQ ID NO: 24; (11) SEQ ID NO: 25; (12) both of SEQ ID NOs: 24 and 25; (13) SEQ ID NO: 36; (14) SEQ ID NOs: 37; (15) both of SEQ ID NOs: 36 and 37; (16) SEQ ID NO: 38; (17) SEQ ID NO: 39; (18) both of SEQ ID NOs: 38 and 39; (19) SEQ ID NO: 50; (20) SEQ ID NO: 51; (21) both of SEQ ID NOs: 50 and 51; (22) SEQ ID NO: 52; (23) SEQ ID NO: 10;
  • the inventive polypeptide can further comprise the constant region of the inventive TCR set forth above.
  • the polypeptide can comprise, e.g., the amino acid sequence of (i) one of SEQ ID NOs 100-102 or (ii) SEQ ID NO: 102 and one of SEQ ID NOs: 100 and 101.
  • the inventive polypeptide may comprise an ⁇ chain and a ⁇ chain of the inventive TCR.
  • the polypeptide can comprise, e.g., the amino acid sequences of: (1) both of SEQ ID NOs: 12 and 13; (2) both of SEQ ID NOs: 14 and 15; (3) both of SEQ ID NOs: 26 and 27; (4) both of SEQ ID NOs: 28 and 29; (5) both of Leydig 770310 HHS E-054-2023-0-PC-01 17 SEQ ID NOs: 40 and 41; (6) both of SEQ 42 and 43; (7) both of SEQ ID NOs: 54 and 55; (8) both of SEQ ID NOs: 56 and 57; (9) both of SEQ ID NOs: 68 and 69; (10) both of SEQ ID NOs: 70 and 71; (11) SEQ ID NO: 82 and 83; (12) SEQ ID NO: 84 and 85; (13) SEQ ID NO: 96 and 97; or (14) SEQ ID NO: 98 and 99.
  • the polypeptide may comprise the amino acid sequence of any one or more of SEQ ID NOs: 12, 13, 14, 15, 26, 27, 28, 29, 40, 41, 42, 43, 54, 55, 56, 57, 68, 69, 70, 71, 82, 83, 84, 85, 96, 97, 98, and 99.
  • the polypeptide comprises the amino acid sequences of: (1) SEQ ID NO: 12; (2) SEQ ID NO: 13; (3) both of SEQ ID NOs: 12 and 13; (4) SEQ ID NO: 14; (5) SEQ ID NO: 15; (6) both of SEQ ID NOs: 14 and 15; (7) SEQ ID NO: 26; (8) SEQ ID NO: 27; (9) both of SEQ ID NOs: 26 and 27; (10) SEQ ID NO: 28; (11) SEQ ID NO: 29; (12) both of SEQ ID NOs: 28 and 29; (13) SEQ ID NO: 40; (14) SEQ ID NO: 41; (15) both of SEQ ID NOs: 40 and 41; (16) SEQ ID NO: 42; (17) SEQ ID NO: 43; (18) both of SEQ ID NOs: 42 and 43; (19) SEQ ID NO: 54; (20) SEQ ID NO: 55; (21) both of SEQ ID NOs: 54 and 55; (22) SEQ ID NO: 56; (23) SEQ ID NO:
  • an aspect of the invention further provides a protein comprising a functional portion of any of the inventive TCRs described herein.
  • protein is meant a molecule comprising one or more polypeptide chains.
  • the protein of the invention can comprise: first and second polypeptide chains, wherein: (1) the first polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 2-4; (2) the second polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 5-7; (3) the first polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 2-4 and the second polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 5-7; (4) the first polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 16-18; (5) the second polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 19-21; (6) the first polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 16-18 and the second polypeptide chain comprises
  • the protein comprises first and second polypeptide chains, wherein: (1) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 8; (2) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 9; (3) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 8 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 9; (4) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 10; (5) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 11; (6) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 10 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 11; (7) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 22; (8) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 23; (9) the first Leydig 770310 HHS E-054-2023-0-PC-01 19 polypeptide chain comprises the amino acid
  • the protein comprises first and second polypeptide chains, wherein: (1) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 12; (2) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 13; (3) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 12 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 13; (4) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 14; (5) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 15; (6) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 14 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 15; (7) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 26; (8) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 27; (9) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 26 and the second polypeptide chain comprises the amino acid
  • the protein of the invention may be a TCR.
  • the first and/or second polypeptide chain(s) of the protein further comprise(s) other amino acid sequences, e.g., an amino acid sequence encoding an immunoglobulin or a portion thereof, then the inventive protein can be a fusion protein.
  • an aspect of the invention also provides a fusion protein comprising at least one of the inventive polypeptides described herein along with at least one other polypeptide.
  • the other polypeptide can exist as a separate polypeptide of the fusion protein, or can exist as a polypeptide, which is expressed in frame (in tandem) with one of the inventive polypeptides described herein.
  • the other polypeptide can encode any peptidic or proteinaceous molecule, or a portion thereof, including, but not limited to an immunoglobulin, CD3, CD4, CD8, an MHC molecule, a CD1 molecule, e.g., CD1a, CD1b, CD1c, CD1d, etc.
  • the fusion protein can comprise one or more copies of the inventive polypeptide and/or one or more copies of the other polypeptide.
  • the fusion protein can comprise 1, 2, 3, 4, 5, or more, copies of the inventive polypeptide and/or of the other polypeptide. Suitable methods of making fusion proteins are known in the art, and include, for example, recombinant methods.
  • the TCRs, polypeptides, and proteins of the invention may be expressed as a single protein comprising a linker peptide linking the ⁇ chain and the ⁇ chain.
  • the TCRs, polypeptides, and proteins of the invention may further comprise a linker peptide.
  • the linker peptide may advantageously facilitate the expression of a recombinant TCR, polypeptide, and/or protein in a host cell.
  • the linker peptide may comprise any suitable amino acid sequence.
  • the linker peptide may comprise the amino acid sequence of RAKRSGSGATNFSLLKQAGDVEENPGP (SEQ ID NO: 103).
  • the linker peptide may be cleaved, resulting in separated ⁇ and ⁇ chains.
  • the TCR, polypeptide, or protein may comprise an amino acid sequence comprising a full-length ⁇ chain, a full-length ⁇ chain, and a linker peptide positioned between the ⁇ and ⁇ chains.
  • Leydig 770310 HHS E-054-2023-0-PC-01 23 the TCR, or protein disclosed herein comprises an ⁇ chain and/or a ⁇ chain, as disclosed herein, comprising a signal peptide.
  • the sequence of the signal peptide of any of the ⁇ chains and/or ⁇ chains disclosed herein comprises an leucine, lysine, alanine or histidine residue substituted for the wild-type residue at position 2.
  • the TCR, polypeptide or protein disclosed herein comprises a mature version of an ⁇ chain and/or a ⁇ chain, as disclosed herein, that lacks a signal peptide.
  • the protein of the invention can be a recombinant antibody, or an antigen binding portion thereof, comprising at least one of the inventive polypeptides described herein.
  • recombinant antibody refers to a recombinant (e.g., genetically engineered) protein comprising at least one of the polypeptides of the invention and a polypeptide chain of an antibody, or an antigen binding portion thereof.
  • the polypeptide of an antibody, or antigen binding portion thereof can be a heavy chain, a light chain, a variable or constant region of a heavy or light chain, a single chain variable fragment (scFv), or an Fc, Fab, or F(ab)2' fragment of an antibody, etc.
  • the polypeptide chain of an antibody, or an antigen binding portion thereof can exist as a separate polypeptide of the recombinant antibody.
  • the polypeptide chain of an antibody, or an antigen binding portion thereof can exist as a polypeptide, which is expressed in frame (in tandem) with the polypeptide of the invention.
  • the polypeptide of an antibody, or an antigen binding portion thereof can be a polypeptide of any antibody or any antibody fragment, including any of the antibodies and antibody fragments described herein.
  • the TCRs, polypeptides, and proteins of the invention can be of any length, i.e., can comprise any number of amino acids, provided that the TCRs, polypeptides, or proteins retain their biological activity, e.g., the ability to specifically bind to mutated p53; detect cancer in a mammal; or treat or prevent cancer in a mammal, etc.
  • the polypeptide can be in the range of from about 50 to about 5000 amino acids long, such as 50, 70, 75, 100, 125, 150, 175, 200, 300, 400, 500, 600, 700, 800, 900, 1000 or more amino acids in length.
  • the polypeptides of the invention also include oligopeptides.
  • the TCRs, polypeptides, and proteins of the invention of the invention can comprise synthetic amino acids in place of one or more naturally-occurring amino acids.
  • Such synthetic amino acids include, for example, aminocyclohexane carboxylic acid, norleucine, ⁇ -amino n-decanoic acid, homoserine, S-acetylaminomethyl- cysteine, trans-3- and trans-4-hydroxyproline, 4-aminophenylalanine, 4-nitrophenylalanine, Leydig 770310 HHS E-054-2023-0-PC-01 24 4-chlorophenylalanine, 4- ⁇ -phenylserine ⁇ -hydroxyphenylalanine, phenylglycine, ⁇ -naphthylalanine, cyclohexylalanine, cyclohexylglycine, indoline-2- carboxylic acid, 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid, aminomalonic acid, aminomalonic acid monoamide, N’-benzyl-N’-methyl-lysine, N’,N’-d
  • the TCRs, polypeptides, and proteins of the invention can be, e.g., glycosylated, amidated, carboxylated, phosphorylated, esterified, N-acylated, cyclized via, e.g., a disulfide bridge, or converted into an acid addition salt and/or optionally dimerized or polymerized, or conjugated.
  • inventive TCRs, polypeptides, and proteins described herein are also contemplated to be useful as the soluble TCR component of bispecific engager TCR fusion proteins (e.g., IMMTAC (immune- mobilizing monoclonal TCRs against cancer) molecules).
  • Bispecific engager TCR fusion proteins have two components.
  • the other component comprises an anti-CD3 effector.
  • the anti-CD3 effector may be any molecule that engages with a CD3 molecule on T cells and activates a T cell immune response.
  • the anti-CD3 effector may be an anti-CD3 antibody or anti-CD3 antibody fragment.
  • the soluble TCR component of the bispecific engager TCR fusion protein binds to the target antigen presented on the surface of cancer cells presented by an HLA molecule.
  • the anti- CD3 effector component engages a CD3 molecule on T cells. The engagement of these components of the bispecific engager TCR fusion protein triggers the activation and recruitment of T cells and redirects T-cell killing to tumor cells.
  • An aspect of the invention provides a bispecific engager TCR fusion protein comprising (i) any of the inventive TCRs, polypeptides, or proteins (including any of the functional portions or variants thereof) described herein and (ii) an anti-CD3 effector.
  • references to “protein(s)” also encompass the bispecific engager TCR fusion proteins described herein, unless specified otherwise.
  • the TCR, polypeptide, and/or protein of the invention can be obtained by methods known in the art such as, for example, de novo synthesis.
  • polypeptides and proteins can be recombinantly produced using the nucleic acids described herein using standard Leydig 770310 HHS E-054-2023-0-PC-01 25 recombinant methods.
  • TCRs, polypeptides, and/or proteins described herein can be synthesized by any of a variety of commercial entities.
  • inventive TCRs, polypeptides, and proteins can be synthetic, recombinant, isolated, and/or purified.
  • An aspect of the invention provides a nucleic acid comprising a nucleotide sequence encoding any of the TCRs, polypeptides, or proteins described herein.
  • Nucleic acid includes “polynucleotide,” “oligonucleotide,” and “nucleic acid molecule,” and generally means a polymer of DNA or RNA, which can be single-stranded or double-stranded, which can contain natural, non-natural or altered nucleotides, and which can contain a natural, non-natural or altered internucleotide linkage, such as a phosphoroamidate linkage or a phosphorothioate linkage, instead of the phosphodiester found between the nucleotides of an unmodified oligonucleotide.
  • the nucleic acid comprises complementary DNA (cDNA).
  • the nucleic acid does not comprise any insertions, deletions, inversions, and/or substitutions. However, it may be suitable in some instances, as discussed herein, for the nucleic acid to comprise one or more insertions, deletions, inversions, and/or substitutions.
  • An aspect of the invention provides an isolated or purified nucleic acid comprising, from 5’ to 3’, a first nucleic acid sequence and a second nucleotide sequence, wherein the first and second nucleotide sequence, respectively, encode the amino sequences of SEQ ID NOs: 8 and 9; 9 and 8; 10 and 11; 11 and 10; 12 and 13; 13 and 12; 14 and 15; 15 and 14; 22 and 23; 23 and 22; 24 and 25; 25 and 24; 26 and 27; 27 and 26; 28 and 29; 29 and 28; 36 and 37; 37 and 36; 38 and 39; 39 and 38; 40 and 41; 41 and 40; 42 and 43; 43 and 42; 50 and 51; 51 and 50; 52 and 53; 53 and 52; 54 and 55; 55 and 54; 56 and 57; 57 and 56; 64 and 65; 65 and 64; 66 and 67; 67 and 66; 68 and 69; 69 and 68; 70 and 71; 71 and 70; 78 and 79; 79 and 78
  • the nucleic acid further comprises a third nucleotide acid sequence interposed between the first and second nucleotide sequence, wherein the third nucleotide sequence encodes a cleavable linker peptide.
  • the cleavable linker peptide may comprise the amino acid sequence of RAKRSGSGATNFSLLKQAGDVEENPGP (SEQ ID NO: 103).
  • the nucleic acids of invention are recombinant.
  • the term “recombinant” refers to (i) molecules that are constructed outside living cells by joining natural or synthetic nucleic acid segments to nucleic acid molecules that can replicate in a living cell, or (ii) molecules that result from the replication of those described in (i) above.
  • the replication can be in vitro replication or in vivo replication.
  • the nucleic acids can be constructed based on chemical synthesis and/or enzymatic ligation reactions using procedures known in the art. See, for example, Green and Sambrook et al., supra.
  • a nucleic acid can be chemically synthesized using naturally occurring nucleotides or variously modified nucleotides designed to increase the biological stability of the molecules or to increase the physical stability of the duplex formed upon hybridization (e.g., phosphorothioate derivatives and acridine substituted nucleotides).
  • modified nucleotides that can be used to generate the nucleic acids include, but are not limited to, 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xanthine, 4-acetylcytosine, 5-(carboxyhydroxymethyl) uracil, 5-carboxymethylaminomethyl- 2-thiouridine, 5-carboxymethylaminomethyluracil, dihydrouracil, beta-D-galactosylqueosine, inosine, N 6 -isopentenyladenine, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2- methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, N 6 -substituted adenine, 7-methylguanine, 5-methylaminomethyluracil, 5-methoxyaminomethyl-2-thiouracil, beta-D-mannosyl
  • nucleic acid comprising a nucleotide sequence which is complementary to the nucleotide sequence of any of the nucleic acids described herein.
  • the nucleic acids of the invention can be incorporated into a recombinant expression vector.
  • an aspect of the invention provides a recombinant expression vector comprising any of the nucleic acids of the invention.
  • the recombinant expression vector comprises a nucleotide sequence encoding the ⁇ chain, the ⁇ chain, and linker peptide.
  • the term “recombinant expression vector” means a genetically-modified oligonucleotide or polynucleotide construct that permits the expression of an mRNA, protein, polypeptide, or peptide by a host cell, when the construct comprises a nucleotide sequence encoding the mRNA, protein, polypeptide, or peptide, and the vector is contacted with the cell under conditions sufficient to have the mRNA, protein, polypeptide, or peptide expressed within the cell.
  • the vectors of the invention are not naturally-occurring as a whole. However, parts of the vectors can be naturally-occurring.
  • the inventive recombinant expression vectors can comprise any type of nucleotide, including, but not limited to DNA and RNA, which can be single-stranded or double-stranded, synthesized or obtained in part from natural sources, and which can contain natural, non-natural or altered nucleotides.
  • the recombinant expression vectors can comprise naturally-occurring, non- naturally-occurring internucleotide linkages, or both types of linkages.
  • the non- naturally occurring or altered nucleotides or internucleotide linkages do not hinder the transcription or replication of the vector.
  • the recombinant expression vector of the invention can be any suitable recombinant expression vector, and can be used to transform or transfect any suitable host cell.
  • Suitable vectors include those designed for propagation and expansion or for expression or both, such as plasmids and viruses.
  • the vector can be selected from the group consisting of the transposon/transposase series, pUC series (Fermentas Life Sciences), the pBluescript series (Stratagene, LaJolla, CA), the pET series (Novagen, Madison, WI), the pGEX series (Pharmacia Biotech, Uppsala, Sweden), and the pEX series (Clontech, Palo Alto, CA).
  • Bacteriophage vectors such as ⁇ GT10, ⁇ GT11, ⁇ ZapII (Stratagene), ⁇ EMBL4, and ⁇ NM1149, also can be used.
  • animal expression vectors include pEUK-Cl, pMAM and pMAMneo (Clontech).
  • the recombinant expression vector is a transposon or a viral vector, e.g., a lentiviral vector or a retroviral vector.
  • Leydig 770310 HHS E-054-2023-0-PC-01 28 The recombinant expression of the invention can be prepared using standard recombinant DNA techniques described in, for example, Green and Sambrook et al., supra. Constructs of expression vectors, which are circular or linear, can be prepared to contain a replication system functional in a prokaryotic or eukaryotic host cell.
  • Replication systems can be derived, e.g., from ColEl, 2 ⁇ plasmid, ⁇ , SV40, bovine papillomavirus, and the like.
  • the recombinant expression vector comprises regulatory sequences, such as transcription and translation initiation and termination codons, which are specific to the type of host cell (e.g., bacterium, fungus, plant, or animal) into which the vector is to be introduced, as appropriate and taking into consideration whether the vector is DNA- or RNA- based.
  • the recombinant expression vector can include one or more marker genes, which allow for selection of transformed or transfected host cells.
  • Marker genes include biocide resistance, e.g., resistance to antibiotics, heavy metals, etc., complementation in an auxotrophic host cell to provide prototrophy, and the like.
  • Suitable marker genes for the inventive expression vectors include, for instance, neomycin/G418 resistance genes, hygromycin resistance genes, histidinol resistance genes, tetracycline resistance genes, and ampicillin resistance genes.
  • the recombinant expression vector can comprise a native or nonnative promoter operably linked to the nucleotide sequence encoding the TCR, polypeptide, or protein, or to the nucleotide sequence which is complementary to the nucleotide sequence encoding the TCR, polypeptide, or protein.
  • the selection of promoters is within the ordinary skill of the artisan.
  • the combining of a nucleotide sequence with a promoter is also within the skill of the artisan.
  • the promoter can be a non-viral promoter, e.g., a human elongation factor-1 ⁇ promoter, or a viral promoter, e.g., a cytomegalovirus (CMV) promoter, an SV40 promoter, an RSV promoter, and a promoter found in the long-terminal repeat of the murine stem cell virus.
  • CMV cytomegalovirus
  • the inventive recombinant expression vectors can be designed for either transient expression, for stable expression, or for both.
  • the recombinant expression vectors can be made for constitutive expression or for inducible expression. Further, the recombinant expression vectors can be made to include a suicide gene.
  • suicide gene refers to a gene that causes the cell expressing Leydig 770310 HHS E-054-2023-0-PC-01 29 the suicide gene to die.
  • the suicide gene can a gene that confers sensitivity to an agent, e.g., a drug, upon the cell in which the gene is expressed, and causes the cell to die when the cell is contacted with or exposed to the agent.
  • host cell refers to any type of cell that can contain the inventive recombinant expression vector.
  • the host cell can be a eukaryotic cell, e.g., plant, animal, fungi, or algae, or can be a prokaryotic cell, e.g., bacteria or protozoa.
  • the host cell can be a cultured cell or a primary cell, i.e., isolated directly from an organism, e.g., a human.
  • the host cell can be an adherent cell or a suspended cell, i.e., a cell that grows in suspension.
  • Suitable host cells are known in the art and include, for instance, DH5 ⁇ E. coli cells, Chinese hamster ovarian cells, monkey VERO cells, COS cells, HEK293 cells, and the like.
  • the host cell is preferably a prokaryotic cell, e.g., a DH5 ⁇ cell.
  • the host cell is preferably a mammalian cell. Most preferably, the host cell is a human cell.
  • the host cell may be a human lymphocyte.
  • the host cell is selected from the group consisting of a T cell, a natural killer T (NKT) cell, an invariant natural killer T (iNKT) cell, a natural killer (NK) cell, a macrophage, a pluripotent cell, and a multipotent cell.
  • the host cell can be of any cell type, can originate from any type of tissue, and can be of any developmental stage, the host cell preferably is a peripheral blood lymphocyte (PBL) or a peripheral blood mononuclear cell (PBMC). More preferably, the host cell is a T cell.
  • PBL peripheral blood lymphocyte
  • PBMC peripheral blood mononuclear cell
  • the T cell can be any T cell, such as a cultured T cell, e.g., a primary T cell, or a T cell from a cultured T cell line, e.g., Jurkat, SupT1, etc., or a T cell obtained from a mammal. If obtained from a mammal, the T cell can be obtained from Leydig 770310 HHS E-054-2023-0-PC-01 30 numerous sources, including but not limited blood, bone marrow, lymph node, the thymus, or other tissues or fluids. T cells can also be enriched for or purified. Preferably, the T cell is a human T cell.
  • Pluripotent cells may comprise, for example, stem cells, e.g., embryonic stem cells, nuclear transfer derived embryonic stem cells, induced pluripotent stem cells (iPSC), etc.
  • Multipotent cells may comprise, for example, hematopoietic stem cells.
  • Modifying, e.g., reprogramming, cells to a pluripotent state refers to the reversion of a cell to a pluripotent cell and is described for example, in Crompton et al., Trends Immunol., 35(4): 178-185 (2014).
  • Exemplary techniques may include somatic cell nuclear transfer (SCNT), cell–cell fusion, and direct reprogramming.
  • Examples of methods for carrying out cell-cell fusion are described, for example, in Ogle et al., Nat. Rev. Mol. Cell Biol.6: 567-75 (2005) and Zhou et al., Cell Stem Cell, 3: 382-388 (2008).
  • Examples of methods for carrying out SCNT are described, for example, in Hanna et al., Cell, 143: 508- 525 (2010); Stadtfeld et al., Genes Dev., 24: 2239-2263 (2010); Wilmut et al., Nature, 385: 810-813 (1997); Vizcardo et al., Cell Stem Cell, 12: 31-36 (2013); and Crompton et al., Cell Stem Cell, 12: 6-8 (2013).
  • the host cell is an iPSC that was prepared by reprogramming, any of the host cells described herein (e.g., T cells, NK cells, or invariant natural killer T cells) to a pluripotent state.
  • a population of cells comprising at least one host cell described herein.
