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WO2024167312A1 - Protéine dkk-1 présente de manière spécifique sur des lymphocytes t régulateurs et son utilisation - Google Patents

Protéine dkk-1 présente de manière spécifique sur des lymphocytes t régulateurs et son utilisation Download PDF

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WO2024167312A1
WO2024167312A1 PCT/KR2024/001828 KR2024001828W WO2024167312A1 WO 2024167312 A1 WO2024167312 A1 WO 2024167312A1 KR 2024001828 W KR2024001828 W KR 2024001828W WO 2024167312 A1 WO2024167312 A1 WO 2024167312A1
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cancer
dkk
fibrosis
present
cells
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이래경크리스티나
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Jrd Science Inc
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • 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
    • C07K14/4702—Regulators; Modulating activity
    • C07K14/4703—Inhibitors; Suppressors
    • 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/30—Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto

Definitions

  • the present invention relates to a diagnostic composition for diagnosing fibrosis or cancer, a diagnostic kit, a method for providing information, and a pharmaceutical composition for preventing or treating fibrosis or cancer, comprising a formulation for measuring the expression level of DKK-1 (Dickkopf-1) protein or a gene encoding the same, which is specifically present on the surface of regulatory T cells (Treg cells) among immune cells; and a regulatory T cell comprising an extracellular surface protein of DKK-1, a fusion protein comprising an extracellular surface protein of DKK-1 and an immunoglobulin Fc region, an antibody or antigen-binding fragment specifically binding to the extracellular surface protein of DKK-1, or an epitope, and a DKK-1 inhibitor.
  • DKK-1 Dickkopf-1
  • Treg cells regulatory T cells
  • a regulatory T cell comprising an extracellular surface protein of DKK-1, a fusion protein comprising an extracellular surface protein of DKK-1 and an immunoglobulin Fc region, an antibody or antigen-binding fragment specifically
  • T cells are immune cells that are produced in the bone marrow and mature in the thymus, and they function to regulate the antibody production of B cells or to regulate the functions of innate immune cells.
  • the T cells are also called T lymphocytes and not only act as helpers for other immune cells, but also directly destroy invading substances.
  • immune cells One of the most important functions of immune cells is to suppress the immune response to antigenic substances that constitute the self surrounding the immune cell, while recognizing and inducing an immune response to non-self antigenic substances.
  • immune cells induce non-responsiveness to self-antigens through the death of cells that recognize self, mutation induction of receptors specific to self-antigens, or inactivation of immune cells that recognize self-antigens. This is called immunologic unresponsiveness or tolerance. If this self-tolerance fails, an immune response to self-antigens can be induced, resulting in disease, which is called an autoimmune disease.
  • the regulatory T cells control the function of classical T cells to induce excessive immune responses or self-tolerance.
  • the function and number of regulatory T cells are significantly reduced in autoimmune diseases and chronic inflammatory diseases, and thus the regulatory T cells do not perform their function properly. Therefore, restoring the function and number of regulatory T cells to a normal level in patients with autoimmune diseases and chronic inflammatory diseases may be one of the treatments for the diseases.
  • CD25, CTLA4, CD38, CD62L, GITR, CD45RB, etc. have been suggested through research as cell surface proteins that can target regulatory T cells, and animal and clinical trials have also been conducted, but a cell surface protein that can exclusively target regulatory T cells has not been discovered to date.
  • One object of the present invention is to provide a composition for diagnosing fibrosis or cancer, comprising an antibody or antigen-binding fragment that specifically binds to a DKK-1 extracellular surface protein or epitope.
  • Another object of the present invention is to provide a diagnostic kit for fibrosis or cancer comprising the composition.
  • Another object of the present invention is to provide a method for providing information on fibrosis or cancer by measuring the expression level of the extracellular surface protein or epitope of DKK-1 present in regulatory T cells using an antibody or antigen-binding fragment that specifically binds to the surface protein or epitope.
  • Another object of the present invention is to provide a diagnostic device for fibrosis or cancer, including: (a) a measuring unit for measuring the expression level of DKK-1 or a gene encoding the protein for a biological sample obtained from a target subject; and (b) a detection unit for outputting the presence or absence of cancer from the expression level of the gene or the protein encoded by the gene measured by the measuring unit.
  • Another object of the present invention is to provide a pharmaceutical composition for preventing or treating fibrosis or cancer, comprising as an active ingredient a fusion protein comprising LRP-5, 6 and an immunoglobulin Fc region, an extracellular surface protein of DKK-1, or an antibody or antigen-binding fragment specifically binding to an epitope, or a DKK-1 inhibitor.
  • DKK-1 or “DKK1”, known as a representative negative regulator of the Wnt signaling pathway, competitively binds to LRP (Low density lipoprotein receptor-related protein) 5/6 with Wnt, or ultimately induces B-Catenin degradation and TCF-1 inactivation through endocytosis of the receptor.
  • DKK-1 is the most studied protein among the four DKK families and has a size of approximately 26 kDa composed of 266 amino acids. Looking at the structure of the protein, it interacts with domains of various receptors including LRP6 through two cysteine rich domains (CRD).
  • the currently known receptors for DKK-1 include LRP5/6, Kremen 1/2, and CKAP4, and it induces various cell signals through the above receptors.
  • DDK-1 is closely related to cell proliferation through Wnt signaling and CKAP4-PI3K-AKT signaling
  • high expression of DDK-1 in cancer indicates a negative prognosis, although the degree varies depending on the type of cancer.
  • many studies are being conducted on cancer immunotherapy targeting DDK-1.
  • recent papers have revealed the role of DDK-1 in various immune responses, and the results of studies on the influence of DDK-1 in autoimmune diseases can be confirmed.
  • DDK-1 DDK-1 secretion from platelets
  • MAPK and mTOR signaling The increase in DDK-1 under physiological conditions generates Th2 cells through the induction of C-Maf and Gata3, and causes the expression of Th2 cell-related cytokines through p38 MAPK and SGK-1. That is, DDK-1 is preferentially involved in functions associated with Th2 compared to other effector T cells.
  • DDK-1 inhibits the expression of IFN-r under Th1 polarization conditions, and simultaneously increases the expression of cytokines such as GATA3 and IL-4, IL-5, IL-10, and IL-13.
  • cytokines such as GATA3 and IL-4, IL-5, IL-10, and IL-13.
  • Foxp3 the expression of Foxp3 is down-regulated.
  • platelets are known to be a major source of TGF-B1, and in the inflammatory response environment created through the above infection, it is presumed that DDK-1, together with CXCL4, a ligand produced by platelets, lowers Foxp3 expression, thereby inhibiting TGF-B-mediated Foxp3+ Treg cell production and allowing Th2 to be properly activated.
  • nTreg cells have a high level of DDK-1 mRNA expression compared to na ⁇ ve T cells or effector CD4 T cells.
  • the generally known DDK-1 is known to bind to the receptor as a ligand in a soluble form and play its role, but DDK-1 of nTreg is expressed on the surface, and about 6% of DDK-1 is expressed in the resting state, and the expression rate increases to 14.1% when activated.
  • DDK-1 is thought to play an important role in the suppressive function of Treg.
  • Treg did not suppress the proliferation of effector CD4 T cells.
  • Treg cells from mice with reduced DDK-1 expression by more than 90% did not suppress the proliferation of effector CD4 T cells as effectively as Wild Type Treg, ultimately resulting in the development of severe colitis.
  • DDK-1 in Treg it was shown that it is expressed independently of the Wnt pathway, mTOR, and Mevalonate pathways, and that the MAPK pathway is required for activated Treg to express DDK-1.
  • DKK1 which acts as an antagonist of Wnt, indicates the possibility of erosion due to increased bone resorption when measured at high concentrations in the blood, whereas low concentrations are considered to be new bone formation.
  • RA rheumatoid arthritis
  • DKK-1 protein is highly expressed in human regulatory T cells and its expression is reduced in iTreg.
  • the DKK-1 protein is an anchored protein existing outside the human Treg cell membrane, and can then be separated into the blood and exist in a soluble state.
  • the pharmaceutical composition provided by the present invention can exert an effect on fibrosis or cancer by regulating the immune response by inhibiting the function of DKK-1.
  • the DKK-1 is one of the DKK family genes DKK1, DKK2, DKK3, and DKK4, and encodes a secreted protein.
  • the DKK protein is generally composed of 255 to 350 amino acids, and in the case of DKK-1, has a size of 24 KDa to 29 KDa, and the N-terminus exists in a form in which a sugar is bound (glycosylated).
  • the DKK-1 can be represented by the amino acid sequence of SEQ ID NO: 1, and can be encoded by the nucleotide represented by SEQ ID NO: 2 (Table 1).
  • dickkopf-1 (DKK-1) amino acid sequence MMALGAAGAT RVFVAMVAAA LGGHPLLGVS ATLNSVLNSN AIKNLPPPLG GAAGHPGSAV SAAPGILYPG GNKYQTIDNY QPYPCAEDEE CGTDEYCASP TRGGDAGVQI CLACRKRRKR CMRHAMCCPG NYCKNGICVS SDQNHFRGEI EETITESFGN DHSTLDGYSR RTTLSSKMYH TKGQEGSVCL RSSDCASGLC CARHFWS KIC KPVLKEGQVC TKHRRKGSHG LEIFQRCYCG EGLSCRIQKD HHQASNSSRL HTCQRH (SEQ ID NO: 1) dickkopf-1 (DKK-1) mRNA, complete cds (GenBank: AF177394.2) sequence atgatggctc tgggcgcagc gggagctacc cgggtctttg tcgcgatgg
  • the DKK-1 may play an important role in the development of the heart, head, and hands during embryonic development by competitively inhibiting the Wnt signal by binding more strongly to the receptor than a ligand such as Wnt3a in the Wnt signaling system involved in cell proliferation and wound healing.
  • the DKK-1 may not be secreted outside the cell as in general cases, but may be present on the surface of immune cells, particularly regulatory T cells. Through this, immune cells may help the function of regulatory T cells for self-tolerance.
  • the regulatory T cell extracellular surface protein of the DKK-1 is represented by SEQ ID NO: 3 (Table 2).
  • the regulatory T cell extracellular surface protein of DKK-1 provided in the present invention can reduce the activity of regulatory T cells by interacting with a ligand present in effector T cells.