  • the population of cells can be a heterogeneous population comprising the host cell comprising any of the recombinant expression vectors described, in addition to at least one other cell, e.g., a host cell (e.g., a T cell), which does not comprise any of the recombinant expression vectors, or a cell other than a T cell, e.g., a B cell, a macrophage, a neutrophil, an erythrocyte, a hepatocyte, an endothelial cell, an epithelial cell, a muscle cell, a brain cell, etc.
  • a host cell e.g., a T cell
  • a cell other than a T cell e.g., a B cell, a macrophage, a neutrophil, an erythrocyte, a hepatocyte, an endothelial cell, an epithelial cell, a muscle cell, a brain cell, etc.
  • Expansion of the numbers of T cells can be accomplished by any of a number of methods as are known in the art as described in, for example, U.S. Patent Nos. 8,034,334; 8,383,099; and 11,401,503; Dudley et al., J. Immunother., 26:332-42 (2003); and Riddell et al., J. Immunol. Methods, 128:189-201 (1990).
  • expansion of the numbers of T cells is carried out by culturing the T cells with OKT3 antibody, IL-2, and feeder PBMC (e.g., irradiated allogeneic PBMC).
  • Another aspect of the invention provides a method of producing an engineered human cell (or an engineered population of human cells), the method comprising introducing any of the inventive nucleic acids or recombinant expression vectors described herein to an isolated human cell (or an isolated population of human cells), wherein the nucleic acid or recombinant expression vector comprises a nucleotide sequence encoding any of the inventive TCRs, polypeptides, or proteins described herein.
  • the isolated human cell, or isolated population of human cells, to which the nucleic acid or recombinant expression vector is introduced may be as described herein with respect to other aspects of the invention.
  • the inventive TCRs, polypeptides, proteins, nucleic acids, recombinant expression vectors, and host cells can be isolated and/or purified.
  • isolated as used herein means having been removed from its natural Leydig 770310 HHS E-054-2023-0-PC-01 32 environment.
  • purified as used means having been increased in purity, wherein “purity” is a relative term, and not to be necessarily construed as absolute purity.
  • the purity can be at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or can be about 100%.
  • inventive TCRs, polypeptides, proteins, nucleic acids, recombinant expression vectors, and host cells (including populations thereof), all of which are collectively referred to as “inventive TCR materials” hereinafter, can be formulated into a composition, such as a pharmaceutical composition.
  • a pharmaceutical composition comprising any of the TCRs, polypeptides, proteins, nucleic acids, expression vectors, and host cells (including populations thereof), described herein, and a pharmaceutically acceptable carrier.
  • inventive pharmaceutical compositions containing any of the inventive TCR materials can comprise more than one inventive TCR material, e.g., a polypeptide and a nucleic acid, or two or more different TCRs.
  • the pharmaceutical composition can comprise an inventive TCR material in combination with another pharmaceutically active agent(s) or drug(s), such as a chemotherapeutic agent, e.g., asparaginase, busulfan, carboplatin, cisplatin, daunorubicin, doxorubicin, fluorouracil, gemcitabine, hydroxyurea, methotrexate, paclitaxel, rituximab, vinblastine, vincristine, etc.
  • a chemotherapeutic agent e.g., asparaginase, busulfan, carboplatin, cisplatin, daunorubicin, doxorubicin, fluorouracil, gemcitabine, hydroxyurea, methotrexate, paclitaxel, rituximab, vinblastine, vincristine, etc.
  • the carrier is a pharmaceutically acceptable carrier.
  • the carrier can be any of those conventionally used for the particular inventive TCR material under consideration
  • compositions of the invention there are a variety of suitable formulations of the pharmaceutical composition of the invention. Suitable formulations may include any of those for parenteral, subcutaneous, intravenous, intramuscular, intraarterial, intrathecal, intratumoral, or interperitoneal administration.
  • More than one route can be used to administer the inventive TCR materials, and in certain instances, a particular route can provide a more immediate and more effective response than another route.
  • Leydig 770310 HHS E-054-2023-0-PC-01 33 Preferably, the inventive TCR is administered by injection, e.g., intravenously.
  • the pharmaceutically acceptable carrier for the cells for injection may include any isotonic carrier such as, for example, normal saline (about 0.90% w/v of NaCl in water, about 300 mOsm/L NaCl in water, or about 9.0 g NaCl per liter of water), NORMOSOL R electrolyte solution (Abbott, Chicago, IL), PLASMA-LYTE A (Baxter, Deerfield, IL), about 5% dextrose in water, or Ringer's lactate.
  • the pharmaceutically acceptable carrier is supplemented with human serum albumin.
  • the amount or dose (e.g., numbers of cells when the inventive TCR material is one or more cells) of the inventive TCR material administered should be sufficient to effect, e.g., a therapeutic or prophylactic response, in the subject or animal over a reasonable time frame.
  • the dose of the inventive TCR material should be sufficient to bind to a cancer antigen (e.g., mutated p53), or detect, treat or prevent cancer in a period of from about 2 hours or longer, e.g., 12 to 24 or more hours, from the time of administration. In certain aspects, the time period could be even longer.
  • the dose will be determined by the efficacy of the particular inventive TCR material and the condition of the animal (e.g., human), as well as the body weight of the animal (e.g., human) to be treated.
  • Many assays for determining an administered dose are known in the art. For example, an assay, which comprises comparing the extent to which target cells are lysed or IFN- ⁇ is secreted by T cells expressing the inventive TCR, polypeptide, or protein upon administration of a given dose of such T cells to a mammal among a set of mammals of which each is given a different dose of the T cells, could be used to determine a starting dose to be administered to a mammal.
  • inventive TCR material The extent to which target cells are lysed or IFN- ⁇ is secreted upon administration of a certain dose can be assayed by methods known in the art.
  • the dose of the inventive TCR material also will be determined by the existence, nature and extent of any adverse side effects that might accompany the administration of a particular inventive TCR material.
  • the attending physician will decide the dosage of the inventive TCR material with which to treat each individual patient, taking into consideration a variety of factors, such as age, body weight, general health, diet, sex, inventive TCR material to be administered, route of administration, and the severity of the cancer being treated.
  • the number of cells administered per infusion may vary, e.g., from about 1 x 10 6 to about 1 x 10 12 cells or more. In certain aspects, fewer than 1 x 10 6 cells may be administered.
  • Leydig 770310 HHS E-054-2023-0-PC-01 34 One of ordinary skill in the art readily appreciate that the inventive TCR materials of the invention can be modified in any number of ways, such that the therapeutic or prophylactic efficacy of the inventive TCR materials is increased through the modification.
  • the inventive TCR materials can be conjugated either directly or indirectly through a bridge to a chemotherapeutic agent. The practice of conjugating compounds to a chemotherapeutic agent is known in the art.
  • inventive TCR materials which are not necessary for the function of the inventive TCR materials, are ideal sites for attaching a bridge and/or a chemotherapeutic agent, provided that the bridge and/or chemotherapeutic agent, once attached to the inventive TCR materials, do(es) not interfere with the function of the inventive TCR materials, i.e., the ability to bind to mutated p53 or to detect, treat, or prevent cancer.
  • inventive pharmaceutical compositions, TCRs, polypeptides, proteins, nucleic acids, recombinant expression vectors, host cells, or populations of cells can be used in methods of treating or preventing cancer.
  • inventive TCRs are believed to bind specifically to mutated p53, such that the TCR (or related inventive polypeptide or protein), when expressed by a cell, is able to mediate an immune response against a target cell expressing mutated p53.
  • an aspect of the invention provides a method of treating or preventing cancer in a mammal, comprising administering to the mammal any of the pharmaceutical compositions, TCRs, polypeptides, or proteins described herein, any nucleic acid or recombinant expression vector comprising a nucleotide sequence encoding any of the TCRs, polypeptides, proteins described herein, or any host cell or population of cells comprising a recombinant vector which encodes any of the TCRs, polypeptides, or proteins described herein, in an amount effective to treat or prevent cancer in the mammal.
  • An aspect of the invention provides any of the pharmaceutical compositions, TCRs, polypeptides, or proteins described herein, any nucleic acid or recombinant expression vector comprising a nucleotide sequence encoding any of the TCRs, polypeptides, proteins described herein, or any host cell or population of cells comprising a recombinant vector which encodes any of the TCRs, polypeptides, or proteins described herein, for use in the treatment or prevention of cancer in a mammal.
  • the terms “treat,” and “prevent” as well as words stemming therefrom, as used herein, do not necessarily imply 100% or complete treatment or prevention.
  • the inventive methods can provide any amount of any level of treatment or prevention of cancer in a mammal.
  • the treatment or prevention provided by the inventive method can include treatment or prevention of one or more conditions or symptoms of the cancer being treated or prevented.
  • treatment or prevention can include promoting the regression of a tumor.
  • prevention can encompass delaying the onset of the cancer, or a symptom or condition thereof. Alternatively or additionally, “prevention” may encompass preventing or delaying the recurrence of cancer, or a symptom or condition thereof.
  • inventive pharmaceutical compositions TCRs, polypeptides, proteins, nucleic acids, recombinant expression vectors, host cells, or populations of cells can be used in methods of inducing an immune response against a cancer in a mammal.
  • an aspect of the invention provides a method of inducing an immune response against a cancer in a mammal, comprising administering to the mammal any of the pharmaceutical compositions, TCRs, polypeptides, or proteins described herein, any nucleic acid or recombinant expression vector comprising a nucleotide sequence encoding any of the TCRs, polypeptides, proteins described herein, or any host cell or population of cells comprising a recombinant vector which encodes any of the TCRs, polypeptides, or proteins described herein, in an amount effective to induce an immune response against the cancer in the mammal.
  • An aspect of the invention provides any of the pharmaceutical compositions, TCRs, polypeptides, or proteins described herein, any nucleic acid or recombinant expression vector comprising a nucleotide sequence encoding any of the TCRs, polypeptides, proteins described herein, or any host cell or population of cells comprising a recombinant vector which encodes any of the TCRs, polypeptides, or proteins described herein, for use in the inducement of an immune response against a cancer in a mammal. Also provided by an aspect of the invention is a method of detecting the presence of cancer in a mammal.
  • the method comprises (i) contacting a sample comprising one or more cells from the mammal with any of the inventive TCRs, polypeptides, proteins, nucleic acids, recombinant expression vectors, host cells, populations of cells, or pharmaceutical compositions described herein, thereby forming a complex, and (ii) detecting the complex, wherein detection of the complex is indicative of the presence of cancer in the mammal.
  • the sample of cells can be a sample comprising whole cells, lysates thereof, or a fraction of the whole cell lysates, e.g., a nuclear or cytoplasmic fraction, a whole protein fraction, or a nucleic acid fraction.
  • the contacting can take place in vitro or in vivo with respect to the mammal.
  • the contacting is in vitro.
  • detection of the complex can occur through any number of ways known in the art.
  • the inventive TCRs, polypeptides, proteins, nucleic acids, recombinant expression vectors, host cells, or populations of cells, described herein can be labeled with a detectable label such as, for instance, a radioisotope, a fluorophore (e.g., fluorescein isothiocyanate (FITC), phycoerythrin (PE)), an enzyme (e.g., alkaline phosphatase, horseradish peroxidase), and element particles (e.g., gold particles).
  • a detectable label such as, for instance, a radioisotope, a fluorophore (e.g., fluorescein isothiocyanate (FITC), phycoerythrin (PE)), an enzyme (e.g., alkaline phosphatase, horseradish peroxidase), and element particles (e.g., gold particles).
  • a detectable label such as, for instance, a radioiso
  • the cells are autologous to the mammal.
  • the cancer can be any cancer, including, e.g., any of acute lymphocytic cancer, acute myeloid leukemia, alveolar rhabdomyosarcoma, bone cancer, brain cancer, breast cancer, cancer of the anus, anal canal, or anorectum, cancer of the eye, cancer of the intrahepatic bile duct, cancer of the joints, cancer of the neck, gallbladder, or pleura, cancer of the nose, nasal cavity, or middle ear, cancer of the oral cavity, cancer of the vagina, cancer of the vulva, chronic lymphocytic leukemia, chronic myeloid cancer, colon cancer, colocrectal cancer, endometrial cancer, esophageal cancer, uterine cervical cancer, gastrointestinal carcinoid tumor, glioma, Hodgkin lymphoma, hypopharynx cancer, kidney cancer, larynx cancer, liver cancer, lung cancer,
  • the cancer is a cancer which expresses mutated p53.
  • the cancer may express p53 with a mutation at one or both of positions 220 and 175, as defined by SEQ ID NO: 1.
  • the cancer may express p53 with one or both of the following human p53 mutations: Y220C and R175H.
  • the cancer is an epithelial cancer.
  • the cancer is colon cancer, rectal cancer, ovarian cancer, endometrial cancer, non-small cell lung cancer (NSCLC), glioblastoma, uterine cervical cancer, head and neck cancer, breast cancer, pancreatic cancer, or bladder cancer.
  • NSCLC non-small cell lung cancer
  • the cancer may be known to comprise a Y220C or R175H mutation in human p53.
  • the mammal referred to in the inventive methods can be any mammal.
  • the term “mammal” refers to any mammal, including, but not limited to, mammals of the order Rodentia, such as mice and hamsters, and mammals of the order Lagomorpha, such as rabbits. It is preferred that the mammals are from the order Carnivora, including Felines (cats) and Canines (dogs). It is more preferred that the mammals are from the order Artiodactyla, including Bovines (cows) and Swines (pigs) or of the order Perssodactyla, including Equines (horses).
  • EXAMPLE 1 This example demonstrates the isolation of TCRs from Patient 4196.
  • the p53 R175H 9-amino acid epitope (HMTEVVRHC) (SEQ ID NO: 112) contains cysteine at its C-terminus.
  • Cysteine residues can oxidize and form disulfide bonds with other cysteine residues under oxidizing conditions, thus potentially interfering with their ability to bind to MHC molecules (Sachs et al., J. Immunol., 205(2): 539–549 (2020)). This may lead to reduced T cell activity.
  • HMTEVVRHC HMTEVVRHC
  • AABA alpha-aminobutyric acid
  • AABA is a cysteine analogue containing a methyl group in place of the sulfhydryl group present in cysteine.
  • the native cysteine at position 9 of (HMTEVVRRC) (SEQ ID NO: 113) was replaced with AABA to produce the corresponding wild-type analogue peptide HMTEVVRRX (SEQ ID NO: 117), wherein X at position 9 of SEQ ID NO: 117 is AABA.
  • Three different culture conditions were tested prior to isolating TCRs specific for p53 R175H from TILs from Patient 4196 (4196 TILs).
  • the HLA-A2 tetramer was oligomerized HLA-A2 monomers loaded with the p53 R175H 9-amino acid epitope analogue HMTEVVRHX (SEQ ID NO: 116), wherein X at position 9 of SEQ ID NO: 116 is AABA.
  • SEQ ID NO: 116 9-amino acid epitope analogue HMTEVVRHX
  • TCR alpha and beta chain variable regions were identified by single-cell TCR sequencing.
  • the amino acid sequences of the alpha and beta chain variable regions are shown in Table 2.
  • the CDRs are underlined.
  • the N-terminal signal peptides are in bold font.
  • T cells were Leydig 770310 HHS E-054-2023-0-PC-01 40 stained for the HLA-A2 tetramer.
  • the tetramer was oligomerized HLA-A2 loaded with the p53 R175H 9-amino acid epitope analogue HMTEVVRHX (SEQ ID NO: 116), wherein X at position 9 of SEQ ID NO: 116 is AABA.
  • the flow cytometry staining for CD8 and tetramer is shown in Figure 2A.
  • the 4424-R175H TCR was isolated from the tetramer.
  • the 4424-R175H TCR was the first TCR that was identified by use of a tetramer.
  • the sequences of the TCR alpha and beta chain variable regions were identified by single-cell TCR sequencing. The amino acid sequences of the alpha and beta chain variable regions are shown in Table 3. The CDRs are underlined. The N-terminal signal peptides are in bold font.
  • a R S T This example demonstrates the isolation of TCRs from Patient 4402.
  • TIL from Patient 4402 showed specific recognition of antigen presenting cells (APCs) pulsed with p53 Y220C peptide (DRNTFRHSVVVPCEPPEVGSDCTTI) (SEQ ID NO: 114), but not the corresponding WT peptide (DRNTFRHSVVVPYEPPEVGSDCTTI) (SEQ Leydig 770310 HHS E-054-2023-0-PC-01 41 ID NO: 115).
  • APCs antigen presenting cells
  • DRNTFRHSVVVPCEPPEVGSDCTTI p53 Y220C peptide
  • DRNTFRHSVVVPYEPPEVGSDCTTI SEQ Leydig 770310 HHS E-054-2023-0-PC-01 41
  • TCRs Twenty-one (21) TCRs (4402-Y220C-A to -U, respectively) were isolated from TIL from Patient 4402. ELISpot analysis was performed to detect IFN- ⁇ release following co-culture of healthy donor PBL expressing the respective 21 TCRs that were isolated from Patient 4402 TIL (effector cells).
  • the 4402-Y220C-B TCR, 4402-Y220C-C TCR, and 4402-Y220C-L TCR showed increased IFN- ⁇ release against B cells pulsed with 1 ⁇ g/mL mutant p53 Y220C peptide (DRNTFRHSVVVPCEPPEVGSDCTTI) (SEQ ID NO: 114) (target cells) relative to the DMSO controls, while the other 18 TCRs did not. Effector cells treated with phorbol 12- myristate 13-acetate (PMA) and ionomycin were used as a positive control. Controls included: cultures with only effector cells, cultures with target cells pulsed with DMSO, and cultures with PBL transduced with an empty vector (mock).
  • PMA phorbol 12- myristate 13-acetate
  • the sequences of the TCR alpha and beta chain variable regions were identified by single-cell TCR sequencing.
  • the amino acid sequences of the alpha and beta chain variable regions of three of the TCRs (4402-Y220C-B TCR (TRAV12-3/TRBV7-2), 4402- Y220C-C TCR (TRAV26-2/TRBV5-6), and 4402-Y220C-L TCR (TRAV23/TRBV5-1)) are shown in Table 4.
  • the CDRs are underlined.
  • the N-terminal signal peptides are in bold font.
  • Nucleotide sequences encoding the variable regions of the ⁇ and ⁇ chains of the TCRs of Tables 2-4 were obtained and codon optimized.
  • the TCR ⁇ VDJ regions were fused to the mouse TCR ⁇ constant chain.
  • the TCR ⁇ VJ regions were fused to the mouse TCR ⁇ constant chain.
  • Leydig 770310 HHS E-054-2023-0-PC-01 43 replacing the constant regions of the human and TCR ⁇ chains with the corresponding murine constant regions improves TCR expression and functionality (Cohen et al., Cancer Res., 66(17): 8878-86 (2006)).
  • the murine TCR ⁇ and TCR ⁇ constant chains were cysteine-modified.
  • TCR ⁇ and TCR ⁇ chains were separated by a Furin Ser/Gly P2A linker RAKRSGSGATNFSLLKQAGDVEENPGP (SEQ ID NO: 103). Without being bound to a particular theory or mechanism, it is believed that the linker provides comparable expression efficiency of the two chains (Szymczak et al., Nat. Biotechnol., 22(5):589-94 (2004)).
  • EXAMPLE 5 This example demonstrates the specificity and avidity of the 4196-C TCR, 4196-D TCR, and 4196-E TCR.
  • Healthy donor PBL were transduced with the retroviral vector encoding the 4196- C TCR, 4196-D TCR, or 4196-E TCR of Example 4 or the TCR 4196_AV6_with_BV11-2 disclosed in US 2020/0277352.
  • TCR-expressing T cells were co-cultured for 18 hours (h) with T2 cells that expressed HLA-A2 and had been incubated with the WT epitope analogue HMTEVVRRX (SEQ ID NO: 117), wherein X at position 9 of SEQ ID NO: 117 is AABA, or p53 R175H 9-amino acid epitope analogue at the concentration of peptides denoted Leydig 770310 HHS E-054-2023-0-PC-01 48 in Figures 1B-1E.
  • T cell activation marker was measured by flow cytometry following the co-culture.
  • the 4196-C TCR, 4196-D TCR, and 4196-E TCR all showed specificity for the mutant p53 epitope analogue.
  • the TCR 4196_AV6_with_BV11-2 showed significant wild-type epitope analogue recognition at the 1000 ng/mL and 100 ng/mL concentration.
  • Effector cells were prepared by transducing healthy donor PBL with the retroviral vector encoding the 4196-C TCR, 4196-D TCR, or 4196-E TCR of Example 4.
  • the anti- tumor activity of the 4196-C TCR, 4196-D TCR, and 4196-E TCR was determined using an INCUCYTE immune cell killing assay.
  • TYK-nu human ovarian cancer cells target cells that naturally expressed p53 R175H and HLA-A2 were labeled with red fluorescent protein (RFP).
  • Fifty thousand effector cells were co-cultured with ten thousand TYK-nu cells over 3 days. The number of viable RFP+ TYK-nu cells were counted at 4h intervals. The results are shown in Figure 1G.
  • EXAMPLE 7 This example demonstrates the specificity and avidity of the 4424-R175H TCR. Effector cells were healthy donor PBL transduced with the retroviral vector encoding the 4424-R175H TCR of Example 4.
  • Target cells were monkey kidney COS7 cells that were engineered to express HLA-A2 and incubated with the WT peptide analogue HMTEVVRRX (SEQ ID NO: 117), wherein X at position 9 of SEQ ID NO: 117 is AABA or Leydig 770310 HHS E-054-2023-0-PC-01 49 the mutant p53 R175H peptide analogue (SEQ ID NO: 116), wherein X at position 9 of SEQ ID NO: 116 is AABA at the concentrations denoted in Fig.2B. Following 18 h co-culture, T cell activation marker 4-1BB expression was measured by flow cytometry. The results are shown in Fig.2B.
  • T cells expressing the 4196-C TCR, 4196-D TCR, 4196-E TCR, or 4424-R175H TCR encoded by the respective retroviral vectors of Example 4 or 4141- TCR1a2 were co-cultured with ten thousand human ovarian cancer TYK-nu cells (target cells) for 72 h at an effector to target ratio of 2:1. Viable RFP+ TYK-nu cells were counted every 3 h. The results are shown in Figure 3. All of the T cell-treated conditions showed a significant delay in tumor growth relative to the tumor-only condition.
  • EXAMPLE 9 This example demonstrates tumor cell recognition by the 4402-Y220C-B TCR, 4402-Y220C-C TCR, and 4402-Y220C-L TCR. Healthy donor PBL were independently transduced with the respective retroviral vectors of Example 4 encoding the 4402-Y220C-B TCR, 4402-Y220C-C TCR, or 4402- Y220C-L TCR (effector cells). Effector cells were co-cultured with target cells under conditions shown in Table 6.
  • the 4402-Y220C-C TCR and 4402-Y220C-L TCR released IFN- ⁇ when the autologous organoid tumor cells were pulsed with p53 Y220C 25-mer (DRNTFRHSVVVPCEPPEVGSDCTTI) (SEQ ID NO: 114).
  • DRNTFRHSVVVPCEPPEVGSDCTTI DRNTFRHSVVVPCEPPEVGSDCTTI
  • the autologous organoid tumor cells exhibited HLA loss of heterozygosity, and they may have lost the HLA-DRB3*02:02 molecule restricting the 4402-Y220C-B TCR.
  • Leydig 770310 HHS E-054-2023-0-PC-01 51 10 This example demonstrates the avidity and specificity of the 4402-Y220C-B TCR, 4402-Y220C-C TCR, and 4402-Y220C-L TCR.
  • Healthy donor PBL were independently transduced with the retroviral vector of Example 4 encoding the 4402-Y220C-B TCR, 4402-Y220C-C TCR, or 4402-Y220C-L TCR or the 4343-D TCR disclosed in US 2023/0321240 (effector cells).
  • Target cells were autologous B cells pulsed with WT (DRNTFRHSVVVPYEPPEVGSDCTTI) (SEQ ID NO: 115) or mutant (DRNTFRHSVVVPCEPPEVGSDCTTI) (SEQ ID NO: 114) peptides at the concentrations denoted in Figures 4A-4D. After the effector cells were co-cultured with the target cells, 4-1BB expression was measured by flow cytometry.
  • Healthy donor PBL were transduced with the retroviral vector of Example 4 encoding the 4402-Y220C-B TCR (effector cells).
  • Target cells were COS7 cells independently transfected with the individual Class II HLA molecules expressed by Patient 4402 (Table 7). TABLE 7 HLA molecule(s) Transfected into Target Cells Leydig 770310 HHS E-054-2023-0-PC-01 52 secretion was measured by ELISpot.
  • 4402-Y220C-B TCR-expressing cells showed significant IFN- ⁇ secretion when co-cultured with COS7 cells expressing HLA-DRB3*02:02.
  • EXAMPLE 12 This example demonstrates differential autologous tumor organoid recognition by 4402-Y220C-C TCR and 4402-Y220C-L TCR. Healthy donor PBL were independently transduced with the retroviral vector of Example 4 encoding the 4402-Y220C-C TCR or 4402-Y220C-L TCR (effector cells).
  • Target cells were autologous breast tumor organoid cells from Patient 4402 of passage 11 or passage 27.
  • Target cells pulsed with DMSO served as a control. Effector cells were co-cultured with target cells. T cell activation was measured by 4-1BB upregulation. The results are shown in Figure 4E.
  • EXAMPLE 13 This example demonstrates the differential expression of the HLA-DRB1*15:01 and DRB1*13:03 molecules by the autologous organoid cells of passage 11 and 27 of Example 12.
  • the expression of HLA-DRB1*15:01 and DRB1*13:03 by the autologous organoid cells of passage 11 and 27 of Example 12 was measured by transcriptome analysis (RNA sequencing). The results are shown in Figure 4F.
  • HLA-DRB1*13:03 is the restriction element for the 4402-Y220C-C TCR and 4402-Y220C-L TCR.
  • Leydig 770310 HHS E-054-2023-0-PC-01 53 14 This example demonstrates that adoptive transfer of cells transduced with the retroviral vector encoding the 4196-C TCR of Example 4 reduce tumor size in tumor-bearing mice.
  • NSG mice (immunocompromised) were subcutaneously injected with TYK-nu human ovarian cancer cells, which naturally expressed the p53 R175H mutation and HLA- A*02:01. Two weeks later, the tumor-bearing mice were randomized and treated with adoptive cell transfer of healthy donor T cells independently transduced with: 1. the retroviral vector encoding the 4196-C TCR of Example 4; 2. an irrelevant TCR control (i.e., 4259); 3. the TCR 4196_AV12-1_with_BV6-1 disclosed in US 2020/0277352; 4. the TCR 4196_AV38-1_with_BV10-3 disclosed in US 2020/0277352; 5.
  • TCR 4196_AV6_with_BV11-2 disclosed in US 2020/0277352; 6. the TCR 4141-TCR1a2 disclosed in US 2022/0332785; or 7. the 4141 IVS TCR disclosed in US Patent Application No.18/289,596.
  • Two T-cell dose levels were tested: 2 million or 10 million T cells/mouse. The mean tumor size was measured at various time points up to 40 days after adoptive transfer of the transduced cells. The results obtained with the 2 million T cells/mouse dose are shown in Figure 6. As shown in Figure 6, only the 4196-C TCR showed statistical significance relative to 4259 TCR, the irrelevant TCR control. The results obtained with the 10 million T cells/mouse dose are shown in Figure 7.
  • Leydig 770310 HHS E-054-2023-0-PC-01 54 followed by a list of one or more items (for “at least one of A and B”) is to be construed to mean one item selected from the listed items (A or B) or any combination of two or more of the listed items (A and B), unless otherwise indicated herein or clearly contradicted by context.
  • the terms “comprising,” “having,” “including,” and “containing” are to be construed as open-ended terms (i.e., meaning “including, but not limited to,”) unless otherwise noted.

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Abstract

Disclosed are isolated or purified T cell receptors (TCRs) having antigenic specificity for human p53Y220C or human p53R175H. Related polypeptides and proteins, as well as related nucleic acids, recombinant expression vectors, host cells, populations of cells, and pharmaceutical compositions are also provided. Also disclosed are methods of detecting the presence of cancer in a mammal and methods of treating or preventing cancer in a mammal.

Description

Leydig 770310 HHS E-054-2023-0-PC-01 1 T CELL RECEPTORS TARGETING Y220C OR R175H MUTATION IN P53 CROSS-REFERENCE TO RELATED APPLICATIONS This patent application claims the benefit of U.S. Provisional Patent Application No.63/492,256, filed March 27, 2023, which is incorporated by reference in its entirety herein. STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT This invention was made with Government support under project number BC010985 by the National Institutes of Health, National Cancer Institute. The Government has certain rights in the invention. INCORPORATION-BY-REFERENCE OF MATERIAL SUBMITTED ELECTRONICALLY Incorporated by reference in its entirety herein is a computer-readable nucleotide/amino acid sequence listing submitted concurrently herewith and identified as follows: One 116,338 Byte XML file named “770310.XML,” dated March 22, 2024. BACKGROUND OF THE INVENTION Some cancers may have very limited treatment options, particularly when the cancer becomes metastatic and unresectable. Despite advances in treatments such as, for example, surgery, chemotherapy, and radiation therapy, the prognosis for many cancers, such as, for example, pancreatic, colorectal, lung, endometrial, ovarian, and prostate cancers, may be poor. Accordingly, there exists an unmet need for additional treatments for cancer. BRIEF SUMMARY OF THE INVENTION An aspect of the invention provides an isolated or purified T cell receptor (TCR) having antigenic specificity for a human p53Y220C or human p53R175H amino acid sequence, wherein the TCR comprises the amino acid sequences of: (1) all of SEQ ID NOs: 2-7; (2) all of SEQ ID NOs: 16-21; (3) all of SEQ ID NOs: 30-35; (4) all of SEQ ID NOs: 44-49; (5) all of SEQ ID NOs: 58-63; (6) all of SEQ ID NOs: 72-77; or (7) all of SEQ ID NOs: 86-91. Leydig 770310 HHS E-054-2023-0-PC-01 2 Further aspects of the invention polypeptides, proteins, nucleic acids, recombinant expression vectors, host cells, populations of cells, and pharmaceutical compositions relating to the TCRs of the invention. Still further aspects of the invention provide methods of detecting the presence of cancer in a mammal, methods of inducing an immune response against a cancer in a mammal, and methods of treating or preventing cancer in a mammal. Additional aspects of the invention provide methods of producing a host cell expressing the TCR and methods of producing the TCR, polypeptide, or protein. BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS Figure 1A is a graph showing the frequency (%) of tetramer positive cells measured following each of culture conditions 1, 2, and 3 of Table 1. Tetramers were generated from HLA-A*02:01 monomer and were loaded with p53 R175H 9-mer peptide analogue to capture p53 R175H mutation-reactive T cells with HLA-A*02:01 restriction. Figures 1B-1E are graphs showing the percentage of mTCR+CD8+ cells expressing 4-1BB following co-culture of effector cells with target cells. Effector cells were healthy donor PBL transduced with 4196-C TCR (B), 4196-D TCR (C), 4196-E TCR (D) or TCR 4196_AV6_with_BV11-2 (E) (disclosed in US 2020/0277352; used as the “Comparison TCR”). Target cells were T2 cells that naturally expressed HLA-A2 and had been incubated with the WT peptide analogue or mutant p53 R175H 9-mer peptide analogue at the indicated concentrations. Figure 1F is a graph including the same data shown in Figures 1B-1E replotted together for comparison. Figure 1G is a graph showing the normalized tumor cell counts measured at 4 hour intervals following co-culture of effector cells with target cells. Effector cells were healthy donor PBL transduced with a retroviral vector encoding 4196-C TCR, 4196-D TCR, 4196-E TCR, or an irrelevant TCR. The target cells were TYK-nu human ovarian cancer cells that naturally expressed p53 R175H and HLA-A*02:01. P<0.001. Figure 2A shows flow cytometry data obtained after staining TILs from Patient 4424 for CD8 and tetramer, which was used in Figure 1A. Figure 2B is a graph showing the percentage of mTCR+CD8+ cells expressing 4- 1BB as a marker for T cell activation following co-culture of effector cells with target cells. Effector cells were healthy donor PBL transduced with the 4424-R175H TCR. Target cells Leydig 770310 HHS E-054-2023-0-PC-01 3 were monkey kidney COS7 cells engineered express HLA-A2 and incubated with the WT peptide analogue or mutant p53 R175H 9-mer peptide analogue at the indicated concentrations. Figure 3 is a graph showing the normalized tumor cell counts measured at 3 hour intervals following co-culture of effector cells with target cells at an effector to target ratio of 2:1. Effector cells were healthy donor PBL transduced with a retroviral vector encoding the 4196-C TCR, 4196-D TCR, 4196-E TCR, 4424-R175H TCR, or 4141-TCR1a2 disclosed in US 2022/0332785 (another “Comparison TCR”). The target cells were TYK-nu human ovarian cancer cells. Statistical testing by 2-way ANOVA. *** p<0.001. Figures 4A-4D are graphs showing the percentage of mTCR+CD3+ cells expressing 4-1BB following co-culture of effector cells with target cells. Effector cells were healthy donor PBL transduced with the 4402-Y220C-B TCR, 4402-Y220C-C TCR, or 4402- Y220C-L TCR or the 4343-D TCR (“Comparison TCR”). Target cells were autologous B cells pulsed with WT or mutant peptide at the indicated concentrations. Figure 4E is a graph showing the percentage of mTCR+CD3+ cells expressing 4- 1BB following co-culture of effector cells with target cells. Effector cells were healthy donor PBL transduced with the 4402-Y220C-C TCR or 4402-Y220C-L TCR. Target cells were autologous breast tumor organoid cells from Patient 4402 of passage 11 showing loss of DRB1*13:03 or passage 27 showing reduced but detectable levels of DRB1*13:03 expression (see Figure 4F). TCR-C and L show recognition of only the organoid passage 27 but not the passage 11, suggesting the two TCRs are restricted by DRB1*13:03. Target cells pulsed with DMSO served as a control. Figure 4F is a graph showing the Transcripts Per Kilobase Million (TPM) value measured for HLA-DRB1*15:01 and DRB1*13:03 in the autologous organoid cells from Patient 4402 of passages 11 and 27. Figures 5A-5C show an alignment of the amino acid sequences of the nine p53 splice variants. SP|P04637|P53_HUMAN (SEQ ID NO: 1); SP|P04637-2|P53_HUMAN (SEQ ID NO: 104); SP|P04637-3|P53_HUMAN (SEQ ID NO: 105); SP|P04637- 4|P53_HUMAN (SEQ ID NO: 106); SP|P04637-5|P53_HUMAN (SEQ ID NO: 107); SP|P04637-6|P53_HUMAN (SEQ ID NO: 108); SP|P04637-7|P53_HUMAN (SEQ ID NO: 109); SP|P04637-8|P53_HUMAN (SEQ ID NO: 110); and SP|P04637-9|P53_HUMAN (SEQ ID NO: 111). The alignment with respect to amino acid residues 1 to 120 of SEQ ID NO: 1 is shown in A, the alignment with respect to amino acid residues 121 to 300 of SEQ ID NO: 1 Leydig 770310 HHS E-054-2023-0-PC-01 4 is shown in B, and the alignment with to amino acid residues 301 to 393 of SEQ ID NO: 1 is shown in C. Figures 6-7 are graphs showing mean tumor size (mm2) measured at the indicated number of days after adoptive cell transfer (ACT) of transduced healthy donor T cells administered to tumor-bearing mice at a dose of 2 million cells/mouse (Figure 6) or 10 million cells/mouse (Figure 7). The healthy donor T cells were independently transduced with: (i) the retroviral vector encoding the 4196-C TCR of Example 4 (“4196 IVS-C”); (ii) an irrelevant TCR control (i.e., 4259) (“4259 (irrelevant)”); (iii) the TCR 4196_AV12- 1_with_BV6-1 disclosed in US 2020/0277352 (“12-6”); (iv) the TCR 4196_AV38- 1_with_BV10-3 disclosed in US 2020/0277352 (“38-10”); (v) the TCR 4196_AV6_with_BV11-2 disclosed in US 2020/0277352 (“6-11”); (vi) the TCR 4141- TCR1a2 disclosed in US 2022/0332785 (“41411a2”); or (vii) the 4141 IVS TCR disclosed in US Patent Application No.18/289,596 (“4141 IVS”). ***Denotes statistical significance. DETAILED DESCRIPTION OF THE INVENTION Tumor Protein P53 (also referred to as “TP53” or “p53”) acts as a tumor suppressor by, for example, regulating cell division. The p53 protein is located in the nucleus of the cell, where it binds directly to DNA. When DNA becomes damaged, the p53 protein is involved in determining whether the DNA will be repaired or the damaged cell will undergo apoptosis. If the DNA can be repaired, p53 activates other genes to fix the damage. If the DNA cannot be repaired, the p53 protein prevents the cell from dividing and signals it to undergo apoptosis. By stopping cells with mutated or damaged DNA from dividing, p53 helps prevent the development of tumors. WT (normal) full-length p53 comprises the amino acid sequence of SEQ ID NO: 1. Mutations in the p53 protein may reduce or eliminate the p53 protein’s tumor suppressor function. Alternatively or additionally, a p53 mutation may be a gain-of-function mutation by interfering with WT p53 in a dominant negative fashion. Mutated p53 protein may be expressed in any of a variety of human cancers such as, for example, cholangiocarcinoma, melanoma, colon cancer, rectal cancer, ovarian cancer, endometrial cancer, non-small cell lung cancer (NSCLC), glioblastoma, uterine cervical cancer, head and neck cancer, breast cancer, pancreatic cancer, or bladder cancer. An aspect of the invention provides an isolated or purified T cell receptor (TCR) having antigenic specificity for a human p53Y220C or human p53R175H amino acid sequence Leydig 770310 HHS E-054-2023-0-PC-01 5 (hereinafter, “mutated p53”). Hereinafter, to a “TCR” also refer to functional portions and functional variants of the TCR, unless specified otherwise. Mutations of p53 are defined herein by reference to the amino acid sequence of full-length, WT p53 (SEQ ID NO: 1). Mutations of p53 are described herein by reference to the amino acid residue present at a particular position, followed by the position number, followed by the amino acid with which that residue has been replaced in the particular mutation under discussion. A p53 amino acid sequence (e.g., a p53 peptide) may comprise fewer than all of the amino acid residues of the full-length, WT p53 protein. Accordingly, the position numbers are defined herein by reference to the WT full-length p53 protein (namely, SEQ ID NO: 1) with the understanding that the actual position of the corresponding residue in a particular example of a p53 amino acid sequence may be different. Because the positions are as defined by SEQ ID NO: 1, the term “Y220C” indicates that the tyrosine present at position 220 of SEQ ID NO: 1 is replaced by cysteine, and “R175H” indicates that the arginine present at position 175 of SEQ ID NO: 1 has been replaced with histidine. For example, when a particular example of a p53 amino acid sequence is, e.g., DRNTFRHSVVVPYEPPEVGSDCTTI (SEQ ID NO: 114) (an exemplary WT p53 peptide corresponding to contiguous amino acid residues 208 to 232 of SEQ ID NO: 1), “Y220C” refers to a substitution of the underlined tyrosine in SEQ ID NO: 114 with cysteine, even though the actual position of the underlined tyrosine in SEQ ID NO: 114 is 13. Human p53 amino acid sequences with the Y220C mutation are hereinafter referred to as “Y220C” or “p53Y220C.” Human p53 amino acid sequences with the R175H mutation are hereinafter referred to as “R175H” or “p53R175H.” As used herein, “mutated p53” refers to human p53Y220C or human p53R175H. P53 has nine known splice variants. The p53 mutations described herein are conserved over all nine p53 splice variants. An alignment of the nine p53 splice variants is shown in Figures 5A-5C. Accordingly, the inventive TCRs may have antigenic specificity for any mutated p53 amino acid sequence described herein encoded by any of the nine p53 splice variants. Because the positions are as defined by SEQ ID NO: 1, then the actual positions of the amino acid sequence of a particular splice variant of p53 are defined relative to the corresponding positions of SEQ ID NO: 1, and the positions as defined by SEQ ID NO: 1 may be different than the actual positions in a particular splice variant. Thus, for example, mutations refer to a replacement of an amino acid residue in the amino acid sequence of a particular splice variant of p53 corresponding to the indicated position of the 393-amino acid Leydig 770310 HHS E-054-2023-0-PC-01 6 sequence of SEQ ID NO: 1 with the that the actual positions in the splice variant may be different. In an aspect of the invention, the TCR has antigenic specificity for human p53 with a mutation at position 220, as defined by SEQ ID NO: 1. The p53 mutation at position 220 may be a missense mutation. Accordingly, the mutation at position 220 may be a substitution of the native (WT) tyrosine residue present at position 220 with any amino acid residue other than tyrosine. In an aspect of the invention, the TCR has antigenic specificity for a human p53Y220C amino acid sequence. For example, the TCR may have antigenic specificity for the human p53Y220C amino acid sequence of DRNTFRHSVVVPCEPPEVGSDCTTI (SEQ ID NO: 114). In an aspect of the invention, the TCR does not have antigenic specificity for the wild-type human p53 amino acid sequence of DRNTFRHSVVVPYEPPEVGSDCTTI (SEQ ID NO: 115). In an aspect of the invention, the TCR has antigenic specificity for human p53 with a mutation at position 175, as defined by SEQ ID NO: 1. The p53 mutation at position 175 may be a missense mutation. Accordingly, the mutation at position 175 may be a substitution of the native (WT) arginine residue present at position 175 with any amino acid residue other than arginine. In an aspect of the invention, the TCR has antigenic specificity for a human p53R175H amino acid sequence. For example, the TCR may have antigenic specificity for the human p53R175H amino acid sequence of HMTEVVRHC (SEQ ID NO: 112). In an aspect of the invention, the TCR does not have antigenic specificity for the wild- type human p53 amino acid sequence of HMTEVVRRC (SEQ ID NO: 113). In an aspect of the invention, the inventive TCRs may be able to recognize mutated p53 in an HLA (human leukocyte antigen)-molecule-dependent manner. “HLA- molecule-dependent manner,” as used herein, means that the TCR elicits an immune response upon binding to mutated p53 within the context of an HLA molecule, which HLA molecule is expressed by the patient from which the TCR was isolated. The inventive TCRs may be able to recognize mutated p53 that is presented by the applicable HLA molecule and may bind to the HLA molecule in addition to mutated p53. In an aspect of the invention, the inventive TCRs are able to recognize Y220C presented by an HLA Class II molecule. In this regard, the TCR may elicit an immune response upon binding to Y220C within the context of an HLA Class II molecule. The inventive TCRs are able to recognize Y220C that is presented by an HLA Class II molecule and may bind to the HLA Class II molecule in addition to Y220C. Leydig 770310 HHS E-054-2023-0-PC-01 7 In an aspect of the invention, the Class II molecule is an HLA-DR heterodimer. The HLA-DR heterodimer is a cell surface receptor including an α chain and a β chain. The HLA-DR α chain is encoded by the HLA-DRA gene. The HLA-DR β chain is encoded by the HLA-DRB1 gene, the HLA-DRB3 gene, HLA-DRB4 gene, or the HLA- DRB5 gene. Examples of molecules encoded by the HLA-DRB1 gene may include, but are not limited to, HLA-DR1, HLA-DR2, HLA-DR3, HLA-DR4, HLA-DR5, HLA-DR6, HLA- DR7, HLA-DR8, HLA-DR9, HLA-DR10, HLA-DR11, HLA-DR12, HLA-DR13, HLA- DR14, HLA-DR15, HLA-DR16, and HLA-DR17. The HLA-DRB3 gene encodes HLA- DR52. The HLA-DRB4 gene encodes HLA-DR53. The HLA-DRB5 gene encodes HLA- DR51. In an aspect, the alpha chain of the HLA Class II molecule is expressed by the HLA-DRA1*01:01 allele. In an aspect, the beta chain of the HLA Class II molecule is expressed by the HLA-DRB3*02:02 allele. In an aspect of the invention, the HLA Class II molecule is an HLA-DRB3:HLA-DRA heterodimer. In a preferred aspect, the HLA Class II molecule is a heterodimer of an HLA-DRA1*01:01 chain and an HLA-DRB3*02:02 chain. In an especially preferred aspect, the mutated p53 is Y220C and the HLA Class II molecule is a heterodimer of an HLA-DRA1*01:01 chain and an HLA-DRB3*02:02 chain. In an aspect, the alpha chain of the HLA Class II molecule is expressed by the HLA-DRA1*01:01 allele. In an aspect, the beta chain of the HLA Class II molecule is expressed by the HLA-DRB1*13:03 allele. In an aspect of the invention, the HLA Class II molecule is an HLA-DRB1:HLA-DRA heterodimer. In a preferred aspect, the HLA Class II molecule is a heterodimer of an HLA-DRA1*01:01 chain and an HLA-DRB1*13:03 chain. In an especially preferred aspect, the mutated p53 is Y220C and the HLA Class II molecule is a heterodimer of an HLA-DRA1*01:01 chain and an HLA-DRB1*13:03 chain. In an aspect of the invention, the inventive TCRs are able to recognize R175H presented by an HLA Class I molecule. In this regard, the TCR may elicit an immune response upon binding to R175H within the context of an HLA Class I molecule. The inventive TCRs are able to recognize R175H that is presented by an HLA Class I molecule and may bind to the HLA Class I molecule in addition to R175H. In an embodiment of the invention, the HLA Class I molecule is an HLA-A molecule. The HLA-A molecule is a heterodimer of an α chain and β2 microglobulin. The HLA-A α chain may be encoded by an HLA-A gene. β2 microglobulin binds non-covalently to the alpha1, alpha2 and alpha3 domains of the alpha chain to build the HLA-A complex. Leydig 770310 HHS E-054-2023-0-PC-01 8 The HLA-A molecule may be any HLA-A In an embodiment of the invention, the HLA Class I molecule is an HLA-A2 molecule. The HLA-A2 molecule may be any HLA- A2 molecule. Examples of HLA-A2 molecules may include, but are not limited to, those encoded by the HLA-A*02:01, HLA-A*02:02, HLA-A*02:03 allele, HLA-A*02:05, HLA- A*02:06, HLA-A*02:07 allele, or HLA-A*02:11 allele. Preferably, the HLA Class I molecule is encoded by the HLA-A*02:01 allele. The TCRs of the invention may provide any one or more of many advantages, including when expressed by cells used for adoptive cell transfer. Mutated p53 is expressed by cancer cells and is not expressed by normal, noncancerous cells. Without being bound to a particular theory or mechanism, it is believed that the inventive TCRs advantageously target the destruction of cancer cells while minimizing or eliminating the destruction of normal, non-cancerous cells, thereby reducing, for example, by minimizing or eliminating, toxicity. Moreover, the inventive TCRs may, advantageously, successfully treat or prevent mutated p53-positive cancers that do not respond to other types of treatment such as, for example, chemotherapy, surgery, or radiation. Additionally, the inventive TCRs may provide highly avid recognition of mutated p53, which may provide the ability to recognize unmanipulated tumor cells (e.g., tumor cells that have not been treated with interferon (IFN)-γ, transfected with a vector encoding one or both of mutated p53 and the applicable HLA molecule, pulsed with a p53 peptide with the p53 mutation, or a combination thereof). Mutations in p53 are common across different tumor types. Roughly half of all tumors harbor a mutation in p53, about half of which will be a missense mutation. The R175H mutation is common, affecting about 5% of all patients with solid cancers. The Y220C mutation is also highly recurrent and affects about 0.4% of all