  • the epitope of DKK1 which is a surface protein of regulatory T cells, is represented by any one of SEQ ID NOS: 4 to 36 (Table 3).
  • the "epitope” or “epitope” refers to a portion of an antigen molecule that binds to an antibody, and refers to a portion that can be recognized by an antibody.
  • antibodies do not recognize the entire antigen molecule, but only a specific portion, and even if it is the same antigen molecule, if the type of antibody is different, it can recognize different antigenic determinant portions.
  • the antigenic determinant includes an antigenic determinant of an antibody that can effectively bind to a remaining portion of a domain even when a portion of a DKK1 protein exposed to the outside of the cell is partially cleaved and released (secreted) to the outside of the cell.
  • the polypeptide that is the antigenic determinant may include all continuous or discontinuous sequences of a portion that can be bound to the DKK-1 protein according to the present invention by an antibody.
  • the polypeptide fragment that is the antigenic determinant of DKK-1 which is the surface protein of regulatory T cells provided in the present invention, may interact with a ligand present in effector T cells to reduce the activity of regulatory T cells.
  • a polynucleotide encoding an extracellular surface protein of DKK-1 represented by SEQ ID NO: 3 provided in the present invention, or an antigenic determinant of DKK-1 of any one of SEQ ID NOs: 4 to 36.
  • an expression vector into which a polynucleotide provided by the present invention is inserted is provided.
  • the "vector” is a nucleic acid molecule capable of transporting another nucleic acid to which a nucleic acid molecule is linked.
  • a vector which refers to a circular double-stranded DNA to which additional DNA segments can be ligated.
  • a phage vector Another type of vector is a viral vector, to which additional DNA segments can be ligated to the viral genome.
  • Some vectors are capable of autonomous replication in the host cell into which they are introduced (e.g., a bacterial vector is an episomal mammalian vector having a bacterial origin of replication).
  • vectors e.g., a non-episomal mammalian vector
  • a non-episomal mammalian vector can be integrated into the genome of a host cell upon introduction into the host cell, thereby replicating along with the host genome.
  • some vectors can direct the expression of genes to which they are linked in an operational manner.
  • Such vectors are referred to herein as "recombinant expression vectors" or simply "expression vectors”.
  • expression vectors useful in recombinant DNA techniques are often in the form of plasmids.
  • plasmid and “vector” may be used interchangeably, as the plasmid is the most commonly used form of vector.
  • specific examples of the expression vector may be selected from the group consisting of commercially widely used pCDNA vectors, F, R1, RP1, Col, pBR322, ToL, Ti vectors; cosmids; phages such as lambda, lambdoid, M13, Mu, p1 P22, Q ⁇ , T-even, T2, T3, T7; and plant viruses, but are not limited thereto, and all expression vectors known to those skilled in the art as expression vectors can be used in the present invention, and when selecting an expression vector, it depends on the properties of the target host cell.
  • the vector When introducing a vector into a host cell, it may be performed by calcium phosphate transfection, viral infection, DEAE-dextran controlled transfection, lipofectamine transfection, or electroporation, but is not limited thereto, and those skilled in the art can select and use an introduction method appropriate for the expression vector and host cell to be used.
  • the vector contains one or more selection markers, but is not limited thereto, and selection is possible based on whether or not the product is produced using a vector that does not contain a selection marker.
  • the selection marker is selected by the desired host cell, and this is done using a method already known to those skilled in the art, so the present invention is not limited thereto.
  • a tag sequence can be inserted into an expression vector and fused thereto.
  • the tag includes, but is not limited to, a hexa-histidine tag, a hemagglutinin tag, a myc tag, or a flag tag, and any tag known to those skilled in the art that facilitates purification can be used in the present invention.
  • the expression vector provided in the present invention provides a host cell line transfected with the expression vector.
  • the "host cell” in the present invention includes an individual cell or cell culture that can be or has been the recipient of the vector(s) for incorporation of the polypeptide insert.
  • the host cell includes progeny of a single host cell, which progeny may not necessarily be completely identical (either morphologically or in genomic DNA complement) to the original parent cell due to natural, accidental or deliberate mutation.
  • the host cell includes a cell that has been transfected in vivo with the polypeptide(s) of the present disclosure.
  • the host cell may include a cell of mammalian, plant, insect, fungal or cellular origin, and for example, bacterial cells such as Escherichia coli, Streptomyces, and Salmonella Typhimurium; fungal cells such as yeast cells and Pichia pastoris; insect cells such as Drozophylla and Spodoptera Sf9 cells; animal cells such as CHO (Chinese hamster ovary cells), SP2/0 (mouse myeloma), human lymphoblastoid, COS, NSO (mouse myeloma), 293T, Bow melanoma cells, HT-1080, BHK (Baby Hamster Kidney cells), HEK (Human Embryonic Kidney cells), or PERC.6 (human retinal cells); Or, it may be a plant cell, but is not limited thereto, and any cell that can be used as a host cell line known to those skilled in the art may be used.
  • bacterial cells such as Escherichia
  • a regulatory T cell comprising the extracellular surface protein of the DKK-1 of the present invention or an epitope of the DKK-1 is provided.