cancer patients. Accordingly, the inventive TCRs may increase the number of patients who may be eligible for treatment with immunotherapy. The phrase “antigenic specificity,” as used herein, means that the TCR can specifically bind to and immunologically recognize mutated p53 with high avidity. For example, a TCR may be considered to have “antigenic specificity” for mutated p53 if about 1 x 104 to about 1 x 105 T cells expressing the TCR secrete at least about 200 pg/mL or more (e.g., 200 pg/mL or more, 300 pg/mL or more, 400 pg/mL or more, 500 pg/mL or more, 600 pg/mL or more, 700 pg/mL or more, 1000 pg/mL or more, 5,000 pg/mL or more, 7,000 pg/mL or more, 10,000 pg/mL or more, 20,000 pg/mL or more, or a range defined by any two of the foregoing values) of IFN-γ upon co-culture with (a) antigen-negative, applicable HLA molecule positive target cells pulsed with mutated p53 peptide (e.g., about 0.1 ng/mL to Leydig 770310 HHS E-054-2023-0-PC-01 9 about 10,000 ng/mL, 0.1 ng/mL, 0.5 ng/mL, ng/mL, 5 ng/mL, 10 ng/mL, 100 ng/mL, 500 ng/mL, 1,000 ng/mL, 5,000 ng/mL, 10,000 ng/mL, or a range defined by any two of the foregoing values) or (b) antigen-negative, applicable HLA molecule positive target cells into which a nucleotide sequence encoding mutated p53 has been introduced such that the target cell expresses mutated p53. Cells expressing the inventive TCRs may also secrete IFN-γ upon co-culture with antigen-negative, applicable HLA molecule positive target cells pulsed with higher concentrations of mutated p53 peptide. Alternatively or additionally, a TCR may be considered to have “antigenic specificity” for mutated p53 if T cells expressing the TCR secrete at least twice as much IFN- γ upon co-culture with (a) antigen-negative, applicable HLA molecule positive target cells pulsed with mutated p53 peptide or (b) antigen-negative, applicable HLA molecule positive target cells into which a nucleotide sequence encoding mutated p53 has been introduced such that the target cell expresses mutated p53 as compared to the amount of IFN-γ expressed by a negative control. The negative control may be, for example, (i) T cells expressing the TCR, co-cultured with (a) antigen-negative, applicable HLA molecule positive target cells pulsed with the same concentration of an irrelevant peptide (e.g., some other peptide with a different sequence from the mutated p53 peptide) or (b) antigen-negative, applicable HLA molecule positive target cells into which a nucleotide sequence encoding an irrelevant peptide has been introduced such that the target cell expresses the irrelevant peptide, or (ii) untransduced T cells (e.g., derived from PBMC, which do not express the TCR) co-cultured with (a) antigen- negative, applicable HLA molecule positive target cells pulsed with the same concentration of mutated p53 peptide or (b) antigen-negative, applicable HLA molecule positive target cells into which a nucleotide sequence encoding mutated p53 has been introduced such that the target cell expresses mutated p53. IFN-γ secretion may be measured by methods known in the art such as, for example, enzyme-linked immunosorbent assay (ELISA). The concentration of pulsed peptide may be as described herein with respect to other aspects of the invention. Alternatively or additionally, a TCR may be considered to have “antigenic specificity” for mutated p53 if at least twice as many of the numbers of T cells expressing the TCR secrete IFN-γ upon co-culture with (a) antigen-negative, applicable HLA molecule positive target cells pulsed with mutated p53 peptide or (b) antigen-negative, applicable HLA molecule positive target cells into which a nucleotide sequence encoding mutated p53 has been introduced such that the target cell expresses mutated p53 as compared to the numbers Leydig 770310 HHS E-054-2023-0-PC-01 10 of negative control T cells that secrete γ. The concentration of peptide and the negative control may be as described herein with respect to other aspects of the invention. The numbers of cells secreting IFN-γ may be measured by methods known in the art such as, for example, enzyme-linked immunospot (ELISPOT) assay. Alternatively or additionally, a TCR may be considered to have “antigenic specificity” for mutated p53 if at least twice as many spots are detected by ELISPOT for the T cells expressing the TCR upon co-culture with (a) antigen-negative, applicable HLA molecule positive target cells pulsed with mutated p53 peptide or (b) antigen-negative, applicable HLA molecule positive target cells into which a nucleotide sequence encoding mutated p53 has been introduced such that the target cell expresses mutated p53 as compared to the number of spots detected by ELISPOT for negative control T cells co-cultured with the same target cells. The concentration of peptide and the negative control may be as described herein with respect to other aspects of the invention. Alternatively or additionally, a TCR may be considered to have “antigenic specificity” for mutated p53 if T cells expressing the TCR upregulate expression of one or both of 4-1BB and OX40 as measured by, for example, flow cytometry after stimulation with target cells expressing mutated p53. An aspect of the invention provides a TCR comprising two polypeptides (i.e., polypeptide chains), such as an alpha (α) chain of a TCR, a beta (β) chain of a TCR, a gamma (γ) chain of a TCR, a delta (δ) chain of a TCR, or a combination thereof. The polypeptides of the inventive TCR can comprise any amino acid sequence, provided that the TCR has antigenic specificity for mutated p53. In an aspect of the invention, the TCR comprises two polypeptide chains, each of which comprises a variable region comprising a complementarity determining region (CDR)1, a CDR2, and a CDR3 of a TCR. In an aspect of the invention, the TCR comprises a first polypeptide chain comprising an α chain CDR1 (CDR1α), an α chain CDR2 (CDR2α), and an α chain CDR3 (CDR3α), and a second polypeptide chain comprising a β chain CDR1 (CDR1β), a β chain CDR2 (CDR2β), and a β chain CDR3 (CDR3β). In an aspect of the invention, the TCR comprises the amino acid sequences of: (1) all of SEQ ID NOs: 2-7 (4196-C TCR); (2) all of SEQ ID NOs: 16-21 (4196-D TCR); (3) all of SEQ ID NOs: 30-35 (4196-E TCR); (4) all of SEQ ID NOs: 44-49 (4424-R175H TCR); (5) all of SEQ ID NOs: 58-63 (4402-Y220C-B TCR); (6) all of SEQ ID NOs: 72-77 (4402-Y220C-C TCR); or (7) all of SEQ ID NOs: 86-91 (4402-Y220C-L TCR). Each one of the foregoing seven collections Leydig 770310 HHS E-054-2023-0-PC-01 11 of amino acid sequences in this paragraph forth the six CDR regions of each of seven different TCRs having antigenic specificity for mutated human p53. The six amino acid sequences in each collection correspond to the CDR1α, CDR2α, CDR3α, CDR1β, CDR2β, and CDR3β of a TCR, respectively. The TCR may comprise the amino acid sequences of any one or more of: SEQ ID NOs: 2-7, 16-21, 30-35, 44-49, 58-63, 72-77, and 86-91. In an aspect of the invention, the TCR comprises an isolated or purified T cell receptor (TCR) having antigenic specificity for a human p53Y220C or human p53R175H amino acid sequence, wherein the TCR comprises the amino acid sequences of: (1) all of SEQ ID NOs: 2-7; (2) all of SEQ ID NOs: 16-21; (3) all of SEQ ID NOs: 30-35; (4) all of SEQ ID NOs: 44-49; (5) all of SEQ ID NOs: 58-63; (6) all of SEQ ID NOs: 72-77; or (7) all of SEQ ID NOs: 86-91. In an aspect of the invention, the TCR comprises an α chain variable region amino acid sequence and a β chain variable region amino acid sequence, which together comprise one of the collections of CDRs set forth above. In this regard, the TCR can comprise the amino acid sequences of: (1) both of SEQ ID NOs: 8 and 9 (4196-C TCR); (2) both of SEQ ID NOs: 10 and 11 (4196-C TCR); (3) both of SEQ ID NOs: 22 and 23 (4196-D TCR); (4) both of SEQ ID NOs: 24 and 25 (4196-D TCR); (5) both of SEQ ID NOs: 36 and 37 (4196-E TCR); (6) both of SEQ ID NOs: 38 and 39 (4196-E TCR); (7) both of SEQ ID NOs: 50 and 51 (4424-R175H TCR); (8) both of SEQ ID NOs: 52 and 53 (4424-R175H TCR); (9) both of SEQ ID NOs: 64 and 65 (4402-Y220C-B TCR); (10) both of SEQ ID NOs: 66 and 67 (4402- Y220C-B TCR); (11) SEQ ID NO: 78 and 79 (4402-Y220C-C TCR); (12) SEQ ID NO: 80 and 81 (4402-Y220C-C TCR); (13) SEQ ID NO: 92 and 93 (4402-Y220C-L TCR); or (14) SEQ ID NO: 94 and 95 (4402-Y220C-L TCR). Each one of the foregoing collections of amino acid sequences in this paragraph sets forth the two variable regions of each of seven different TCRs having antigenic specificity for mutated human p53. The two amino acid sequences in each collection correspond to the variable region of the α chain and the variable region of the β chain of a TCR, respectively. The TCR may, e.g., comprise the amino acid sequence of any one or more of SEQ ID NOs: 8, 9, 10, 11, 22, 23, 24, 25, 36, 37, 38, 39, 50, 51, 52, 53, 64, 65, 66, 67, 78, 79, 80, 81, 92, 93, 94, and 95. In an aspect of the invention, the TCR comprises the amino acid sequence(s) of: (1) SEQ ID NO: 8; (2) SEQ ID NO: 9; (3) both of SEQ ID NOs: 8 and 9; (4) SEQ ID NO: 10; (5) SEQ ID NO: 11; (6) both of SEQ ID NOs: 10 and 11; (7) SEQ ID NO: 22; (8) SEQ ID NO: 23; (9) both of SEQ ID NOs: 22 and 23; (10) SEQ ID NO: 24; (11) SEQ Leydig 770310 HHS E-054-2023-0-PC-01 12 ID NO: 25; (12) both of SEQ ID NOs: 24 (13) SEQ ID NO: 36; (14) SEQ ID NOs: 37; (15) both of SEQ ID NOs: 36 and 37; (16) SEQ ID NO: 38; (17) SEQ ID NO: 39; (18) both of SEQ ID NOs: 38 and 39; (19) SEQ ID NO: 50; (20) SEQ ID NO: 51; (21) both of SEQ ID NOs: 50 and 51; (22) SEQ ID NO: 52; (23) SEQ ID NO: 53; (24) both of SEQ ID NOs: 52 and 53; (25) SEQ ID NO: 64; (26) SEQ ID NO: 65; (27) both of SEQ ID NOs: 64 and 65; (28) SEQ ID NO: 66; (29) SEQ ID NO: 67; (30) both of SEQ ID NOs: 66 and 67; (31) SEQ ID NO: 78; (32) SEQ ID NO: 79; (33) both of SEQ ID NOs: 78 and 79; (34) SEQ ID NO: 80; (35) SEQ ID NO: 81; (36) both of SEQ ID NOs: 80 and 81; (37) SEQ ID NO: 92; (38) SEQ ID NO: 93; (39) both of SEQ ID NOs: 92 and 93; (40) SEQ ID NO: 94; (41) SEQ ID NO: 95; or (42) both of SEQ ID NO: 94 and 95. The inventive TCRs may further comprise a constant region. The constant region may be derived from any suitable species such as, e.g., human or mouse. In an aspect of the invention, the TCRs further comprise a murine constant region. As used herein, the term “murine” or “human,” when referring to a TCR or any component of a TCR described herein (e.g., complementarity determining region (CDR), variable region, constant region, alpha chain, and/or beta chain), means a TCR (or component thereof) which is derived from a mouse or a human, respectively, i.e., a TCR (or component thereof) that originated from or was, at one time, expressed by a mouse T cell or a human T cell, respectively. In an aspect of the invention, the TCR may comprise a murine α chain constant region and a murine β chain constant region. The murine α chain constant region may be modified or unmodified. A modified murine α chain constant region may be, e.g., cysteine-substituted, LVL-modified, or both cysteine-substituted and LVL-modified, as described, for example, in U.S. Patent No. 10,174,098. The murine β chain constant region may be modified or unmodified. A modified murine β chain constant region may be, e.g., cysteine-substituted, as described, for example, in U.S. Patent No.10,174,098. In an aspect of the invention, the TCR comprises a cysteine-substituted, LVL-modified murine α chain constant region comprising the amino acid sequence of SEQ ID NO: 100 or 101. In an aspect of the invention, the TCR comprises a cysteine-substituted murine β chain constant region comprising the amino acid sequence of SEQ ID NO: 102. In an aspect of the invention, the inventive TCR can comprise an α chain of a TCR and a β chain of a TCR. The α chain of the TCR may comprise a variable region of an α chain and a constant region of an α chain. An α chain of this type can be paired with any β Leydig 770310 HHS E-054-2023-0-PC-01 13 chain of a TCR. The β chain may a region of a β chain and a constant region of a β chain. In an aspect of the invention, the TCR can comprise the amino acid sequences of: (1) both of SEQ ID NOs: 12 and 13 (4196-C TCR); (2) both of SEQ ID NOs: 14 and 15 (4196-C TCR); (3) both of SEQ ID NOs: 26 and 27 (4196-D TCR); (4) both of SEQ ID NOs: 28 and 29 (4196-D TCR); (5) both of SEQ ID NOs: 40 and 41 (4196-E TCR); (6) both of SEQ ID NOs: 42 and 43 (4196-E TCR); (7) both of SEQ ID NOs: 54 and 55 (4424-R175H TCR); (8) both of SEQ ID NOs: 56 and 57 (4424-R175H); (9) both of SEQ ID NOs: 68 and 69 (4402-Y220C-B TCR); (10) both of SEQ ID NOs: 70 and 71 (4402-Y220C-B TCR); (11) SEQ ID NO: 82 and 83 (4402-Y220C-C TCR); (12) SEQ ID NO: 84 and 85 (4402-Y220C-C TCR); (13) SEQ ID NO: 96 and 97 (4402-Y220C-L TCR); or (14) SEQ ID NO: 98 and 99 (4402-Y220C-L TCR). Each one of the foregoing collections of amino acid sequences in this paragraph sets forth the α chain and β chain of each of seven different TCRs having antigenic specificity for mutated human p53. The two amino acid sequences in each collection correspond to the α chain and the β chain of a TCR, respectively. The TCR may comprise the amino acid sequence of any one or more of SEQ ID NOs: 12, 13, 14, 15, 26, 27, 28, 29, 40, 41, 42, 43, 54, 55, 56, 57, 68, 69, 70, 71, 82, 83, 84, 85, 96, 97, 98, and 99. In an aspect of the invention, the TCR comprises the amino acid sequences of: (1) SEQ ID NO: 12; (2) SEQ ID NO: 13; (3) both of SEQ ID NOs: 12 and 13; (4) SEQ ID NO: 14; (5) SEQ ID NO: 15; (6) both of SEQ ID NOs: 14 and 15; (7) SEQ ID NO: 26; (8) SEQ ID NO: 27; (9) both of SEQ ID NOs: 26 and 27; (10) SEQ ID NO: 28; (11) SEQ ID NO: 29; (12) both of SEQ ID NOs: 28 and 29; (13) SEQ ID NO: 40; (14) SEQ ID NO: 41; (15) both of SEQ ID NOs: 40 and 41; (16) SEQ ID NO: 42; (17) SEQ ID NO: 43; (18) both of SEQ ID NOs: 42 and 43; (19) SEQ ID NO: 54; (20) SEQ ID NO: 55; (21) both of SEQ ID NOs: 54 and 55; (22) SEQ ID NO: 56; (23) SEQ ID NO: 57; (24) both of SEQ ID NOs: 56 and 57; (25) SEQ ID NO: 68; (26) SEQ ID NO: 69; (27) both of SEQ ID NOs: 68 and 69; (28) SEQ ID NO: 70; (29) SEQ ID NO: 71; (30) both of SEQ ID NOs: 70 and 71; (31) SEQ ID NO: 82; (32) SEQ ID NO: 83; (33) both of SEQ ID NO: 82 and 83; (34) SEQ ID NO: 84; (35) SEQ ID NO: 85; (36) both of SEQ ID NO: 84 and 85; (37) SEQ ID NO: 96; (38) SEQ ID NO: 97; (39) both of SEQ ID NO: 96 and 97; (40) SEQ ID NO: 98; (41) SEQ ID NO: 99; or (42) both of SEQ ID NO: 98 and 99. Included in the scope of the invention are functional variants of the inventive TCRs described herein. The term “functional variant,” as used herein, refers to a TCR, Leydig 770310 HHS E-054-2023-0-PC-01 14 polypeptide, or protein having substantial or sequence identity or similarity to a parent TCR, polypeptide, or protein, which functional variant retains the biological activity of the TCR, polypeptide, or protein of which it is a variant. Functional variants encompass, for example, those variants of the TCR, polypeptide, or protein described herein (the parent TCR, polypeptide, or protein) that retain the ability to specifically bind to mutated p53 for which the parent TCR has antigenic specificity or to which the parent polypeptide or protein specifically binds, to a similar extent, the same extent, or to a higher extent, as the parent TCR, polypeptide, or protein. In reference to the parent TCR, polypeptide, or protein, the functional variant can, for instance, be at least about 30%, at least about 50%, at least about 75%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or more identical in amino acid sequence to the parent TCR, polypeptide, or protein, respectively. The functional variant can, for example, comprise the amino acid sequence of the parent TCR, polypeptide, or protein with at least one conservative amino acid substitution. Conservative amino acid substitutions are known in the art, and include amino acid substitutions in which one amino acid having certain physical and/or chemical properties is exchanged for another amino acid that has the same chemical or physical properties. For instance, the conservative amino acid substitution can be an acidic amino acid substituted for another acidic amino acid (e.g., Asp or Glu), an amino acid with a nonpolar side chain substituted for another amino acid with a nonpolar side chain (e.g., Ala, Gly, Val, Ile, Leu, Met, Phe, Pro, Trp, Val, etc.), a basic amino acid substituted for another basic amino acid (Lys, Arg, etc.), an amino acid with a polar side chain substituted for another amino acid with a polar side chain (Asn, Cys, Gln, Ser, Thr, Tyr, etc.), etc. Alternatively or additionally, the functional variants can comprise the amino acid sequence of the parent TCR, polypeptide, or protein with at least one non-conservative amino acid substitution. In this case, it is preferable for the non-conservative amino acid substitution to not interfere with or inhibit the biological activity of the functional variant. Preferably, the non-conservative amino acid substitution enhances the biological activity of the functional variant, such that the biological activity of the functional variant is increased as compared to the parent TCR, polypeptide, or protein. The TCR, polypeptide, or protein can consist essentially of the specified amino acid sequence or sequences described herein, such that other components of the TCR, Leydig 770310 HHS E-054-2023-0-PC-01 15 polypeptide, or protein, e.g., other amino do not materially change the biological activity of the TCR, polypeptide, or protein. Also provided by the invention is a polypeptide comprising a functional portion of any of the TCRs described herein. The term “polypeptide,” as used herein, includes oligopeptides and refers to a single chain of amino acids connected by one or more peptide bonds. With respect to the inventive polypeptides, the functional portion can be any portion comprising contiguous amino acids of the TCR of which it is a part, provided that the functional portion specifically binds to mutated p53. The term “functional portion,” when used in reference to a TCR, refers to any part or fragment of the TCR of the invention, which part or fragment retains the biological activity of the TCR of which it is a part (the parent TCR). Functional portions encompass, for example, those parts of a TCR that retain the ability to specifically bind to mutated p53 (e.g., in an applicable HLA molecule-dependent manner), or detect, treat, or prevent cancer, to a similar extent, the same extent, or to a higher extent, as the parent TCR. In reference to the parent TCR, the functional portion can comprise, for instance, about 10%, about 25%, about 30%, about 50%, about 70%, about 80%, about 90%, about 95%, or more, of the parent TCR. The functional portion can comprise additional amino acids at the amino or carboxy terminus of the portion, or at both termini, which additional amino acids are not found in the amino acid sequence of the parent TCR. Desirably, the additional amino acids do not interfere with the biological function of the functional portion, e.g., specifically binding to mutated p53; and/or having the ability to detect cancer, treat or prevent cancer, etc. More desirably, the additional amino acids enhance the biological activity, as compared to the biological activity of the parent TCR. The polypeptide can comprise a functional portion of either or both of the α and β chains of the TCRs of the invention, such as a functional portion comprising one of more of CDR1, CDR2, and CDR3 of the variable region(s) of the α chain and/or β chain of a TCR of the invention. In an aspect of the invention, the polypeptide comprises the amino acid sequences of: (1) all of SEQ ID NOs: 2-7; (2) all of SEQ ID NOs: 16-21; (3) all of SEQ ID NOs: 30-35; (4) all of SEQ ID NOs: 44-49; (5) all of SEQ ID NOs: 58-63; (6) all of SEQ ID NOs: 72-77; or (7) all of SEQ ID NOs: 86-91. The polypeptide may comprise the amino acid sequences of any one or more of: SEQ ID NOs: 2-7, 16-21, 30-35, 44-49, 58-63, 72-77, and 86-91. Leydig 770310 HHS E-054-2023-0-PC-01 16 In an aspect of the invention, polypeptide can comprise, for instance, the variable region of the inventive TCR comprising a combination of the CDR regions set forth above. In this regard, the polypeptide can comprise, e.g., the amino acid sequences of: (1) both of SEQ ID NOs: 8 and 9; (2) both of SEQ ID NOs: 10 and 11; (3) both of SEQ ID NOs: 22 and 23; (4) both of SEQ ID NOs: 24 and 25; (5) both of SEQ ID NOs: 36 and 37; (6) both of SEQ ID NOs: 38 and 39; (7) both of SEQ ID NOs: 50 and 51; (8) both of SEQ ID NOs: 52 and 53; (9) both of SEQ ID NOs: 64 and 65; (10) both of SEQ ID NOs: 66 and 67; (11) both of SEQ ID NOs: 78 and 79; (12) both of SEQ ID NOs: 80 and 81; (13) both of SEQ ID NOs: 92 and 93; or (14) both of SEQ ID NOs: 94 and 95. The polypeptide may, e.g., comprise the amino acid sequence of any one or more of SEQ ID NOs: 8, 9, 10, 11, 22, 23, 24, 25, 36, 37, 38, 39, 50, 51, 52, 53, 64, 65, 66, 67, 78, 79, 80, 81, 92, 93, 94 and 95. In an aspect of the invention, the polypeptide comprises the amino acid sequence(s) of: (1) SEQ ID NO: 8; (2) SEQ ID NO: 9; (3) both of SEQ ID NOs: 8 and 9; (4) SEQ ID NO: 10; (5) SEQ ID NO: 11; (6) both of SEQ ID NOs: 10 and 11; (7) SEQ ID NO: 22; (8) SEQ ID NO: 23; (9) both of SEQ ID NOs: 22 and 23; (10) SEQ ID NO: 24; (11) SEQ ID NO: 25; (12) both of SEQ ID NOs: 24 and 25; (13) SEQ ID NO: 36; (14) SEQ ID NOs: 37; (15) both of SEQ ID NOs: 36 and 37; (16) SEQ ID NO: 38; (17) SEQ ID NO: 39; (18) both of SEQ ID NOs: 38 and 39; (19) SEQ ID NO: 50; (20) SEQ ID NO: 51; (21) both of SEQ ID NOs: 50 and 51; (22) SEQ ID NO: 52; (23) SEQ ID NO: 53; (24) both of SEQ ID NOs: 52 and 53; (25) SEQ ID NO: 64; (26) SEQ ID NO: 65; (27) both of SEQ ID NOs: 64 and 65; (28) SEQ ID NO: 66; (29) SEQ ID NO: 67; (30) both of SEQ ID NOs: 66 and 67; (31) SEQ ID NO: 78; (32) SEQ ID NO: 79; (33) both of SEQ ID NOs: 78 and 79; (34) SEQ ID NO: 80; (35) SEQ ID NO: 81; (36) both of SEQ ID NOs: 80 and 81; (37) SEQ ID NO: 92; (38) SEQ ID NO: 93; (39) both of SEQ ID NOs: 92 and 93; (40) SEQ ID NO: 94; (41) SEQ ID NO: 95; or (42) both of SEQ ID NO: 94 and 95. In an aspect of the invention, the inventive polypeptide can further comprise the constant region of the inventive TCR set forth above. In this regard, the polypeptide can comprise, e.g., the amino acid sequence of (i) one of SEQ ID NOs 100-102 or (ii) SEQ ID NO: 102 and one of SEQ ID NOs: 100 and 101. In an aspect of the invention, the inventive polypeptide may comprise an α chain and a β chain of the inventive TCR. In this regard, the polypeptide can comprise, e.g., the amino acid sequences of: (1) both of SEQ ID NOs: 12 and 13; (2) both of SEQ ID NOs: 14 and 15; (3) both of SEQ ID NOs: 26 and 27; (4) both of SEQ ID NOs: 28 and 29; (5) both of Leydig 770310 HHS E-054-2023-0-PC-01 17 SEQ ID NOs: 40 and 41; (6) both of SEQ 42 and 43; (7) both of SEQ ID NOs: 54 and 55; (8) both of SEQ ID NOs: 56 and 57; (9) both of SEQ ID NOs: 68 and 69; (10) both of SEQ ID NOs: 70 and 71; (11) SEQ ID NO: 82 and 83; (12) SEQ ID NO: 84 and 85; (13) SEQ ID NO: 96 and 97; or (14) SEQ ID NO: 98 and 99. The polypeptide may comprise the amino acid sequence of any one or more of SEQ ID NOs: 12, 13, 14, 15, 26, 27, 28, 29, 40, 41, 42, 43, 54, 55, 56, 57, 68, 69, 70, 71, 82, 83, 84, 85, 96, 97, 98, and 99. In an aspect of the invention, the polypeptide comprises the amino acid sequences of: (1) SEQ ID NO: 12; (2) SEQ ID NO: 13; (3) both of SEQ ID NOs: 12 and 13; (4) SEQ ID NO: 14; (5) SEQ ID NO: 15; (6) both of SEQ ID NOs: 14 and 15; (7) SEQ ID NO: 26; (8) SEQ ID NO: 27; (9) both of SEQ ID NOs: 26 and 27; (10) SEQ ID NO: 28; (11) SEQ ID NO: 29; (12) both of SEQ ID NOs: 28 and 29; (13) SEQ ID NO: 40; (14) SEQ ID NO: 41; (15) both of SEQ ID NOs: 40 and 41; (16) SEQ ID NO: 42; (17) SEQ ID NO: 43; (18) both of SEQ ID NOs: 42 and 43; (19) SEQ ID NO: 54; (20) SEQ ID NO: 55; (21) both of SEQ ID NOs: 54 and 55; (22) SEQ ID NO: 56; (23) SEQ ID NO: 57; (24) both of SEQ ID NOs: 56 and 57; (25) SEQ ID NO: 68; (26) SEQ ID NO: 69; (27) both of SEQ ID NOs: 68 and 69; (28) SEQ ID NO: 70; (29) SEQ ID NO: 71; (30) both of SEQ ID NOs: 70 and 71; (31) SEQ ID NO: 82; (32) SEQ ID NO: 83; (33) both of SEQ ID NO: 82 and 83; (34) SEQ ID NO: 84; (35) SEQ ID NO: 85; (36) both of SEQ ID NO: 84 and 85; (37) SEQ ID NO: 96; (38) SEQ ID NO: 97; (39) both of SEQ ID NO: 96 and 97; (40) SEQ ID NO: 98; (41) SEQ ID NO: 99; or (42) both of SEQ ID NO: 98 and 99. An aspect of the invention further provides a protein comprising a functional portion of any of the inventive TCRs described herein. By “protein” is meant a molecule comprising one or more polypeptide chains. In an aspect, the protein of the invention can comprise: first and second polypeptide chains, wherein: (1) the first polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 2-4; (2) the second polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 5-7; (3) the first polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 2-4 and the second polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 5-7; (4) the first polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 16-18; (5) the second polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 19-21; (6) the first polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 16-18 and the second polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 19-21; (7) the first polypeptide chain comprises the amino acid sequences of Leydig 770310 HHS E-054-2023-0-PC-01 18 all of SEQ ID NOs: 30-32; (8) the second chain comprises the amino acid sequences of all of SEQ ID NOs: 33-35; (9) the first polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 30-32 and the second polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 33-35; (10) the first polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 44-46; (11) the second polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 47-49; (12) the first polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 44-46 and the second polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 47- 49; (13) the first polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 58-60; (14) the second polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 61-63; (15) the first polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 58-60 and the second polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 61-63; (16) the first polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 72-74; (17) the second polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 75-77; (18) the first polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 72-74 and the second polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 75-77; (19) the first polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 86-88; (20) the second polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 89-91; or (21) the first polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 86-88 and the second polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 89-91. In an aspect of the invention, the protein comprises first and second polypeptide chains, wherein: (1) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 8; (2) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 9; (3) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 8 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 9; (4) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 10; (5) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 11; (6) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 10 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 11; (7) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 22; (8) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 23; (9) the first Leydig 770310 HHS E-054-2023-0-PC-01 19 polypeptide chain comprises the amino acid sequence of SEQ ID NO: 22 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 23; (10) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 24; (11) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 25; (12) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 24 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 25; (13) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 36; (14) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 37; (15) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 36 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 37; (16) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 38; (17) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 39; (18) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 38 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 39; (19) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 50; (20) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 51; (21) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 50 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 51; (22) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 52; (23) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 53; (24) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 52 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 53; (25) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 64; (26) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 65; (27) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 64 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 65; (28) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 66; (29) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 67; (30) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 66 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 67; (31) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 78; (32) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 79; (33) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 78 and the second Leydig 770310 HHS E-054-2023-0-PC-01 20 polypeptide chain comprises the amino acid sequence of SEQ ID NO: 79; (34) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 80; (35) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 81; (36) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 80 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 81; (37) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 92; (38) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 93; (39) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 92 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 93; (40) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 94; (41) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 95; or (42) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 94 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 95. In an aspect of the invention, the protein comprises first and second polypeptide chains, wherein: (1) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 12; (2) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 13; (3) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 12 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 13; (4) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 14; (5) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 15; (6) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 14 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 15; (7) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 26; (8) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 27; (9) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 26 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 27; (10) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 28; (11) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 29; (12) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 28 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 29; (13) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 40; (14) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 41; (15) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 40 and the second Leydig 770310 HHS E-054-2023-0-PC-01 21 polypeptide chain comprises the amino acid sequence of SEQ ID NO: 41; (16) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 42; (17) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 43; (18) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 42 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 43; (19) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 54; (20) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 55; (21) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 54 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 55; (22) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 56; (23) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 57; (24) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 56 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 57; (25) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 68; (26) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 69; (27) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 68 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 69; (28) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 70; (29) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 71; (30) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 70 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 71; (31) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 82; (32) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 83; (33) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 82 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 83; (34) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 84; (35) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 85; (36) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 84 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 85; (37) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 96; (38) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 97; (39) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 96 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 97; (40) the first Leydig 770310 HHS E-054-2023-0-PC-01 22 polypeptide chain comprises the amino acid sequence of SEQ ID NO: 98; (41) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 99; or (42) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 98 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 99. The protein of the invention may be a TCR. Alternatively, if the first and/or second polypeptide chain(s) of the protein further comprise(s) other amino acid sequences, e.g., an amino acid sequence encoding an immunoglobulin or a portion thereof, then the inventive protein can be a fusion protein. In this regard, an aspect of the invention also provides a fusion protein comprising at least one of the inventive polypeptides described herein along with at least one other polypeptide. The other polypeptide can exist as a separate polypeptide of the fusion protein, or can exist as a polypeptide, which is expressed in frame (in tandem) with one of the inventive polypeptides described herein. The other polypeptide can encode any peptidic or proteinaceous molecule, or a portion thereof, including, but not limited to an immunoglobulin, CD3, CD4, CD8, an MHC molecule, a CD1 molecule, e.g., CD1a, CD1b, CD1c, CD1d, etc. The fusion protein can comprise one or more copies of the inventive polypeptide and/or one or more copies of the other polypeptide. For instance, the fusion protein can comprise 1, 2, 3, 4, 5, or more, copies of the inventive polypeptide and/or of the other polypeptide. Suitable methods of making fusion proteins are known in the art, and include, for example, recombinant methods. In some aspects of the invention, the TCRs, polypeptides, and proteins of the invention may be expressed as a single protein comprising a linker peptide linking the α chain and the β chain. In this regard, the TCRs, polypeptides, and proteins of the invention may further comprise a linker peptide. The linker peptide may advantageously facilitate the expression of a recombinant TCR, polypeptide, and/or protein in a host cell. The linker peptide may comprise any suitable amino acid sequence. For example, the linker peptide may comprise the amino acid sequence of RAKRSGSGATNFSLLKQAGDVEENPGP (SEQ ID NO: 103). Upon expression of the construct including the linker peptide by a host cell, the linker peptide may be cleaved, resulting in separated α and β chains. In an aspect of the invention, the TCR, polypeptide, or protein may comprise an amino acid sequence comprising a full-length α chain, a full-length β chain, and a linker peptide positioned between the α and β chains. Leydig 770310 HHS E-054-2023-0-PC-01 23 In some aspects, the TCR, or protein disclosed herein comprises an α chain and/or a β chain, as disclosed herein, comprising a signal peptide. In some aspects, the sequence of the signal peptide of any of the α chains and/or β chains disclosed herein comprises an leucine, lysine, alanine or histidine residue substituted for the wild-type residue at position 2. In some aspects, the TCR, polypeptide or protein disclosed herein comprises a mature version of an α chain and/or a β chain, as disclosed herein, that lacks a signal peptide. The protein of the invention can be a recombinant antibody, or an antigen binding portion thereof, comprising at least one of the inventive polypeptides described herein. As used herein, “recombinant antibody” refers to a recombinant (e.g., genetically engineered) protein comprising at least one of the polypeptides of the invention and a polypeptide chain of an antibody, or an antigen binding portion thereof. The polypeptide of an antibody, or antigen binding portion thereof, can be a heavy chain, a light chain, a variable or constant region of a heavy or light chain, a single chain variable fragment (scFv), or an Fc, Fab, or F(ab)2' fragment of an antibody, etc. The polypeptide chain of an antibody, or an antigen binding portion thereof, can exist as a separate polypeptide of the recombinant antibody. Alternatively, the polypeptide chain of an antibody, or an antigen binding portion thereof, can exist as a polypeptide, which is expressed in frame (in tandem) with the polypeptide of the invention. The polypeptide of an antibody, or an antigen binding portion thereof, can be a polypeptide of any antibody or any antibody fragment, including any of the antibodies and antibody fragments described herein. The TCRs, polypeptides, and proteins of the invention can be of any length, i.e., can comprise any number of amino acids, provided that the TCRs, polypeptides, or proteins retain their biological activity, e.g., the ability to specifically bind to mutated p53; detect cancer in a mammal; or treat or prevent cancer in a mammal, etc. For example, the polypeptide can be in the range of from about 50 to about 5000 amino acids long, such as 50, 70, 75, 100, 125, 150, 175, 200, 300, 400, 500, 600, 700, 800, 900, 1000 or more amino acids in length. In this regard, the polypeptides of the invention also include oligopeptides. The TCRs, polypeptides, and proteins of the invention of the invention can comprise synthetic amino acids in place of one or more naturally-occurring amino acids. Such synthetic amino acids are known in the art, and include, for example, aminocyclohexane carboxylic acid, norleucine, α-amino n-decanoic acid, homoserine, S-acetylaminomethyl- cysteine, trans-3- and trans-4-hydroxyproline, 4-aminophenylalanine, 4-nitrophenylalanine, Leydig 770310 HHS E-054-2023-0-PC-01 24 4-chlorophenylalanine, 4- β-phenylserine β-hydroxyphenylalanine, phenylglycine, α-naphthylalanine, cyclohexylalanine, cyclohexylglycine, indoline-2- carboxylic acid, 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid, aminomalonic acid, aminomalonic acid monoamide, N’-benzyl-N’-methyl-lysine, N’,N’-dibenzyl-lysine, 6- hydroxylysine, ornithine, α-aminocyclopentane carboxylic acid, α-aminocyclohexane carboxylic acid, α-aminocycloheptane carboxylic acid, α-(2-amino-2-norbornane)-carboxylic acid, α,γ-diaminobutyric acid, α,β-diaminopropionic acid, homophenylalanine, and α-tert- butylglycine. The TCRs, polypeptides, and proteins of the invention can be, e.g., glycosylated, amidated, carboxylated, phosphorylated, esterified, N-acylated, cyclized via, e.g., a disulfide bridge, or converted into an acid addition salt and/or optionally dimerized or polymerized, or conjugated. The inventive TCRs, polypeptides, and proteins described herein (including any of the functional portions or variants thereof) are also contemplated to be useful as the soluble TCR component of bispecific engager TCR fusion proteins (e.g., IMMTAC (immune- mobilizing monoclonal TCRs against cancer) molecules). Bispecific engager TCR fusion proteins have two components. One component comprises a soluble TCR. The other component comprises an anti-CD3 effector. The anti-CD3 effector may be any molecule that engages with a CD3 molecule on T cells and activates a T cell immune response. For example, the anti-CD3 effector may be an anti-CD3 antibody or anti-CD3 antibody fragment. The soluble TCR component of the bispecific engager TCR fusion protein binds to the target antigen presented on the surface of cancer cells presented by an HLA molecule. The anti- CD3 effector component engages a CD3 molecule on T cells. The engagement of these components of the bispecific engager TCR fusion protein triggers the activation and recruitment of T cells and redirects T-cell killing to tumor cells. An aspect of the invention provides a bispecific engager TCR fusion protein comprising (i) any of the inventive TCRs, polypeptides, or proteins (including any of the functional portions or variants thereof) described herein and (ii) an anti-CD3 effector. Hereinafter, references to “protein(s)” also encompass the bispecific engager TCR fusion proteins described herein, unless specified otherwise. The TCR, polypeptide, and/or protein of the invention can be obtained by methods known in the art such as, for example, de novo synthesis. Also, polypeptides and proteins can be recombinantly produced using the nucleic acids described herein using standard Leydig 770310 HHS E-054-2023-0-PC-01 25 recombinant methods. See, for instance, and Sambrook, Molecular Cloning: A Laboratory Manual, 4th ed., Cold Spring Harbor Press, Cold Spring Harbor, NY (2012). Alternatively, the TCRs, polypeptides, and/or proteins described herein can be synthesized by any of a variety of commercial entities. In this respect, the inventive TCRs, polypeptides, and proteins can be synthetic, recombinant, isolated, and/or purified. An aspect of the invention provides a nucleic acid comprising a nucleotide sequence encoding any of the TCRs, polypeptides, or proteins described herein. “Nucleic acid,” as used herein, includes “polynucleotide,” “oligonucleotide,” and “nucleic acid molecule,” and generally means a polymer of DNA or RNA, which can be single-stranded or double-stranded, which can contain natural, non-natural or altered nucleotides, and which can contain a natural, non-natural or altered internucleotide linkage, such as a phosphoroamidate linkage or a phosphorothioate linkage, instead of the phosphodiester found between the nucleotides of an unmodified oligonucleotide. In an aspect, the nucleic acid comprises complementary DNA (cDNA). It is generally preferred that the nucleic acid does not comprise any insertions, deletions, inversions, and/or substitutions. However, it may be suitable in some instances, as discussed herein, for the nucleic acid to comprise one or more insertions, deletions, inversions, and/or substitutions. An aspect of the invention provides an isolated or purified nucleic acid comprising, from 5’ to 3’, a first nucleic acid sequence and a second nucleotide sequence, wherein the first and second nucleotide sequence, respectively, encode the amino sequences of SEQ ID NOs: 8 and 9; 9 and 8; 10 and 11; 11 and 10; 12 and 13; 13 and 12; 14 and 15; 15 and 14; 22 and 23; 23 and 22; 24 and 25; 25 and 24; 26 and 27; 27 and 26; 28 and 29; 29 and 28; 36 and 37; 37 and 36; 38 and 39; 39 and 38; 40 and 41; 41 and 40; 42 and 43; 43 and 42; 50 and 51; 51 and 50; 52 and 53; 53 and 52; 54 and 55; 55 and 54; 56 and 57; 57 and 56; 64 and 65; 65 and 64; 66 and 67; 67 and 66; 68 and 69; 69 and 68; 70 and 71; 71 and 70; 78 and 79; 79 and 78; 80 and 81; 81 and 80; 82 and 83; 83 and 82; 84 and 85; 85 and 84; 92 and 93; 93 and 92; 94 and 95; 95 and 94; 96 and 97; 97 and 96; 98 and 99; or 99 and 98. In an aspect of the invention, the nucleic acid further comprises a third nucleotide acid sequence interposed between the first and second nucleotide sequence, wherein the third nucleotide sequence encodes a cleavable linker peptide. For example, the cleavable linker peptide may comprise the amino acid sequence of RAKRSGSGATNFSLLKQAGDVEENPGP (SEQ ID NO: 103). Leydig 770310 HHS E-054-2023-0-PC-01 26 Preferably, the nucleic acids of invention are recombinant. As used herein, the term “recombinant” refers to (i) molecules that are constructed outside living cells by joining natural or synthetic nucleic acid segments to nucleic acid molecules that can replicate in a living cell, or (ii) molecules that result from the replication of those described in (i) above. For purposes herein, the replication can be in vitro replication or in vivo replication. The nucleic acids can be constructed based on chemical synthesis and/or enzymatic ligation reactions using procedures known in the art. See, for example, Green and Sambrook et al., supra. For example, a nucleic acid can be chemically synthesized using naturally occurring nucleotides or variously modified nucleotides designed to increase the biological stability of the molecules or to increase the physical stability of the duplex formed upon hybridization (e.g., phosphorothioate derivatives and acridine substituted nucleotides). Examples of modified nucleotides that can be used to generate the nucleic acids include, but are not limited to, 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xanthine, 4-acetylcytosine, 5-(carboxyhydroxymethyl) uracil, 5-carboxymethylaminomethyl- 2-thiouridine, 5-carboxymethylaminomethyluracil, dihydrouracil, beta-D-galactosylqueosine, inosine, N6-isopentenyladenine, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2- methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-substituted adenine, 7-methylguanine, 5-methylaminomethyluracil, 5-methoxyaminomethyl-2-thiouracil, beta-D-mannosylqueosine, 5'-methoxycarboxymethyluracil, 5-methoxyuracil, 2-methylthio- N6-isopentenyladenine, uracil-5-oxyacetic acid (v), wybutoxosine, pseudouracil, queosine, 2- thiocytosine, 5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, uracil-5- oxyacetic acid methylester, 3-(3-amino-3-N-2-carboxypropyl) uracil, and 2,6-diaminopurine. Alternatively, one or more of the nucleic acids of the invention can be synthesized by any of a variety of commercial entities. In an aspect of the invention, the nucleic acid comprises a codon-optimized nucleotide sequence encoding any of the TCRs, polypeptides, or proteins described herein. Without being bound to any particular theory or mechanism, it is believed that codon optimization of the nucleotide sequence increases the translation efficiency of the mRNA transcripts. Codon optimization of the nucleotide sequence may involve substituting a native codon for another codon that encodes the same amino acid, but can be translated by tRNA that is more readily available within a cell, thus increasing translation efficiency. Optimization of the nucleotide sequence may also reduce secondary mRNA structures that would interfere with translation, thus increasing translation efficiency. Leydig 770310 HHS E-054-2023-0-PC-01 27 An aspect of the invention also a nucleic acid comprising a nucleotide sequence which is complementary to the nucleotide sequence of any of the nucleic acids described herein. The nucleic acids of the invention can be incorporated into a recombinant expression vector. In this regard, an aspect of the invention provides a recombinant expression vector comprising any of the nucleic acids of the invention. In an aspect of the invention, the recombinant expression vector comprises a nucleotide sequence encoding the α chain, the β chain, and linker peptide. For purposes herein, the term “recombinant expression vector” means a genetically-modified oligonucleotide or polynucleotide construct that permits the expression of an mRNA, protein, polypeptide, or peptide by a host cell, when the construct comprises a nucleotide sequence encoding the mRNA, protein, polypeptide, or peptide, and the vector is contacted with the cell under conditions sufficient to have the mRNA, protein, polypeptide, or peptide expressed within the cell. The vectors of the invention are not naturally-occurring as a whole. However, parts of the vectors can be naturally-occurring. The inventive recombinant expression vectors can comprise any type of nucleotide, including, but not limited to DNA and RNA, which can be single-stranded or double-stranded, synthesized or obtained in part from natural sources, and which can contain natural, non-natural or