  • Regulatory T cells expressing the extracellular surface protein of the DKK-1 of the present invention or an antigenic determinant thereof on their surface can suppress the cellular activity of effector T cells through interaction with them and suppress an immune response.
  • a pharmaceutical composition for preventing or treating fibrosis or cancer comprising as an effective ingredient an extracellular surface protein of the DKK-1 of the present invention or a regulatory T cell comprising an epitope of the DKK-1.
  • Regulatory T cells comprising the extracellular surface protein of the DKK1 of the present invention can suppress the cell activity of effector T cells through interaction with them and suppress immune responses.
  • regulatory T cells expressing DKK-1 protein on their cell surface can effectively prevent, improve or treat immune-related diseases, such as fibrosis or cancer.
  • a pharmaceutical composition for preventing or treating fibrosis or cancer comprising the DKK-1 extracellular surface protein or the antigenic determinant of the present invention as an effective ingredient.
  • the DKK-1 extracellular surface protein or the antigen determinant fragment provided in the present invention interacts with a ligand present in effector T cells to inhibit the activity of regulatory T cells, thereby effectively preventing, improving or treating immune-related diseases, such as fibrosis or cancer.
  • the "cancer” refers to or represents a physiological condition characterized by uncontrolled cell growth typically in mammals.
  • the cancer to be prevented, improved or treated in the present invention may be a solid tumor formed by a mass caused by abnormal cell growth in a solid organ, and depending on the site of the solid organ, may be gastric cancer, liver cancer, glioma, ovarian cancer, colon cancer, head and neck cancer, bladder cancer, renal cell carcinoma, breast cancer, metastatic cancer, prostate cancer, pancreatic cancer, melanoma or lung cancer, and preferably may be melanoma, but is not limited thereto.
  • the fibrosis may be selected from the group consisting of fibrosis occurring in at least one organ selected from the group consisting of kidney, liver, lung, heart, bone or bone marrow and skin, but is not limited thereto.
  • a pharmaceutical composition for preventing or treating fibrosis or cancer comprising an antibody specific for the DKK1 extracellular surface protein or the antigenic determinant of the present invention as an active ingredient.
  • the DKK1 extracellular surface protein may be present on the surface of regulatory T cells.
  • the antibody can effectively prevent, improve or treat fibrosis or cancer by suppressing the activity of regulatory T cells.
  • the term "antibody” is a term known in the art and means a specific protein molecule directed against an antigenic site.
  • the antibody means an antibody that specifically binds to the protein of the present invention, and such an antibody can be produced by a conventional method by cloning each gene into an expression vector according to a conventional method to obtain a protein encoded by the marker gene, and from the obtained protein.
  • This also includes a partial peptide that can be produced from the protein, and the partial peptide of the present invention includes at least 7 amino acids, preferably 9 amino acids, and more preferably 12 or more amino acids.
  • the form of the antibody of the present invention is not particularly limited, and a polyclonal antibody, a monoclonal antibody, or a part thereof having antigen binding properties are also included in the antibody of the present invention, and all immunoglobulin antibodies are included. Furthermore, the antibody of the present invention also includes special antibodies such as humanized antibodies.
  • the antibody of the present invention includes a complete form having two full-length light chains and two full-length heavy chains, as well as a functional fragment of the antibody molecule.
  • a functional fragment of an antibody molecule means a fragment that possesses at least an antigen-binding function, such as Fab, F(ab'), F(ab') 2, and Fv.
  • the antibody specifically binds to the DKK1 protein or an antigenic determinant thereof expressed on the surface of regulatory T cells to inhibit the activity of regulatory T cells, thereby effectively preventing, improving or treating immune-related diseases, such as fibrosis or cancer.
  • the cancer may be a solid tumor formed by a mass of cells growing abnormally in a solid organ, and depending on the location of the solid organ, may be gastric cancer, liver cancer, glioma, ovarian cancer, colon cancer, head and neck cancer, bladder cancer, renal cell cancer, breast cancer, metastatic cancer, prostate cancer, pancreatic cancer, melanoma, or lung cancer, and preferably, melanoma, but is not limited thereto.
  • the fibrosis may be selected from the group consisting of fibrosis occurring in at least one organ selected from the group consisting of kidney, liver, lung, heart, bone or bone marrow, and skin, but is not limited thereto.
  • the fibrosis includes, but is not limited to, pulmonary fibrosis, liver fibrosis, cardiac fibrosis, brain fibrosis, skin fibrosis, pancreatic fibrosis, and bone fibrosis.
  • fibrosis refers to the development of excessive fibrous connective tissue within an organ or tissue. Fibrosis is characterized by the accumulation and remodeling of the extracellular matrix (ECM). Despite the clear pathognomonic and clinical differences that distinguish it from cancer, which is a mass formed by abnormal cell growth, most chronic fibrotic disorders share in common a persistent stimulus that sustains the production of growth factors, proteolytic enzymes, angiogenic factors, and fibrogenic cytokines, which together stimulate the deposition of connective tissue elements, particularly collagen and proteoglycans, thereby continually remodeling and destroying normal tissue architecture.