altered nucleotides. The recombinant expression vectors can comprise naturally-occurring, non- naturally-occurring internucleotide linkages, or both types of linkages. Preferably, the non- naturally occurring or altered nucleotides or internucleotide linkages do not hinder the transcription or replication of the vector. The recombinant expression vector of the invention can be any suitable recombinant expression vector, and can be used to transform or transfect any suitable host cell. Suitable vectors include those designed for propagation and expansion or for expression or both, such as plasmids and viruses. The vector can be selected from the group consisting of the transposon/transposase series, pUC series (Fermentas Life Sciences), the pBluescript series (Stratagene, LaJolla, CA), the pET series (Novagen, Madison, WI), the pGEX series (Pharmacia Biotech, Uppsala, Sweden), and the pEX series (Clontech, Palo Alto, CA). Bacteriophage vectors, such as λGT10, λGT11, λZapII (Stratagene), λEMBL4, and λNM1149, also can be used. Examples of animal expression vectors include pEUK-Cl, pMAM and pMAMneo (Clontech). Preferably, the recombinant expression vector is a transposon or a viral vector, e.g., a lentiviral vector or a retroviral vector. Leydig 770310 HHS E-054-2023-0-PC-01 28 The recombinant expression of the invention can be prepared using standard recombinant DNA techniques described in, for example, Green and Sambrook et al., supra. Constructs of expression vectors, which are circular or linear, can be prepared to contain a replication system functional in a prokaryotic or eukaryotic host cell. Replication systems can be derived, e.g., from ColEl, 2 μ plasmid, λ, SV40, bovine papillomavirus, and the like. Desirably, the recombinant expression vector comprises regulatory sequences, such as transcription and translation initiation and termination codons, which are specific to the type of host cell (e.g., bacterium, fungus, plant, or animal) into which the vector is to be introduced, as appropriate and taking into consideration whether the vector is DNA- or RNA- based. The recombinant expression vector can include one or more marker genes, which allow for selection of transformed or transfected host cells. Marker genes include biocide resistance, e.g., resistance to antibiotics, heavy metals, etc., complementation in an auxotrophic host cell to provide prototrophy, and the like. Suitable marker genes for the inventive expression vectors include, for instance, neomycin/G418 resistance genes, hygromycin resistance genes, histidinol resistance genes, tetracycline resistance genes, and ampicillin resistance genes. The recombinant expression vector can comprise a native or nonnative promoter operably linked to the nucleotide sequence encoding the TCR, polypeptide, or protein, or to the nucleotide sequence which is complementary to the nucleotide sequence encoding the TCR, polypeptide, or protein. The selection of promoters, e.g., strong, weak, inducible, tissue-specific and developmental-specific, is within the ordinary skill of the artisan. Similarly, the combining of a nucleotide sequence with a promoter is also within the skill of the artisan. The promoter can be a non-viral promoter, e.g., a human elongation factor-1α promoter, or a viral promoter, e.g., a cytomegalovirus (CMV) promoter, an SV40 promoter, an RSV promoter, and a promoter found in the long-terminal repeat of the murine stem cell virus. The inventive recombinant expression vectors can be designed for either transient expression, for stable expression, or for both. Also, the recombinant expression vectors can be made for constitutive expression or for inducible expression. Further, the recombinant expression vectors can be made to include a suicide gene. As used herein, the term “suicide gene” refers to a gene that causes the cell expressing Leydig 770310 HHS E-054-2023-0-PC-01 29 the suicide gene to die. The suicide gene can a gene that confers sensitivity to an agent, e.g., a drug, upon the cell in which the gene is expressed, and causes the cell to die when the cell is contacted with or exposed to the agent. Suicide genes are known in the art and include, for example, the Herpes Simplex Virus (HSV) thymidine kinase (TK) gene, cytosine daminase, purine nucleoside phosphorylase, and nitroreductase. Another aspect of the invention provides an isolated or purified TCR, polypeptide, or protein encoded by any of the nucleic acids or vectors described herein with respect to other aspects of the invention. Still another aspect of the invention provides an isolated or purified TCR, polypeptide, or protein that results from expression of any of the nucleic acids or vectors, described herein with respect to other aspects of the invention, in a cell. Another aspect of the invention further provides a host cell comprising any of the nucleic acids or any of the recombinant expression vectors described herein. As used herein, the term “host cell” refers to any type of cell that can contain the inventive recombinant expression vector. The host cell can be a eukaryotic cell, e.g., plant, animal, fungi, or algae, or can be a prokaryotic cell, e.g., bacteria or protozoa. The host cell can be a cultured cell or a primary cell, i.e., isolated directly from an organism, e.g., a human. The host cell can be an adherent cell or a suspended cell, i.e., a cell that grows in suspension. Suitable host cells are known in the art and include, for instance, DH5α E. coli cells, Chinese hamster ovarian cells, monkey VERO cells, COS cells, HEK293 cells, and the like. For purposes of amplifying or replicating the recombinant expression vector, the host cell is preferably a prokaryotic cell, e.g., a DH5α cell. For purposes of producing a recombinant TCR, polypeptide, or protein, the host cell is preferably a mammalian cell. Most preferably, the host cell is a human cell. For example, the host cell may be a human lymphocyte. In an aspect of the invention, the host cell is selected from the group consisting of a T cell, a natural killer T (NKT) cell, an invariant natural killer T (iNKT) cell, a natural killer (NK) cell, a macrophage, a pluripotent cell, and a multipotent cell. While the host cell can be of any cell type, can originate from any type of tissue, and can be of any developmental stage, the host cell preferably is a peripheral blood lymphocyte (PBL) or a peripheral blood mononuclear cell (PBMC). More preferably, the host cell is a T cell. For purposes herein, the T cell can be any T cell, such as a cultured T cell, e.g., a primary T cell, or a T cell from a cultured T cell line, e.g., Jurkat, SupT1, etc., or a T cell obtained from a mammal. If obtained from a mammal, the T cell can be obtained from Leydig 770310 HHS E-054-2023-0-PC-01 30 numerous sources, including but not limited blood, bone marrow, lymph node, the thymus, or other tissues or fluids. T cells can also be enriched for or purified. Preferably, the T cell is a human T cell. The T cell can be any type of T cell and can be of any developmental stage, including but not limited to, CD4+/CD8+ double positive T cells, CD4+ helper T cells, e.g., Th1 and Th2 cells, CD4+ T cells, CD8+ T cells (e.g., cytotoxic T cells), tumor infiltrating lymphocytes (TILs), memory T cells (e.g., central memory T cells and effector memory T cells), naïve T cells, and the like. In an aspect of the invention, the host cell is a pluripotent cell or a multipotent cell. Pluripotent cells have the capacity to give rise to any of the three germ layers: endoderm, mesoderm, and ectoderm. Pluripotent cells may comprise, for example, stem cells, e.g., embryonic stem cells, nuclear transfer derived embryonic stem cells, induced pluripotent stem cells (iPSC), etc. Multipotent cells may comprise, for example, hematopoietic stem cells. Modifying, e.g., reprogramming, cells to a pluripotent state refers to the reversion of a cell to a pluripotent cell and is described for example, in Crompton et al., Trends Immunol., 35(4): 178-185 (2014). Exemplary techniques may include somatic cell nuclear transfer (SCNT), cell–cell fusion, and direct reprogramming. Examples of methods for carrying out cell-cell fusion are described, for example, in Ogle et al., Nat. Rev. Mol. Cell Biol.6: 567-75 (2005) and Zhou et al., Cell Stem Cell, 3: 382-388 (2008). Examples of methods for carrying out SCNT are described, for example, in Hanna et al., Cell, 143: 508- 525 (2010); Stadtfeld et al., Genes Dev., 24: 2239-2263 (2010); Wilmut et al., Nature, 385: 810-813 (1997); Vizcardo et al., Cell Stem Cell, 12: 31-36 (2013); and Crompton et al., Cell Stem Cell, 12: 6-8 (2013). In an aspect of the invention, the host cell is an iPSC that was prepared by reprogramming, any of the host cells described herein (e.g., T cells, NK cells, or invariant natural killer T cells) to a pluripotent state. Also provided by an aspect of the invention is a population of cells comprising at least one host cell described herein. The population of cells can be a heterogeneous population comprising the host cell comprising any of the recombinant expression vectors described, in addition to at least one other cell, e.g., a host cell (e.g., a T cell), which does not comprise any of the recombinant expression vectors, or a cell other than a T cell, e.g., a B cell, a macrophage, a neutrophil, an erythrocyte, a hepatocyte, an endothelial cell, an epithelial cell, a muscle cell, a brain cell, etc. Alternatively, the population of cells can be a substantially homogeneous population, in which the population comprises mainly of host cells (e.g., consisting essentially of) comprising the recombinant expression vector. The Leydig 770310 HHS E-054-2023-0-PC-01 31 population also can be a clonal population in which all cells of the population are clones of a single host cell comprising a recombinant expression vector, such that all cells of the population comprise the recombinant expression vector. In one aspect of the invention, the population of cells is a clonal population comprising host cells comprising a recombinant expression vector as described herein. In an aspect of the invention, the numbers of cells in the population may be rapidly expanded. Expansion of the numbers of T cells can be accomplished by any of a number of methods as are known in the art as described in, for example, U.S. Patent Nos. 8,034,334; 8,383,099; and 11,401,503; Dudley et al., J. Immunother., 26:332-42 (2003); and Riddell et al., J. Immunol. Methods, 128:189-201 (1990). In an aspect, expansion of the numbers of T cells is carried out by culturing the T cells with OKT3 antibody, IL-2, and feeder PBMC (e.g., irradiated allogeneic PBMC). An aspect of the invention provides a method of producing any of the TCRs, polypeptides, or proteins described herein, the method comprising culturing any of the host cells or populations of host cells described herein, so that the TCR, polypeptide, or protein is produced. Another aspect of the invention is a method of producing a host cell expressing a TCR that has antigenic specificity for a human p53Y220C or human p53R175H amino acid sequence, the method comprising contacting a cell with any of the inventive recombinant expression vectors described herein under conditions that allow introduction of the vector into the cell. Another aspect of the invention provides a method of producing an engineered human cell (or an engineered population of human cells), the method comprising introducing any of the inventive nucleic acids or recombinant expression vectors described herein to an isolated human cell (or an isolated population of human cells), wherein the nucleic acid or recombinant expression vector comprises a nucleotide sequence encoding any of the inventive TCRs, polypeptides, or proteins described herein. The isolated human cell, or isolated population of human cells, to which the nucleic acid or recombinant expression vector is introduced, may be as described herein with respect to other aspects of the invention. The inventive TCRs, polypeptides, proteins, nucleic acids, recombinant expression vectors, and host cells (including populations thereof), can be isolated and/or purified. The term “isolated” as used herein means having been removed from its natural Leydig 770310 HHS E-054-2023-0-PC-01 32 environment. The term “purified” as used means having been increased in purity, wherein “purity” is a relative term, and not to be necessarily construed as absolute purity. For example, the purity can be at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or can be about 100%. The inventive TCRs, polypeptides, proteins, nucleic acids, recombinant expression vectors, and host cells (including populations thereof), all of which are collectively referred to as “inventive TCR materials” hereinafter, can be formulated into a composition, such as a pharmaceutical composition. In this regard, an aspect of the invention provides a pharmaceutical composition comprising any of the TCRs, polypeptides, proteins, nucleic acids, expression vectors, and host cells (including populations thereof), described herein, and a pharmaceutically acceptable carrier. The inventive pharmaceutical compositions containing any of the inventive TCR materials can comprise more than one inventive TCR material, e.g., a polypeptide and a nucleic acid, or two or more different TCRs. Alternatively, the pharmaceutical composition can comprise an inventive TCR material in combination with another pharmaceutically active agent(s) or drug(s), such as a chemotherapeutic agent, e.g., asparaginase, busulfan, carboplatin, cisplatin, daunorubicin, doxorubicin, fluorouracil, gemcitabine, hydroxyurea, methotrexate, paclitaxel, rituximab, vinblastine, vincristine, etc. Preferably, the carrier is a pharmaceutically acceptable carrier. With respect to pharmaceutical compositions, the carrier can be any of those conventionally used for the particular inventive TCR material under consideration. Methods for preparing administrable compositions are known or apparent to those skilled in the art and are described in more detail in, for example, Remington: The Science and Practice of Pharmacy, 23rd Ed., Academic Press (2020). It is preferred that the pharmaceutically acceptable carrier be one which has no detrimental side effects or toxicity under the conditions of use. The choice of carrier will be determined in part by the particular inventive TCR material, as well as by the particular method used to administer the inventive TCR material. Accordingly, there are a variety of suitable formulations of the pharmaceutical composition of the invention. Suitable formulations may include any of those for parenteral, subcutaneous, intravenous, intramuscular, intraarterial, intrathecal, intratumoral, or interperitoneal administration. More than one route can be used to administer the inventive TCR materials, and in certain instances, a particular route can provide a more immediate and more effective response than another route. Leydig 770310 HHS E-054-2023-0-PC-01 33 Preferably, the inventive TCR is administered by injection, e.g., intravenously. When the inventive TCR material is a host cell expressing the inventive TCR, the pharmaceutically acceptable carrier for the cells for injection may include any isotonic carrier such as, for example, normal saline (about 0.90% w/v of NaCl in water, about 300 mOsm/L NaCl in water, or about 9.0 g NaCl per liter of water), NORMOSOL R electrolyte solution (Abbott, Chicago, IL), PLASMA-LYTE A (Baxter, Deerfield, IL), about 5% dextrose in water, or Ringer's lactate. In an aspect, the pharmaceutically acceptable carrier is supplemented with human serum albumin. The amount or dose (e.g., numbers of cells when the inventive TCR material is one or more cells) of the inventive TCR material administered should be sufficient to effect, e.g., a therapeutic or prophylactic response, in the subject or animal over a reasonable time frame. For example, the dose of the inventive TCR material should be sufficient to bind to a cancer antigen (e.g., mutated p53), or detect, treat or prevent cancer in a period of from about 2 hours or longer, e.g., 12 to 24 or more hours, from the time of administration. In certain aspects, the time period could be even longer. The dose will be determined by the efficacy of the particular inventive TCR material and the condition of the animal (e.g., human), as well as the body weight of the animal (e.g., human) to be treated. Many assays for determining an administered dose are known in the art. For example, an assay, which comprises comparing the extent to which target cells are lysed or IFN-γ is secreted by T cells expressing the inventive TCR, polypeptide, or protein upon administration of a given dose of such T cells to a mammal among a set of mammals of which each is given a different dose of the T cells, could be used to determine a starting dose to be administered to a mammal. The extent to which target cells are lysed or IFN-γ is secreted upon administration of a certain dose can be assayed by methods known in the art. The dose of the inventive TCR material also will be determined by the existence, nature and extent of any adverse side effects that might accompany the administration of a particular inventive TCR material. Typically, the attending physician will decide the dosage of the inventive TCR material with which to treat each individual patient, taking into consideration a variety of factors, such as age, body weight, general health, diet, sex, inventive TCR material to be administered, route of administration, and the severity of the cancer being treated. In an aspect in which the inventive TCR material is a population of cells, the number of cells administered per infusion may vary, e.g., from about 1 x 106 to about 1 x 1012 cells or more. In certain aspects, fewer than 1 x 106 cells may be administered. Leydig 770310 HHS E-054-2023-0-PC-01 34 One of ordinary skill in the art readily appreciate that the inventive TCR materials of the invention can be modified in any number of ways, such that the therapeutic or prophylactic efficacy of the inventive TCR materials is increased through the modification. For instance, the inventive TCR materials can be conjugated either directly or indirectly through a bridge to a chemotherapeutic agent. The practice of conjugating compounds to a chemotherapeutic agent is known in the art. One of ordinary skill in the art recognizes that sites on the inventive TCR materials, which are not necessary for the function of the inventive TCR materials, are ideal sites for attaching a bridge and/or a chemotherapeutic agent, provided that the bridge and/or chemotherapeutic agent, once attached to the inventive TCR materials, do(es) not interfere with the function of the inventive TCR materials, i.e., the ability to bind to mutated p53 or to detect, treat, or prevent cancer. It is contemplated that the inventive pharmaceutical compositions, TCRs, polypeptides, proteins, nucleic acids, recombinant expression vectors, host cells, or populations of cells can be used in methods of treating or preventing cancer. Without being bound to a particular theory, the inventive TCRs are believed to bind specifically to mutated p53, such that the TCR (or related inventive polypeptide or protein), when expressed by a cell, is able to mediate an immune response against a target cell expressing mutated p53. In this regard, an aspect of the invention provides a method of treating or preventing cancer in a mammal, comprising administering to the mammal any of the pharmaceutical compositions, TCRs, polypeptides, or proteins described herein, any nucleic acid or recombinant expression vector comprising a nucleotide sequence encoding any of the TCRs, polypeptides, proteins described herein, or any host cell or population of cells comprising a recombinant vector which encodes any of the TCRs, polypeptides, or proteins described herein, in an amount effective to treat or prevent cancer in the mammal. An aspect of the invention provides any of the pharmaceutical compositions, TCRs, polypeptides, or proteins described herein, any nucleic acid or recombinant expression vector comprising a nucleotide sequence encoding any of the TCRs, polypeptides, proteins described herein, or any host cell or population of cells comprising a recombinant vector which encodes any of the TCRs, polypeptides, or proteins described herein, for use in the treatment or prevention of cancer in a mammal. The terms “treat,” and “prevent” as well as words stemming therefrom, as used herein, do not necessarily imply 100% or complete treatment or prevention. Rather, there are varying degrees of treatment or prevention of which one of ordinary skill in the art recognizes Leydig 770310 HHS E-054-2023-0-PC-01 35 as having a potential benefit or therapeutic In this respect, the inventive methods can provide any amount of any level of treatment or prevention of cancer in a mammal. Furthermore, the treatment or prevention provided by the inventive method can include treatment or prevention of one or more conditions or symptoms of the cancer being treated or prevented. For example, treatment or prevention can include promoting the regression of a tumor. Also, for purposes herein, “prevention” can encompass delaying the onset of the cancer, or a symptom or condition thereof. Alternatively or additionally, “prevention” may encompass preventing or delaying the recurrence of cancer, or a symptom or condition thereof. It is also contemplated that the inventive pharmaceutical compositions, TCRs, polypeptides, proteins, nucleic acids, recombinant expression vectors, host cells, or populations of cells can be used in methods of inducing an immune response against a cancer in a mammal. In this regard, an aspect of the invention provides a method of inducing an immune response against a cancer in a mammal, comprising administering to the mammal any of the pharmaceutical compositions, TCRs, polypeptides, or proteins described herein, any nucleic acid or recombinant expression vector comprising a nucleotide sequence encoding any of the TCRs, polypeptides, proteins described herein, or any host cell or population of cells comprising a recombinant vector which encodes any of the TCRs, polypeptides, or proteins described herein, in an amount effective to induce an immune response against the cancer in the mammal. An aspect of the invention provides any of the pharmaceutical compositions, TCRs, polypeptides, or proteins described herein, any nucleic acid or recombinant expression vector comprising a nucleotide sequence encoding any of the TCRs, polypeptides, proteins described herein, or any host cell or population of cells comprising a recombinant vector which encodes any of the TCRs, polypeptides, or proteins described herein, for use in the inducement of an immune response against a cancer in a mammal. Also provided by an aspect of the invention is a method of detecting the presence of cancer in a mammal. The method comprises (i) contacting a sample comprising one or more cells from the mammal with any of the inventive TCRs, polypeptides, proteins, nucleic acids, recombinant expression vectors, host cells, populations of cells, or pharmaceutical compositions described herein, thereby forming a complex, and (ii) detecting the complex, wherein detection of the complex is indicative of the presence of cancer in the mammal. Leydig 770310 HHS E-054-2023-0-PC-01 36 With respect to the inventive of detecting cancer in a mammal, the sample of cells can be a sample comprising whole cells, lysates thereof, or a fraction of the whole cell lysates, e.g., a nuclear or cytoplasmic fraction, a whole protein fraction, or a nucleic acid fraction. For purposes of the inventive detecting method, the contacting can take place in vitro or in vivo with respect to the mammal. Preferably, the contacting is in vitro. Also, detection of the complex can occur through any number of ways known in the art. For instance, the inventive TCRs, polypeptides, proteins, nucleic acids, recombinant expression vectors, host cells, or populations of cells, described herein, can be labeled with a detectable label such as, for instance, a radioisotope, a fluorophore (e.g., fluorescein isothiocyanate (FITC), phycoerythrin (PE)), an enzyme (e.g., alkaline phosphatase, horseradish peroxidase), and element particles (e.g., gold particles). For purposes of the inventive methods, wherein host cells or populations of cells are administered, the cells can be cells that are allogeneic