  • ECM extracellular matrix
  • the antibody can effectively prevent, improve or treat immune-related diseases, such as fibrosis or cancer, by specifically binding to the DKK-1 protein or an antigenic determinant thereof expressed on the surface of regulatory T cells and increasing the activity of regulatory T cells.
  • a pharmaceutical composition for preventing or treating fibrosis or cancer comprising as an active ingredient any one selected from the group consisting of antisense oligonucleotides, siRNA, shRNA and microRNA specific for the DKK-1 extracellular surface protein of the present invention or a gene encoding the antigenic determinant.
  • the "antisense oligonucleotide” is a short length of DNA synthetic strand (or DNA analog) that is antisense (or complementary) to a specific DNA or RNA target, and is used to achieve gene-specific inhibition in vitro as well as in vivo.
  • Antisense oligonucleotides have been proposed to block the expression of a protein encoded by a DNA or RNA target by binding to the target and stopping expression at the stage of transcription, translation or splicing. Antisense oligonucleotides have also been successfully used in cell culture and animal models of diseases.
  • antisense oligonucleotides to make the oligonucleotides more stable and resistant to degradation are known and understood by those skilled in the art.
  • the antisense oligonucleotides used herein include double-stranded or single-stranded DNA, double-stranded or single-stranded RNA, DNA/RNA hybrids, DNA and RNA analogs, and oligonucleotides having base, sugar or backbone modifications.
  • the oligonucleotides are modified by methods known in the art to increase stability and increase resistance to nuclease degradation. These modifications include, but are not limited to, modifications of the oligonucleotide backbone, modifications of the sugar moieties, or modifications of the bases, which are known in the art.
  • the "siRNA (small interfering RNA)" is a nucleic acid molecule capable of mediating RNA interference or gene silencing, and is used as an efficient gene knockdown method or gene therapy method because it can suppress the expression of a target gene.
  • siRNA molecule When an siRNA molecule is used in the present invention, it may have a structure in which a sense strand (a sequence corresponding to a WLS mRNA sequence) and an antisense strand (a sequence complementary to a WLS mRNA sequence) are positioned on opposite sides to form a double-stranded structure, or a single-stranded structure having self-complementary sense and antisense strands.
  • siRNA is not limited to a double-stranded RNA portion in which RNAs are paired completely, and may include a portion that does not pair due to a mismatch (corresponding bases are not complementary), a bulge/protrusion (there is no base corresponding to one chain), etc.
  • the siRNA terminal structure can be either a blunt end or a sticky end as long as it can suppress the expression of the WLS gene by the RNA interference (RNAi) effect.
  • the sticky end structure can be either a 3'-terminal overhang structure or a 5'-terminal overhang structure.
  • the siRNA molecule may have a total length of 15 to 30 bases, preferably 19 to 21 bases, but is not limited thereto.
  • the "shRNA (short hairpin RNA)" is a single-stranded RNA having a length of 45 to 70 nucleotides, and when oligo DNA is synthesized to connect a 3 to 10 nucleotide linker between the sense strand of the target gene siRNA base sequence and the complementary antisense strand, and then cloned into a plasmid vector or the shRNA is expressed by inserting it into a retrovirus such as a lentivirus and an adenovirus, an shRNA having a hairpin structure with a loop is created and converted into siRNA by dicer within the cell to exhibit an RNAi effect.
  • a retrovirus such as a lentivirus and an adenovirus
  • microRNA regulates various biological processes such as development, differentiation, proliferation, preservation, and apoptosis.
  • MicroRNA generally regulates the expression of a gene encoding a target mRNA by destabilizing the target mRNA or interfering with its translation.
  • useful regulatory sequences e.g., constitutive promoter, inducible promoter, tissue-specific promoter or combinations thereof
  • useful regulatory sequences for the expression construct/vector containing the antisense oligonucleotide, siRNA, shRNA or microRNA can also be appropriately selected from those known in the art.
  • the antisense oligonucleotide, siRNA, shRNA or microRNA can specifically bind to a gene encoding DKK1 protein or an antigenic determinant thereof expressed on the surface of regulatory T cells and inhibit their expression, thereby effectively preventing, improving or treating immune-related diseases, such as cancer.
  • the cancer may be a solid tumor formed by a mass of cells growing abnormally in a solid organ, and depending on the location of the solid organ, may be gastric cancer, liver cancer, glioma, ovarian cancer, colon cancer, head and neck cancer, bladder cancer, renal cell cancer, breast cancer, metastatic cancer, prostate cancer, pancreatic cancer, melanoma, or lung cancer, and preferably, melanoma, but is not limited thereto.
  • the antisense oligonucleotide, siRNA, shRNA or microRNA can effectively prevent, improve or treat immune-related diseases, such as fibrosis or cancer, by specifically binding to a gene encoding DKK1 protein or an antigenic determinant thereof expressed on the surface of regulatory T cells and increasing their expression.
  • prevention may include, without limitation, any act of blocking, suppressing or delaying symptoms of a disease by using the pharmaceutical composition of the present invention.
  • treatment may include without limitation any act that improves or benefits the symptoms of a disease by using the pharmaceutical composition of the present invention.
  • the pharmaceutical composition may be characterized as being in the form of a capsule, tablet, granule, injection, ointment, powder or beverage, and the pharmaceutical composition may be characterized as being intended for humans.
  • the pharmaceutical composition according to the present invention is prepared in the form of an injection and can be directly injected into a site where cancer or an immune disease has occurred, but is not limited thereto.
  • the pharmaceutical composition of the present invention is not limited thereto, but may be formulated and used in the form of oral dosage forms such as powders, granules, capsules, tablets, and aqueous suspensions, external preparations, suppositories, and sterile injection solutions, respectively, according to conventional methods.
  • the pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier.
  • Pharmaceutically acceptable carriers may include binders, lubricants, disintegrants, excipients, solubilizers, dispersants, stabilizers, suspending agents, pigments, fragrances, etc. for oral administration, and may include buffers, preservatives, analgesics, solubilizers, isotonic agents, stabilizers, and the like for injections.
  • compositions of the present invention may be prepared in various ways by mixing with the pharmaceutically acceptable carriers described above.
  • pharmaceutically acceptable carriers for oral administration, it can be manufactured in the form of tablets, troches, capsules, elixirs, suspensions, syrups, wafers, etc., and for injections, it can be manufactured in the form of unit dose ampoules or multiple doses.
  • it can be formulated in the form of solutions, suspensions, tablets, capsules, sustained-release preparations, etc.
  • examples of carriers, excipients and diluents suitable for formulation include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate or mineral oil.
  • fillers, anticoagulants, lubricants, wetting agents, fragrances, emulsifiers and preservatives may be additionally included.
  • the routes of administration of the pharmaceutical composition according to the present invention include, but are not limited to, oral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, enteral, topical, sublingual, or rectal. Oral or parenteral administration is preferred.
  • parenteral as used herein includes subcutaneous, intradermal, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intrathecal, intralesional, and intracranial injection or infusion techniques.
  • the pharmaceutical composition of the present invention may also be administered in the form of a suppository for rectal administration.
  • the pharmaceutical composition of the present invention may vary depending on various factors including the activity of the specific compound used, age, body weight, general health, sex, dosage form, administration time, administration route, excretion rate, drug combination, and the severity of the specific disease to be prevented or treated, and the dosage of the pharmaceutical composition may vary depending on the patient's condition, body weight, degree of disease, drug form, administration route, and period, but may be appropriately selected by those skilled in the art, and may be administered at 0.0001 to 50 mg/kg or 0.001 to 50 mg/kg per day. The administration may be administered once a day or divided into several times. The dosage does not limit the scope of the present invention in any way.
  • the pharmaceutical composition according to the present invention may be formulated as a pill, a dragee, a capsule, a solution, a gel, a syrup, a slurry, or a suspension.
  • the pharmaceutical composition of the present invention can additionally be administered in combination with other anticancer agents, thereby effectively inhibiting general cancer cell proliferation and cancer metastasis, and can thus be used for the treatment of cancer.
  • the above anticancer agents include nitrogen mustard, imatinib, oxaliplatin, rituximab, erlotinib, neratinib, lapatinib, gefitinib, vandetanib, nilotinib, semasanib, bosutinib, axitinib, cediranib, lestaurtinib, trastuzumab, gefitinib, bortezomib, sunitinib, carboplatin, sorafenib, bevacizumab, cisplatin, cetuximab, viscum album, asparaginase, tretinoin, hydroxycarbamide, dasatinib, estramustine, gemtuzumab ozogamicin, ibritumomab tycetan, heptaplatin, methylaminolevulinic acid, amsacrine, alemtuzum
  • the pharmaceutical composition of the present invention can additionally be administered in combination with other immunosuppressants, thereby effectively suppressing diseases caused by general hyperimmune reactions.
  • the immunosuppressant may be at least one selected from the group consisting of glucocorticoids, cyclophosphamide, cyclosporin, tacrolimus, rapamycin, type IV PDE inhibitors, p38 kinase inhibitors, azathioprine, mycophenolate mofetil, mizoribin, methotrexate, leflunomide, and brequina, but is not limited thereto.
  • a method for preventing or treating an immune-related disease comprising administering to a subject in need of treatment an antibody specific for the DKK-1 extracellular surface protein of the present invention or an antigenic determinant.
  • the subject requiring the treatment is an individual having an immune-related disease or suspected of having symptoms thereof
  • the cancer may be gastric cancer, liver cancer, glioma, ovarian cancer, colon cancer, head and neck cancer, bladder cancer, renal cell carcinoma, breast cancer, metastatic cancer, prostate cancer, pancreatic cancer, melanoma, or lung cancer
  • the fibrosis may be selected from the group consisting of, but not limited to, fibrosis occurring in at least one organ selected from the group consisting of the kidney, liver, lung, heart, bone or bone marrow, and skin.
  • the antibody specifically binds to the DKK-1 protein or an antigenic determinant thereof expressed on the surface of regulatory T cells to inhibit the activity of regulatory T cells, thereby effectively preventing, improving or treating immune-related diseases, such as fibrosis or cancer.
  • a method for preventing or treating an immune-related disease comprising administering to a subject in need of treatment any one selected from the group consisting of antisense oligonucleotides, siRNAs, shRNAs and microRNAs specific for the DKK-1 extracellular surface protein of the present invention or a gene encoding the antigenic determinant.