or autologous to the mammal. Preferably, the cells are autologous to the mammal. With respect to the inventive methods, the cancer can be any cancer, including, e.g., any of acute lymphocytic cancer, acute myeloid leukemia, alveolar rhabdomyosarcoma, bone cancer, brain cancer, breast cancer, cancer of the anus, anal canal, or anorectum, cancer of the eye, cancer of the intrahepatic bile duct, cancer of the joints, cancer of the neck, gallbladder, or pleura, cancer of the nose, nasal cavity, or middle ear, cancer of the oral cavity, cancer of the vagina, cancer of the vulva, chronic lymphocytic leukemia, chronic myeloid cancer, colon cancer, colocrectal cancer, endometrial cancer, esophageal cancer, uterine cervical cancer, gastrointestinal carcinoid tumor, glioma, Hodgkin lymphoma, hypopharynx cancer, kidney cancer, larynx cancer, liver cancer, lung cancer, malignant mesothelioma, melanoma, multiple myeloma, nasopharynx cancer, non-Hodgkin lymphoma, cancer of the oropharynx, ovarian cancer, cancer of the penis, pancreatic cancer, peritoneum, omentum, and mesentery cancer, pharynx cancer, prostate cancer, rectal cancer, renal cancer, skin cancer, small intestine cancer, soft tissue cancer, stomach cancer, testicular cancer, thyroid cancer, cancer of the uterus, ureter cancer, and urinary bladder cancer. In a preferred aspect, the cancer is a cancer which expresses mutated p53. The cancer may express p53 with a mutation at one or both of positions 220 and 175, as defined by SEQ ID NO: 1. The cancer may express p53 with one or both of the following human p53 mutations: Y220C and R175H. In an aspect of the invention, the cancer is an epithelial cancer. In an aspect of the Leydig 770310 HHS E-054-2023-0-PC-01 37 invention, the cancer is colon cancer, rectal cancer, ovarian cancer, endometrial cancer, non-small cell lung cancer (NSCLC), glioblastoma, uterine cervical cancer, head and neck cancer, breast cancer, pancreatic cancer, or bladder cancer. The cancer may be known to comprise a Y220C or R175H mutation in human p53. The mammal referred to in the inventive methods can be any mammal. As used herein, the term “mammal” refers to any mammal, including, but not limited to, mammals of the order Rodentia, such as mice and hamsters, and mammals of the order Lagomorpha, such as rabbits. It is preferred that the mammals are from the order Carnivora, including Felines (cats) and Canines (dogs). It is more preferred that the mammals are from the order Artiodactyla, including Bovines (cows) and Swines (pigs) or of the order Perssodactyla, including Equines (horses). It is most preferred that the mammals are of the order Primates, Ceboids, or Simoids (monkeys) or of the order Anthropoids (humans and apes). An especially preferred mammal is the human. The following examples further illustrate the invention but, of course, should not be construed as in any way limiting its scope. EXAMPLE 1 This example demonstrates the isolation of TCRs from Patient 4196. The p53R175H 9-amino acid epitope (HMTEVVRHC) (SEQ ID NO: 112) contains cysteine at its C-terminus. Cysteine residues can oxidize and form disulfide bonds with other cysteine residues under oxidizing conditions, thus potentially interfering with their ability to bind to MHC molecules (Sachs et al., J. Immunol., 205(2): 539–549 (2020)). This may lead to reduced T cell activity. To minimize that effect, in the experiments described in the Examples below, the native cysteine at position 9 of (HMTEVVRHC) (SEQ ID NO: 112) was replaced with alpha-aminobutyric acid (AABA) to produce the p53R175H analogue peptide HMTEVVRHX (SEQ ID NO: 116), wherein X at position 9 of SEQ ID NO: 116 is AABA. AABA is a cysteine analogue containing a methyl group in place of the sulfhydryl group present in cysteine. The native cysteine at position 9 of (HMTEVVRRC) (SEQ ID NO: 113) was replaced with AABA to produce the corresponding wild-type analogue peptide HMTEVVRRX (SEQ ID NO: 117), wherein X at position 9 of SEQ ID NO: 117 is AABA. Three different culture conditions were tested prior to isolating TCRs specific for p53R175H from TILs from Patient 4196 (4196 TILs). Leydig 770310 HHS E-054-2023-0-PC-01 38 Tetramer staining of 4196 TILs was performed for the detection of p53R175H- reactive cells. Following two weeks of the cultures outlined in Table 1, T cells were stained for the HLA-A2 tetramer. The HLA-A2 tetramer was oligomerized HLA-A2 monomers loaded with the p53R175H 9-amino acid epitope analogue HMTEVVRHX (SEQ ID NO: 116), wherein X at position 9 of SEQ ID NO: 116 is AABA. The tetramer positive cell frequencies observed for each of culture conditions 1, 2, and 3 are shown in Figure 1A. TABLE 1 Culture condition 1 Antigen (p53R175H)-specific expansion condition Culture condition 2 Culture with anti-CD3 antibod feeder cells and IL-2
Figure imgf000040_0001
showed specific recognition of antigen presenting cells (APCs) pulsed with p53R175H peptide analogue but not the corresponding wild-type (WT) peptide analogue. Subsequently, fluorescence-activated cell sorting (FACS) was performed based on upregulation of the T cell activation markers, 4-1BB and OX-40. The three TCRs were the 4196-C TCR (TRAV38- 1/TRBV5-1), 4196-D TCR (TRAV14/ TRBV7-9), and 4196-E TCR (TRAV10/TRBV6-6). The sequences of the TCR alpha and beta chain variable regions were identified by single-cell TCR sequencing. The amino acid sequences of the alpha and beta chain variable regions are shown in Table 2. The CDRs are underlined. The N-terminal signal peptides are in bold font. TABLE 2 TCR Name TCR chain Amino acid sequence Variable α QTVTQSQPEMSVQEAETVTLSCTYDTSENNYYLFWYKQPPSR D F L T N G
Figure imgf000040_0002
Leydig 770310 HHS E-054-2023-0-PC-01 39 TCR Name TCR chain Amino sequence Variable β MGSRLLCWVLLCLLGAGPVKAGVTQTPRYLIKTRGQQVTLS S G G D G D Y K F R G G S L S L T C P
Figure imgf000041_0001
This example demonstrates the isolation of a TCR from Patient 4424. TILs from Patient 4424 were subjected to tetramer staining for the detection of p53R175H-reactive cells. Following two weeks of the cultures outlined in Table 1, T cells were Leydig 770310 HHS E-054-2023-0-PC-01 40 stained for the HLA-A2 tetramer. The tetramer was oligomerized HLA-A2 loaded with the p53R175H 9-amino acid epitope analogue HMTEVVRHX (SEQ ID NO: 116), wherein X at position 9 of SEQ ID NO: 116 is AABA. The flow cytometry staining for CD8 and tetramer is shown in Figure 2A. The 4424-R175H TCR was isolated from the tetramer. Because the tetramer was built of HLA-A2 monomers, these staining data suggest that the 4424-R175H TCR would be restricted by HLA-A2. While the other TCRs targeting mutant p53 were previously identified by co-culture experiments, the 4424-R175H TCR (TRAV10/TRBV2) was the first TCR that was identified by use of a tetramer. The sequences of the TCR alpha and beta chain variable regions were identified by single-cell TCR sequencing. The amino acid sequences of the alpha and beta chain variable regions are shown in Table 3. The CDRs are underlined. The N-terminal signal peptides are in bold font. TABLE 3 TCR Name TCR chain Amino acid sequence Variable α KNQVEQSPQSLIILEGKNCTLQCNYTVSPFSNLRWYKQDTGRG S E M L A R S T
Figure imgf000042_0001
This example demonstrates the isolation of TCRs from Patient 4402. TIL from Patient 4402 showed specific recognition of antigen presenting cells (APCs) pulsed with p53Y220C peptide (DRNTFRHSVVVPCEPPEVGSDCTTI) (SEQ ID NO: 114), but not the corresponding WT peptide (DRNTFRHSVVVPYEPPEVGSDCTTI) (SEQ Leydig 770310 HHS E-054-2023-0-PC-01 41 ID NO: 115). Subsequntly, FACS was based on upregulation of the T cell activation markers, 4-1BB and OX-40. Twenty-one (21) TCRs (4402-Y220C-A to -U, respectively) were isolated from TIL from Patient 4402. ELISpot analysis was performed to detect IFN-γ release following co-culture of healthy donor PBL expressing the respective 21 TCRs that were isolated from Patient 4402 TIL (effector cells). The 4402-Y220C-B TCR, 4402-Y220C-C TCR, and 4402-Y220C-L TCR showed increased IFN-γ release against B cells pulsed with 1 µg/mL mutant p53 Y220C peptide (DRNTFRHSVVVPCEPPEVGSDCTTI) (SEQ ID NO: 114) (target cells) relative to the DMSO controls, while the other 18 TCRs did not. Effector cells treated with phorbol 12- myristate 13-acetate (PMA) and ionomycin were used as a positive control. Controls included: cultures with only effector cells, cultures with target cells pulsed with DMSO, and cultures with PBL transduced with an empty vector (mock). The sequences of the TCR alpha and beta chain variable regions were identified by single-cell TCR sequencing. The amino acid sequences of the alpha and beta chain variable regions of three of the TCRs (4402-Y220C-B TCR (TRAV12-3/TRBV7-2), 4402- Y220C-C TCR (TRAV26-2/TRBV5-6), and 4402-Y220C-L TCR (TRAV23/TRBV5-1)) are shown in Table 4. The CDRs are underlined. The N-terminal signal peptides are in bold font. TABLE 4 TCR Name TCR chain Amino acid sequence Variable α QQKEVEQDPGPLSVPEGAIVSLNCTYSNSAFQYFMWYRQYSR A E A I F L R E
Figure imgf000043_0001
Leydig 770310 HHS E-054-2023-0-PC-01 42 TCR Name TCR chain Amino sequence Variable α KTTQPNSMESNEEEPVHLPCNHSTISGTDYIHWYRQLPSQGPE YVIHGLTSNVNNRMASLAIAEDRKSSTLILHRATLRDAAVYYC Q A S R K F G Y O: F L IV K N S S
Figure imgf000044_0001
This example demonstrates the construction of retroviral vectors encoding the respective TCRs of Examples 1-3. Nucleotide sequences encoding the variable regions of the α and β chains of the TCRs of Tables 2-4 were obtained and codon optimized. The TCRβ VDJ regions were fused to the mouse TCRβ constant chain. The TCRα VJ regions were fused to the mouse TCRα constant chain. Without being bound to a particular theory or mechanism, it is believed that Leydig 770310 HHS E-054-2023-0-PC-01 43 replacing the constant regions of the human and TCRβ chains with the corresponding murine constant regions improves TCR expression and functionality (Cohen et al., Cancer Res., 66(17): 8878-86 (2006)). In addition, the murine TCRα and TCRβ constant chains were cysteine-modified. Transmembrane hydrophobic mutations were introduced into the murine TCRα constant chain. Without being bound to a particular theory or mechanism, it is believed that these modifications result in preferential pairing of the introduced TCR chains and enhanced TCR surface expression and functionality (Cohen et al., Cancer Res., 67(8):3898-903 (2007); Haga-Friedman et al., J. Immu., 188: 5538–5546 (2012)). The full length α and β chains of each of the seven TCRs, including these modifications to the constant region, are shown in Table 5. In Table 5, the CDRs are underlined, the constant region is in italics, and the modified amino acid residues of the constant region are underlined and in bold. TABLE 5 TCR TCR chain Amino acid sequence Name D K A N P D S L VI Y D E M F G R H
Figure imgf000045_0001
Leydig 770310 HHS E-054-2023-0-PC-01 44 TCR TCR chain Amino acid sequence Name NPRNHFRCQVQFHGLSEEDKWPEGSPKPVTQNISAEAWGRADCGITSASY ) T A K Q E T K P P V I G Q N L SP L F S : S Q G L F P D S L T V Y
Figure imgf000046_0001
Leydig 770310 HHS E-054-2023-0-PC-01 45 TCR TCR chain Amino acid sequence Name PSSDVPCDATLTEKSFETDMNLNFQNLLVIVLRILLLKVAGFNLLMTLRLW Y S L F S : S P S F L S K V T T T F A H Y ) Y Y A K Q G T K P P
Figure imgf000047_0001
Leydig 770310 HHS E-054-2023-0-PC-01 46 TCR TCR chain Amino acid sequence Name KPVTQNISAEAWGRADCGITSASYQQGVLSATILYEILLGKATLYAVLVSTLV L I S T T Q A A W A T P W L G E K P P V P P S S R H Y ) S D
Figure imgf000048_0001
Leydig 770310 HHS E-054-2023-0-PC-01 47 TCR TCR chain Amino acid sequence Name MKAMDSKSNGAIAWSNQTSFTCQDIFKETNATYPSSDVPCDATLTEKSFET P D S L G C F D N F G H Y )
Figure imgf000049_0001
, q g re cloned into a MSGV1-based retroviral vector. The TCRβ and TCRα chains were separated by a Furin Ser/Gly P2A linker RAKRSGSGATNFSLLKQAGDVEENPGP (SEQ ID NO: 103). Without being bound to a particular theory or mechanism, it is believed that the linker provides comparable expression efficiency of the two chains (Szymczak et al., Nat. Biotechnol., 22(5):589-94 (2004)). EXAMPLE 5 This example demonstrates the specificity and avidity of the 4196-C TCR, 4196-D TCR, and 4196-E TCR. Healthy donor PBL were transduced with the retroviral vector encoding the 4196- C TCR, 4196-D TCR, or 4196-E TCR of Example 4 or the TCR 4196_AV6_with_BV11-2 disclosed in US 2020/0277352. The TCR-expressing T cells were co-cultured for 18 hours (h) with T2 cells that expressed HLA-A2 and had been incubated with the WT epitope analogue HMTEVVRRX (SEQ ID NO: 117), wherein X at position 9 of SEQ ID NO: 117 is AABA, or p53R175H 9-amino acid epitope analogue at the concentration of peptides denoted Leydig 770310 HHS E-054-2023-0-PC-01 48 in Figures 1B-1E. T cell activation marker was measured by flow cytometry following the co-culture. The 4196-C TCR, 4196-D TCR, and 4196-E TCR all showed specificity for the mutant p53 epitope analogue. In contrast, the TCR 4196_AV6_with_BV11-2 showed significant wild-type epitope analogue recognition at the 1000 ng/mL and 100 ng/mL concentration. These results suggest that the 4196-C TCR, 4196-D TCR, and 4196-E TCR have superior specificity for the p53 R175H epitope analogue as compared to the TCR 4196_AV6_with_BV11-2. The titration curves of Figures 1B-1E were replotted for comparison, as shown in Figure 1F. Relative to the TCR 4196_AV6_with_BV11-2, the 4196-C TCR, 4196-D TCR, and 4196-E TCR showed better (4196-C TCR and 4196-E TCR) or equivalent (4196-D TCR) avidity for the mutant p53 R175H epitope analogue. These results suggest that 4196-C TCR, 4196-D TCR, and 4196-E TCR potentially have higher sensitivity for the p53 R175H antigen as compared to the TCR 4196_AV6_with_BV11-2. EXAMPLE 6 This example demonstrates that the 4196-C TCR, 4196-D TCR, and 4196-E TCR kill tumor cells. Effector cells were prepared by transducing healthy donor PBL with the retroviral vector encoding the 4196-C TCR, 4196-D TCR, or 4196-E TCR of Example 4. The anti- tumor activity of the 4196-C TCR, 4196-D TCR, and 4196-E TCR was determined using an INCUCYTE immune cell killing assay. TYK-nu human ovarian cancer cells (target cells) that naturally expressed p53 R175H and HLA-A2 were labeled with red fluorescent protein (RFP). Fifty thousand effector cells were co-cultured with ten thousand TYK-nu cells over 3 days. The number of viable RFP+ TYK-nu cells were counted at 4h intervals. The results are shown in Figure 1G. These data demonstrate that all of 4196-C TCR, 4196-D TCR, and 4196-E TCR can mediate significant tumor cell killing in vitro. EXAMPLE 7 This example demonstrates the specificity and avidity of the 4424-R175H TCR. Effector cells were healthy donor PBL transduced with the retroviral vector encoding the 4424-R175H TCR of Example 4. Target cells were monkey kidney COS7 cells that were engineered to express HLA-A2 and incubated with the WT peptide analogue HMTEVVRRX (SEQ ID NO: 117), wherein X at position 9 of SEQ ID NO: 117 is AABA or Leydig 770310 HHS E-054-2023-0-PC-01 49 the mutant p53 R175H peptide analogue (SEQ ID NO: 116), wherein X at position 9 of SEQ ID NO: 116 is AABA at the concentrations denoted in Fig.2B. Following 18 h co-culture, T cell activation marker 4-1BB expression was measured by flow cytometry. The results are shown in Fig.2B. The data showed that the 4424-R175H TCR is specific for mutant p53 R175H. Because COS7 cells used here only express HLA-A2, these data corroborate the HLA restriction of 4424-R175H TCR by HLA-A2. EXAMPLE 8 This example demonstrates a functional comparison of various TCRs reactive to p53 R175H. The ability of various TCRs targeting p53 R175H to kill tumor cells in vitro was compared using the INCUCYTE immune cell killing assay as described in Example 6. The TCR 4141-TCR1a2 disclosed in US 2022/0332785 was included as a reference. Twenty- thousand healthy donor T cells expressing the 4196-C TCR, 4196-D TCR, 4196-E TCR, or 4424-R175H TCR encoded by the respective retroviral vectors of Example 4 or 4141- TCR1a2 (effector cells) were co-cultured with ten thousand human ovarian cancer TYK-nu cells (target cells) for 72 h at an effector to target ratio of 2:1. Viable RFP+ TYK-nu cells were counted every 3 h. The results are shown in Figure 3. All of the T cell-treated conditions showed a significant delay in tumor growth relative to the tumor-only condition. The 4196-C TCR, 4196-D TCR, and 4424-R175H TCR showed a significant reduction in tumor growth as compared to the 4141-TCR1a2 TCR. EXAMPLE 9 This example demonstrates tumor cell recognition by the 4402-Y220C-B TCR, 4402-Y220C-C TCR, and 4402-Y220C-L TCR. Healthy donor PBL were independently transduced with the respective retroviral vectors of Example 4 encoding the 4402-Y220C-B TCR, 4402-Y220C-C TCR, or 4402- Y220C-L TCR (effector cells). Effector cells were co-cultured with target cells under conditions shown in Table 6. Leydig 770310 HHS E-054-2023-0-PC-01 50 6 Target cells Effector Cells Autologous breast tumor organoid cells 4402-Y220C-B TCR, 4402-Y220C-C TCR, , , , , ,
Figure imgf000052_0001
γ y p y. g ells from a different patient were included as a negative control. Autologous B cells pulsed with p53 Y220C 25-mer (DRNTFRHSVVVPCEPPEVGSDCTTI) (SEQ ID NO: 114) were included as a positive control. The 4402-Y220C-C TCR and 4402-Y220C-L TCR released IFN-γ when the autologous organoid tumor cells were pulsed with p53 Y220C 25-mer (DRNTFRHSVVVPCEPPEVGSDCTTI) (SEQ ID NO: 114). These results indicated that the tumor cells expressed HLA molecules restricting the 4402-Y220C-C TCR and 4402-Y220C- L TCR. The 4402-Y220C-B TCR did not release IFN-γ when the autologous organoid tumor cells were pulsed with p53 Y220C 25-mer (DRNTFRHSVVVPCEPPEVGSDCTTI) (SEQ ID NO: 114). The autologous organoid tumor cells exhibited HLA loss of heterozygosity, and they may have lost the HLA-DRB3*02:02 molecule restricting the 4402-Y220C-B TCR. Leydig 770310 HHS E-054-2023-0-PC-01 51 10 This example demonstrates the avidity and specificity of the 4402-Y220C-B TCR, 4402-Y220C-C TCR, and 4402-Y220C-L TCR. Healthy donor PBL were independently transduced with the retroviral vector of Example 4 encoding the 4402-Y220C-B TCR, 4402-Y220C-C TCR, or 4402-Y220C-L TCR or the 4343-D TCR disclosed in US 2023/0321240 (effector cells). Target cells were autologous B cells pulsed with WT (DRNTFRHSVVVPYEPPEVGSDCTTI) (SEQ ID NO: 115) or mutant (DRNTFRHSVVVPCEPPEVGSDCTTI) (SEQ ID NO: 114) peptides at the concentrations denoted in Figures 4A-4D. After the effector cells were co-cultured with the target cells, 4-1BB expression was measured by flow cytometry. The results are shown in Figures 4A-4D. The 4402-Y220C-B TCR, 4402-Y220C-C TCR, and 4402-Y220C-L TCR showed minimal WT p53 peptide recognition. In contrast, the 4343-D TCR showed significant WT peptide recognition. The 4402-Y220C-B TCR, 4402-Y220C-C TCR, and 4402-Y220C-L TCR showed superior mutant specificity relative to the 4343-D TCR. EXAMPLE 11 This example demonstrates that the 4402-Y220C-B TCR is HLA-DRB3*02:02- restricted. Healthy donor PBL were transduced with the retroviral vector of Example 4 encoding the 4402-Y220C-B TCR (effector cells). Target cells were COS7 cells independently transfected with the individual Class II HLA molecules expressed by Patient 4402 (Table 7). TABLE 7 HLA molecule(s) Transfected into Target Cells
Figure imgf000053_0001
Leydig 770310 HHS E-054-2023-0-PC-01 52
Figure imgf000054_0001
secretion was measured by ELISpot. 4402-Y220C-B TCR-expressing cells showed significant IFN- ^^ secretion when co-cultured with COS7 cells expressing HLA-DRB3*02:02. EXAMPLE 12 This example demonstrates differential autologous tumor organoid recognition by 4402-Y220C-C TCR and 4402-Y220C-L TCR. Healthy donor PBL were independently transduced with the retroviral vector of Example 4 encoding the 4402-Y220C-C TCR or 4402-Y220C-L TCR (effector cells). Target cells were autologous breast tumor organoid cells from Patient 4402 of passage 11 or passage 27. Target cells pulsed with DMSO served as a control. Effector cells were co-cultured with target cells. T cell activation was measured by 4-1BB upregulation. The results are shown in Figure 4E. The results showed that both TCRs only recognized the organoid cells of passage 27, suggesting that the organoid cells of passage 11 may have lost the HLA molecule restricting the 4402-Y220C-C TCR and 4402- Y220C-L TCR. EXAMPLE 13 This example demonstrates the differential expression of the HLA-DRB1*15:01 and DRB1*13:03 molecules by the autologous organoid cells of passage 11 and 27 of Example 12. The expression of HLA-DRB1*15:01 and DRB1*13:03 by the autologous organoid cells of passage 11 and 27 of Example 12 was measured by transcriptome analysis (RNA sequencing). The results are shown in Figure 4F. The Transcripts Per Kilobase Million (TPM) value of the two HLA molecules supports the loss of HLA-DRB1*13:03 in the organoid cells of passage 11. These results suggest that HLA-DRB1*13:03 is the restriction element for the 4402-Y220C-C TCR and 4402-Y220C-L TCR. Leydig 770310 HHS E-054-2023-0-PC-01 53 14 This example demonstrates that adoptive transfer of cells transduced with the retroviral vector encoding the 4196-C TCR of Example 4 reduce tumor size in tumor-bearing mice. NSG mice (immunocompromised) were subcutaneously injected with TYK-nu human ovarian cancer cells, which naturally expressed the p53 R175H mutation and HLA- A*02:01. Two weeks later, the tumor-bearing mice were randomized and treated with adoptive cell transfer of healthy donor T cells independently transduced with: 1. the retroviral vector encoding the 4196-C TCR of Example 4; 2. an irrelevant TCR control (i.e., 4259); 3. the TCR 4196_AV12-1_with_BV6-1 disclosed in US 2020/0277352; 4. the TCR 4196_AV38-1_with_BV10-3 disclosed in US 2020/0277352; 5. the TCR 4196_AV6_with_BV11-2 disclosed in US 2020/0277352; 6. the TCR 4141-TCR1a2 disclosed in US 2022/0332785; or 7. the 4141 IVS TCR disclosed in US Patent Application No.18/289,596. Two T-cell dose levels were tested: 2 million or 10 million T cells/mouse. The mean tumor size was measured at various time points up to 40 days after adoptive transfer of the transduced cells. The results obtained with the 2 million T cells/mouse dose are shown in Figure 6. As shown in Figure 6, only the 4196-C TCR showed statistical significance relative to 4259 TCR, the irrelevant TCR control. The results obtained with the 10 million T cells/mouse dose are shown in Figure 7. As shown in Figure 7, only the 4196-C TCR and the TCR 4196_AV6_with_BV11-2 showed statistical significance relative to 4259 TCR, the irrelevant TCR control. All references, including publications, patent applications, and patents, cited herein are hereby incorporated by reference to the same extent as if each reference were individually and specifically indicated to be incorporated by reference and were set forth in its entirety herein. The use of the terms “a” and “an” and “the” and “at least one” and similar referents in the context of describing the invention (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. The use of the term “at least one” Leydig 770310 HHS E-054-2023-0-PC-01 54 followed by a list of one or more items (for “at least one of A and B”) is to be construed to mean one item selected from the listed items (A or B) or any combination of two or more of the listed items (A and B), unless otherwise indicated herein or clearly contradicted by context. The terms “comprising,” “having,” “including,” and “containing” are to be construed as open-ended terms (i.e., meaning “including, but not limited to,”) unless otherwise noted. Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”) provided herein, is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention unless otherwise claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention. Preferred aspects of this invention are described herein, including the best mode known to the inventors for carrying out the invention. Variations of those preferred aspects may become apparent to those of ordinary skill in the art upon reading the foregoing description. The inventors expect skilled artisans to employ such variations as appropriate, and the inventors intend for the invention to be practiced otherwise than as specifically described herein. Accordingly, this invention includes all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Moreover, any combination of the above-described elements in all possible variations thereof is encompassed by the invention unless otherwise indicated herein or otherwise clearly contradicted by context.