  • the subject requiring the treatment is an individual having an immune-related disease or suspected of having symptoms thereof
  • the cancer may be gastric cancer, liver cancer, glioma, ovarian cancer, colon cancer, head and neck cancer, bladder cancer, renal cell carcinoma, breast cancer, metastatic cancer, prostate cancer, pancreatic cancer, melanoma, or lung cancer
  • the fibrosis may be selected from the group consisting of, but not limited to, fibrosis occurring in at least one organ selected from the group consisting of the kidney, liver, lung, heart, bone or bone marrow, and skin.
  • the antisense oligonucleotide, siRNA, shRNA or microRNA can specifically bind to a gene encoding DKK1 protein or an antigenic determinant thereof expressed on the surface of regulatory T cells and inhibit their expression, thereby effectively preventing, improving or treating immune-related diseases, such as cancer.
  • the antisense oligonucleotide, siRNA, shRNA or microRNA can effectively prevent or treat an immune-related disease by specifically binding to a gene encoding DKK1 protein or an antigenic determinant thereof expressed on the surface of regulatory T cells and increasing their expression.
  • an example of the immune-related disease may be cancer such as gastric cancer, liver cancer, glioma, ovarian cancer, colon cancer, head and neck cancer, bladder cancer, renal cell carcinoma, breast cancer, metastatic cancer, prostate cancer, pancreatic cancer, melanoma or lung cancer, and as another example, the fibrosis may be selected from the group consisting of fibrosis occurring in at least one organ selected from the group consisting of kidney, liver, lung, heart, bone or bone marrow and skin, but is not limited thereto.
  • cancer such as gastric cancer, liver cancer, glioma, ovarian cancer, colon cancer, head and neck cancer, bladder cancer, renal cell carcinoma, breast cancer, metastatic cancer, prostate cancer, pancreatic cancer, melanoma or lung cancer
  • the fibrosis may be selected from the group consisting of fibrosis occurring in at least one organ selected from the group consisting of kidney, liver, lung, heart, bone or bone marrow and skin, but is not limited thereto.
  • administration means introducing the composition of the present invention into a patient by any appropriate method, and the route of administration of the composition of the present invention may be administered through any general route as long as it can reach the target tissue.
  • Oral administration, intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, intranasal administration, intrapulmonary administration, rectal administration, intracavity administration, intraperitoneal administration, and intrathecal administration may be performed, but are not limited thereto.
  • the treatment method of the present invention may include administering the antibody, antisense oligonucleotide, siRNA, shRNA, or microRNA in a pharmaceutically effective amount.
  • the effective amount may be adjusted according to various factors including the type of disease, the severity of the disease, the type and content of the effective ingredient and other ingredients contained in the composition, the type of formulation, and the patient's age, weight, general health, sex, and diet, administration time, administration route, and secretion rate of the composition, treatment period, and concurrently used drugs.
  • the expression inhibitor of the gene or the activity inhibitor of the protein may be administered once or several times a day at a dose of 0.01 ng/kg-10 mg/kg for siRNA, 0.01 ng/kg-10 mg/kg for antisense oligonucleotide to mRNA of the gene, 0.1 ng/kg-10 mg/kg for compound, and 0.1 ng/kg-10 mg/kg for monoclonal antibody to the protein.
  • additional administration may be performed in combination with another anticancer agent or another immunosuppressant.
  • composition for diagnosing fibrosis or cancer comprising an antibody specific for the DKK1 extracellular surface protein or the antigenic determinant of the present invention as an active ingredient.
  • diagnosis means confirming the presence or characteristics of a pathological condition, and for the purpose of the present invention, diagnosis is confirming the presence or absence of gastric cancer and metastasis.
  • the DKK1 extracellular surface protein may be present on the surface of regulatory T cells.
  • fibrosis or cancer can be diagnosed by measuring the expression level of DKK1 extracellular surface protein or the antigen determinant present on the surface of regulatory T cells using the antibody.
  • the immune-related disease may be cancer, and more specifically, may be cancer, liver cancer, glioma, ovarian cancer, colon cancer, head and neck cancer, bladder cancer, renal cell carcinoma, breast cancer, metastatic cancer, prostate cancer, pancreatic cancer, melanoma or lung cancer, and preferably may be melanoma, but is not limited thereto.
  • the fibrosis may be selected from the group consisting of fibrosis occurring in at least one organ selected from the group consisting of kidney, liver, lung, heart, bone or bone marrow and skin, but is not limited thereto.
  • composition for diagnosing fibrosis or cancer comprising any one selected from the group consisting of a primer, a probe and an antisense nucleotide specific for the DKK1 extracellular surface protein of the present invention or a gene encoding the antigenic determinant.
  • the DKK1 extracellular surface protein may be present on the surface of regulatory T cells.
  • fibrosis or cancer can be diagnosed by measuring the expression level of the DKK1 extracellular surface protein present on the surface of regulatory T cells or the gene encoding the antigenic determinant using the antibody.