Claims

Leydig 770310 HHS E-054-2023-0-PC-01 55 CLAIM(S): 1. An isolated or purified T cell receptor (TCR) having antigenic specificity for a human p53Y220C or human p53R175H amino acid sequence, wherein the TCR comprises the amino acid sequences of: (1) all of SEQ ID NOs: 2-7; (2) all of SEQ ID NOs: 16-21; (3) all of SEQ ID NOs: 30-35; (4) all of SEQ ID NOs: 44-49; (5) all of SEQ ID NOs: 58-63; (6) all of SEQ ID NOs: 72-77; or (7) all of SEQ ID NOs: 86-91. 2. The TCR of claim 1, wherein the TCR comprises the amino acid sequence(s) of: (1) SEQ ID NO: 8; (2) SEQ ID NO: 9; (3) both of SEQ ID NOs: 8 and 9; (4) SEQ ID NO: 10; (5) SEQ ID NO: 11; (6) both of SEQ ID NOs: 10 and 11; (7) SEQ ID NO: 22; (8) SEQ ID NO: 23; (9) both of SEQ ID NOs: 22 and 23; (10) SEQ ID NO: 24; (11) SEQ ID NO: 25; (12) both of SEQ ID NOs: 24 and 25; (13) SEQ ID NO: 36; (14) SEQ ID NOs: 37; (15) both of SEQ ID NOs: 36 and 37; (16) SEQ ID NO: 38; (17) SEQ ID NO: 39; (18) both of SEQ ID NOs: 38 and 39; (19) SEQ ID NO: 50; Leydig 770310 HHS E-054-2023-0-PC-01 56 (20) SEQ ID NO: 51; (21) both of SEQ ID NOs: 50 and 51; (22) SEQ ID NO: 52; (23) SEQ ID NO: 53; (24) both of SEQ ID NOs: 52 and 53; (25) SEQ ID NO: 64; (26) SEQ ID NO: 65; (27) both of SEQ ID NOs: 64 and 65; (28) SEQ ID NO: 66; (29) SEQ ID NO: 67; (30) both of SEQ ID NOs: 66 and 67; (31) SEQ ID NO: 78; (32) SEQ ID NO: 79; (33) both of SEQ ID NOs: 78 and 79; (34) SEQ ID NO: 80; (35) SEQ ID NO: 81; (36) both of SEQ ID NOs: 80 and 81; (37) SEQ ID NO: 92; (38) SEQ ID NO: 93; (39) both of SEQ ID NOs: 92 and 93; (40) SEQ ID NO: 94; (41) SEQ ID NO: 95; or (42) both of SEQ ID NO: 94 and 95. 3. The TCR of claim 1 or 2, wherein the TCR comprises the amino acid sequences of: (1) SEQ ID NO: 12; (2) SEQ ID NO: 13; (3) both of SEQ ID NOs: 12 and 13; (4) SEQ ID NO: 14; (5) SEQ ID NO: 15; (6) both of SEQ ID NOs: 14 and 15; (7) SEQ ID NO: 26; Leydig 770310 HHS E-054-2023-0-PC-01 57 (8) SEQ ID NO: 27; (9) both of SEQ ID NOs: 26 and 27; (10) SEQ ID NO: 28; (11) SEQ ID NO: 29; (12) both of SEQ ID NOs: 28 and 29; (13) SEQ ID NO: 40; (14) SEQ ID NO: 41; (15) both of SEQ ID NOs: 40 and 41; (16) SEQ ID NO: 42; (17) SEQ ID NO: 43; (18) both of SEQ ID NOs: 42 and 43; (19) SEQ ID NO: 54; (20) SEQ ID NO: 55; (21) both of SEQ ID NOs: 54 and 55; (22) SEQ ID NO: 56; (23) SEQ ID NO: 57; (24) both of SEQ ID NOs: 56 and 57; (25) SEQ ID NO: 68; (26) SEQ ID NO: 69; (27) both of SEQ ID NOs: 68 and 69; (28) SEQ ID NO: 70; (29) SEQ ID NO: 71; (30) both of SEQ ID NOs: 70 and 71; (31) SEQ ID NO: 82; (32) SEQ ID NO: 83; (33) both of SEQ ID NO: 82 and 83; (34) SEQ ID NO: 84; (35) SEQ ID NO: 85; (36) both of SEQ ID NO: 84 and 85; (37) SEQ ID NO: 96; (38) SEQ ID NO: 97; (39) both of SEQ ID NO: 96 and 97; (40) SEQ ID NO: 98; Leydig 770310 HHS E-054-2023-0-PC-01 58 (41) SEQ ID NO: 99; or (42) both of SEQ ID NO: 98 and 99. 4. The TCR of any one of claims 1-3, wherein the human p53Y220C amino acid sequence is DRNTFRHSVVVPCEPPEVGSDCTTI (SEQ ID NO: 114). 5. The TCR of any one of claims 1-4, wherein the TCR does not have antigenic specificity for the wild-type human p53 amino acid sequence of DRNTFRHSVVVPYEPPEVGSDCTTI (SEQ ID NO: 115). 6. The TCR of any one of claims 1-3, wherein the human p53R175H amino acid sequence is HMTEVVRHC (SEQ ID NO: 112). 7. The TCR of any one of claims 1-3 and 6, wherein the TCR does not have antigenic specificity for the wild-type human p53 amino acid sequence of HMTEVVRRC (SEQ ID NO: 113). 8. An isolated or purified polypeptide comprising a functional portion of the TCR of any one of claims 1-7, wherein the polypeptide comprises the amino acid sequences of: (1) all of SEQ ID NOs: 2-7; (2) all of SEQ ID NOs: 16-21; (3) all of SEQ ID NOs: 30-35; (4) all of SEQ ID NOs: 44-49; (5) all of SEQ ID NOs: 58-63; (6) all of SEQ ID NOs: 72-77; or (7) all of SEQ ID NOs: 86-91. 9. The polypeptide of claim 8, wherein the polypeptide comprises the amino acid sequences of: (1) SEQ ID NO: 8; (2) SEQ ID NO: 9; (3) both of SEQ ID NOs: 8 and 9; (4) SEQ ID NO: 10; Leydig 770310 HHS E-054-2023-0-PC-01 59 (5) SEQ ID NO: 11; (6) both of SEQ ID NOs: 10 and 11; (7) SEQ ID NO: 22; (8) SEQ ID NO: 23; (9) both of SEQ ID NOs: 22 and 23; (10) SEQ ID NO: 24; (11) SEQ ID NO: 25; (12) both of SEQ ID NOs: 24 and 25; (13) SEQ ID NO: 36; (14) SEQ ID NOs: 37; (15) both of SEQ ID NOs: 36 and 37; (16) SEQ ID NO: 38; (17) SEQ ID NO: 39; (18) both of SEQ ID NOs: 38 and 39; (19) SEQ ID NO: 50; (20) SEQ ID NO: 51; (21) both of SEQ ID NOs: 50 and 51; (22) SEQ ID NO: 52; (23) SEQ ID NO: 53; (24) both of SEQ ID NOs: 52 and 53; (25) SEQ ID NO: 64; (26) SEQ ID NO: 65; (27) both of SEQ ID NOs: 64 and 65; (28) SEQ ID NO: 66; (29) SEQ ID NO: 67; (30) both of SEQ ID NOs: 66 and 67; (31) SEQ ID NO: 78; (32) SEQ ID NO: 79; (33) both of SEQ ID NOs: 78 and 79; (34) SEQ ID NO: 80; (35) SEQ ID NO: 81; (36) both of SEQ ID NOs: 80 and 81; (37) SEQ ID NO: 92; Leydig 770310 HHS E-054-2023-0-PC-01 60 (38) SEQ ID NO: 93; (39) both of SEQ ID NOs: 92 and 93; (40) SEQ ID NO: 94; (41) SEQ ID NO: 95; or (42) both of SEQ ID NO: 94 and 95. 10. The polypeptide of claim 8 or 9, wherein the polypeptide comprises the amino acid sequences of: (1) SEQ ID NO: 12; (2) SEQ ID NO: 13; (3) both of SEQ ID NOs: 12 and 13; (4) SEQ ID NO: 14; (5) SEQ ID NO: 15; (6) both of SEQ ID NOs: 14 and 15; (7) SEQ ID NO: 26; (8) SEQ ID NO: 27; (9) both of SEQ ID NOs: 26 and 27; (10) SEQ ID NO: 28; (11) SEQ ID NO: 29; (12) both of SEQ ID NOs: 28 and 29; (13) SEQ ID NO: 40; (14) SEQ ID NO: 41; (15) both of SEQ ID NOs: 40 and 41; (16) SEQ ID NO: 42; (17) SEQ ID NO: 43; (18) both of SEQ ID NOs: 42 and 43; (19) SEQ ID NO: 54; (20) SEQ ID NO: 55; (21) both of SEQ ID NOs: 54 and 55; (22) SEQ ID NO: 56; (23) SEQ ID NO: 57; (24) both of SEQ ID NOs: 56 and 57; (25) SEQ ID NO: 68; Leydig 770310 HHS E-054-2023-0-PC-01 61 (26) SEQ ID NO: 69; (27) both of SEQ ID NOs: 68 and 69; (28) SEQ ID NO: 70; (29) SEQ ID NO: 71; (30) both of SEQ ID NOs: 70 and 71; (31) SEQ ID NO: 82; (32) SEQ ID NO: 83; (33) both of SEQ ID NO: 82 and 83; (34) SEQ ID NO: 84; (35) SEQ ID NO: 85; (36) both of SEQ ID NO: 84 and 85; (37) SEQ ID NO: 96; (38) SEQ ID NO: 97; (39) both of SEQ ID NO: 96 and 97; (40) SEQ ID NO: 98; (41) SEQ ID NO: 99; or (42) both of SEQ ID NO: 98 and 99. 11. An isolated or purified protein comprising first and second polypeptide chains, wherein: (1) the first polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 2-4; (2) the second polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 5-7; (3) the first polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 2-4 and the second polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 5-7; (4) the first polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 16-18; (5) the second polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 19-21; Leydig 770310 HHS E-054-2023-0-PC-01 62 (6) the first polypeptide chain the amino acid sequences of all of SEQ ID NOs: 16-18 and the second polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 19-21; (7) the first polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 30-32; (8) the second polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 33-35; (9) the first polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 30-32 and the second polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 33-35; (10) the first polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 44-46; (11) the second polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 47-49; (12) the first polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 44-46 and the second polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 47-49; (13) the first polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 58-60; (14) the second polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 61-63; (15) the first polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 58-60 and the second polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 61-63; (16) the first polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 72-74; (17) the second polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 75-77; (18) the first polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 72-74 and the second polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 75-77; (19) the first polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 86-88; Leydig 770310 HHS E-054-2023-0-PC-01 63 (20) the second polypeptide chain the amino acid sequences of all of SEQ ID NOs: 89-91; or (21) the first polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 86-88 and the second polypeptide chain comprises the amino acid sequences of all of SEQ ID NOs: 89-91. 12. The protein of claim 11, wherein: (1) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 8; (2) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 9; (3) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 8 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 9; (4) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 10; (5) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 11; (6) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 10 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 11; (7) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 22; (8) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 23; (9) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 22 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 23; (10) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 24; (11) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 25; (12) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 24 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 25; (13) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 36; (14) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 37; Leydig 770310 HHS E-054-2023-0-PC-01 64 (15) the first polypeptide chain the amino acid sequence of SEQ ID NO: 36 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 37; (16) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 38; (17) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 39; (18) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 38 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 39; (19) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 50; (20) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 51; (21) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 50 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 51; (22) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 52; (23) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 53; (24) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 52 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 53; (25) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 64; (26) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 65; (27) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 64 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 65; (28) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 66; (29) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 67; (30) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 66 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 67; Leydig 770310 HHS E-054-2023-0-PC-01 65 (31) the first polypeptide chain the amino acid sequence of SEQ ID NO: 78; (32) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 79; (33) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 78 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 79; (34) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 80; (35) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 81; (36) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 80 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 81; (37) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 92; (38) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 93; (39) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 92 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 93; (40) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 94; (41) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 95; or (42) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 94 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 95. 13. The protein of claim 11 or 12, wherein: (1) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 12; (2) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 13; (3) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 12 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 13; (4) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 14; Leydig 770310 HHS E-054-2023-0-PC-01 66 (5) the second polypeptide chain the amino acid sequence of SEQ ID NO: 15; (6) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 14 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 15; (7) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 26; (8) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 27; (9) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 26 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 27; (10) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 28; (11) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 29; (12) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 28 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 29; (13) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 40; (14) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 41; (15) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 40 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 41; (16) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 42; (17) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 43; (18) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 42 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 43; (19) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 54; (20) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 55; (21) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 54 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 55; Leydig 770310 HHS E-054-2023-0-PC-01 67 (22) the first polypeptide chain the amino acid sequence of SEQ ID NO: 56; (23) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 57; (24) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 56 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 57; (25) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 68; (26) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 69; (27) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 68 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 69; (28) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 70; (29) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 71; (30) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 70 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 71; (31) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 82; (32) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 83; (33) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 82 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 83; (34) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 84; (35) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 85; (36) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 84 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 85; (37) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 96; Leydig 770310 HHS E-054-2023-0-PC-01 68 (38) the second polypeptide chain the amino acid sequence of SEQ ID NO: 97; (39) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 96 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 97; (40) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 98; (41) the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 99; or (42) the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 98 and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 99. 14. A bispecific engager TCR fusion protein comprising (i) the TCR according to any one of claims 1-7, the polypeptide according to any one of claims 8-10, or the protein according to any one of claims 11-13 and (ii) an anti-CD3 effector. 15. An isolated or purified nucleic acid comprising a nucleotide sequence encoding the TCR of any one of claims 1-7, the polypeptide of any one of claims 8-10, the protein of any one of claims 11-14. 16. An isolated or purified nucleic acid comprising, from 5’ to 3’, a first nucleic acid sequence and a second nucleotide sequence, wherein the first and second nucleotide sequence, respectively, encode the amino sequences of SEQ ID NOs: 8 and 9; 9 and 8; 10 and 11; 11 and 10; 12 and 13; 13 and 12; 14 and 15; 15 and 14; 22 and 23; 23 and 22; 24 and 25; 25 and 24; 26 and 27; 27 and 26; 28 and 29; 29 and 28; 36 and 37; 37 and 36; 38 and 39; 39 and 38; 40 and 41; 41 and 40; 42 and 43; 43 and 42; 50 and 51; 51 and 50; 52 and 53; 53 and 52; 54 and 55; 55 and 54; 56 and 57; 57 and 56; 64 and 65; 65 and 64; 66 and 67; 67 and 66; 68 and 69; 69 and 68; 70 and 71; 71 and 70; 78 and 79; 79 and 78; 80 and 81; 81 and 80; 82 and 83; 83 and 82; 84 and 85; 85 and 84; 92 and 93; 93 and 92; 94 and 95; 95 and 94; 96 and 97; 97 and 96; 98 and 99; or 99 and 98. 17. The isolated or purified nucleic acid of claim 16, further comprising a third nucleotide acid sequence interposed between the first and second nucleotide sequence, wherein the third nucleotide sequence encodes a cleavable linker peptide. Leydig 770310 HHS E-054-2023-0-PC-01 69 18. The isolated or purified nucleic acid of claim 17, wherein the cleavable linker peptide comprises the amino acid sequence of RAKRSGSGATNFSLLKQAGDVEENPGP (SEQ ID NO: 103). 19. A recombinant expression vector comprising the nucleic acid of any one of claims 15-18. 20. The recombinant expression vector of claim 19, which is a transposon or a lentiviral vector. 21. An isolated or purified TCR, polypeptide, or protein encoded by the nucleic acid of any one of claims 15-18 or the vector of claim 19 or 20. 22. An isolated or purified TCR, polypeptide, or protein that results from expression of the nucleic acid of any one of claims 15-18 or the vector of claim 19 or 20 in a cell. 23. A method of producing a host cell expressing a TCR that has antigenic specificity for a human p53Y220C or human p53R175H amino acid sequence, the method comprising contacting a cell in vitro with the vector of claim 19 or 20 under conditions that allow introduction of the vector into the cell. 24. An isolated or purified host cell comprising the nucleic acid of any one of claims 15-18 or the recombinant expression vector of claim 19 or 20. 25. The host cell of claim 24, wherein the cell is a human lymphocyte. 26. The host cell of claim 24, wherein the cell is selected from the group consisting of a T cell, a natural killer T (NKT) cell, an invariant natural killer T (iNKT) cell, a natural killer (NK) cell, a macrophage, a pluripotent cell, and a multipotent cell. 27. An isolated or purified population of cells comprising the host cell of any one of claims 24-26. Leydig 770310 HHS E-054-2023-0-PC-01 70 28. A method of producing the TCR of any one of claims 1-7, 21, or 22, the polypeptide of any one of claims 8-10, 21, or 22, or the protein of any one of claims 11-14, 21, or 22, the method comprising culturing the host cell of any one of claims 24-26, or the population of host cells of claim 27, so that the TCR, polypeptide, or protein is produced. 29. A pharmaceutical composition comprising (a) the TCR of any one of claims 1-7, 21, or 22, the polypeptide of any one of claims 8-10, 21, or 22, the protein of any one of claims 11-14, 21, or 22, the nucleic acid of any one of claims 15-18, the recombinant expression vector of claim 19 or 20, the host cell of any one of claims 24-26, or the population of host cells of claim 27 and (b) a pharmaceutically acceptable carrier. 30. A method of detecting the presence of cancer in mammal, the method comprising: (a) contacting a sample comprising cells of the cancer with the TCR of any one of claims 1-7, 21, or 22, the polypeptide of any one of claims 8-10, 21, or 22, the protein of any one of claims 11-14, 21, or 22, the nucleic acid of any one of claims 15-18, the recombinant expression vector of claim 19 or 20, the host cell of any one of claims 24-26, the population of host cells of claim 27, or the pharmaceutical composition of claim 29, thereby forming a complex; and (b) detecting the complex, wherein detection of the complex is indicative of the presence of cancer in the mammal. 31. The TCR of any one of claims 1-7, 21, or 22, the polypeptide of any one of claims 8-10, 21, or 22, the protein of any one of claims 11-14, 21, or 22, the nucleic acid of any one of claims 15-18, the recombinant expression vector of claim 19 or 20, the host cell of any one of claims 24-26, the population of host cells of claim 27, or the pharmaceutical composition of claim 29, for use in the inducement of an immune response against cancer in a mammal. 32. The TCR of any one of claims 1-7, 21, or 22, the polypeptide of any one of claims 8-10, 21, or 22, the protein of any one of claims 11-14, 21, or 22, the nucleic acid of Leydig 770310 HHS E-054-2023-0-PC-01 71 any one of claims 15-18, the recombinant vector of claim 19 or 20, the host cell of any one of claims 24-26, the population of host cells of claim 27, or the pharmaceutical composition of claim 29, for use in the treatment or prevention of cancer in a mammal. 33. The population of host cells for the use of claim 31 or 32, wherein the population of cells is autologous to the mammal. 34. The population of host cells for the use of claim 31 or 32, wherein the population of cells is allogeneic to the mammal. 35. The TCR, polypeptide, protein, nucleic acid, recombinant expression vector, host cell, population of host cells, or pharmaceutical composition for the use of any one of claims 30-34, wherein the cancer is an epithelial cancer. 36. The TCR, polypeptide, protein, nucleic acid, recombinant expression vector, host cell, population of host cells, or pharmaceutical composition for the use of any one of claims 30-34, wherein the cancer is cholangiocarcinoma, melanoma, colon cancer, rectal cancer, ovarian cancer, endometrial cancer, non-small cell lung cancer (NSCLC), glioblastoma, uterine cervical cancer, head and neck cancer, breast cancer, pancreatic cancer, or bladder cancer. 37. The TCR, polypeptide, protein, nucleic acid, recombinant expression vector, host cell, population of host cells, or pharmaceutical composition for the use of any one of claims 30-36, wherein the cancer is known to comprise an Y220C or R175H mutation in human p53.
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