  • the immune-related disease may be cancer, and more specifically, may be cancer, liver cancer, glioma, ovarian cancer, colon cancer, head and neck cancer, bladder cancer, renal cell carcinoma, breast cancer, metastatic cancer, prostate cancer, pancreatic cancer, melanoma or lung cancer, and preferably may be melanoma, but is not limited thereto.
  • the fibrosis may be selected from the group consisting of fibrosis occurring in at least one organ selected from the group consisting of kidney, liver, lung, heart, bone or bone marrow and skin, but is not limited thereto.
  • a diagnostic kit for fibrosis or cancer comprising the diagnostic composition of the present invention is provided.
  • the kit refers to a set of compositions and accessories necessary for a specific purpose.
  • the kit of the present invention can diagnose the above diseases by confirming the expression level of the DKK1 extracellular surface protein, which is a diagnostic marker of immune-related diseases such as cancer, autoimmune disease, graft-versus-host disease, organ transplant rejection, asthma, atopy, and acute and chronic inflammatory diseases, as mRNA or the expression level of its protein.
  • the kit of the present invention may include a primer, a probe, or optionally an antibody recognizing the marker for measuring the expression level of the diagnostic marker of the immune-related disease, as well as one or more other component compositions, solutions, or devices suitable for the analysis method.
  • a method for providing information on fibrosis or cancer by measuring the expression level of the extracellular surface protein or antigenic determinant of DKK1 present in regulatory T cells using an antibody specific for the extracellular surface protein or the antigenic determinant.
  • the immune-related disease may be cancer, and more specifically, may be cancer, liver cancer, glioma, ovarian cancer, colon cancer, head and neck cancer, bladder cancer, renal cell carcinoma, breast cancer, metastatic cancer, prostate cancer, pancreatic cancer, melanoma or lung cancer, and preferably may be melanoma, but is not limited thereto.
  • the fibrosis may be selected from the group consisting of fibrosis occurring in at least one organ selected from the group consisting of kidney, liver, lung, heart, bone or bone marrow and skin, but is not limited thereto.
  • a method for providing information on fibrosis or cancer by measuring the expression level of a gene encoding the DKK1 extracellular surface protein of the present invention or the antigenic determinant in a regulatory T cell using any one selected from the group consisting of a primer, a probe and an antisense nucleotide specific for the gene.
  • the immune-related disease may be cancer, and more specifically, may be cancer, liver cancer, glioma, ovarian cancer, colon cancer, head and neck cancer, bladder cancer, renal cell carcinoma, breast cancer, metastatic cancer, prostate cancer, pancreatic cancer, melanoma or lung cancer, and preferably may be melanoma, but is not limited thereto.
  • the fibrosis may be selected from the group consisting of fibrosis occurring in at least one organ selected from the group consisting of kidney, liver, lung, heart, bone or bone marrow and skin, but is not limited thereto.
  • the present invention relates to a diagnostic device for fibrosis or cancer.
  • the diagnostic device of the present invention relates to a diagnostic device for fibrosis or cancer, comprising: (a) a measuring unit for measuring the expression level of DKK1 or a gene encoding the protein for a biological sample obtained from a target subject; and (b) a detection unit for outputting the presence or absence of cancer from the expression level of the gene or the protein encoded thereby measured by the measuring unit.
  • the "target subject” refers to a subject that has developed or is likely to develop fibrosis or cancer, and may be a mammal including a human, and may be selected from the group consisting of, for example, a human, a rat, a mouse, a guinea pig, a hamster, a rabbit, a monkey, a dog, a cat, a cow, a horse, a pig, a sheep, and a goat, and is preferably a human, but is not limited thereto.
  • composition for diagnosing fibrosis or cancer comprising an antibody or antigen-binding fragment that specifically binds to an extracellular surface protein of DKK-1 or a DKK-1 epitope represented by any one of SEQ ID NOS: 4 to 36.
  • a diagnostic composition wherein the DKK-1 is present on the surface of regulatory T cells or is present in a dissolved state after being separated from the cells.
  • a diagnostic composition wherein the fibrosis is selected from the group consisting of fibrosis occurring in at least one organ selected from the group consisting of kidney, liver, lung, heart, bone or bone marrow and skin.
  • a diagnostic composition wherein the cancer is a solid cancer.
  • the cancer is at least one selected from the group consisting of gastric cancer, thyroid cancer, parathyroid cancer, ovarian cancer, colon cancer, pancreatic cancer, liver cancer, breast cancer, cervical cancer, lung cancer, non-small cell lung cancer, prostate cancer, gallbladder cancer, biliary tract cancer, non-Hodgkin's lymphoma, Hodgkin's lymphoma, blood cancer, bladder cancer, kidney cancer, melanoma, colon cancer, bone cancer, skin cancer, head cancer, uterine cancer, rectal cancer, brain tumor, anal cancer, fallopian tube carcinoma, endometrial carcinoma, vaginal cancer, vulvar carcinoma, esophageal cancer, small intestine cancer, endocrine cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, ureteral cancer, renal cell carcinoma, renal pelvic carcinoma, central nervous system (CNS) tumor, primary CNS lymphoma, spinal cord tumor, brainstem gliom
  • CNS central nervous system
  • composition for diagnosing fibrosis or cancer comprising a primer, probe or antisense nucleotide specific for a gene encoding an epitope of DKK-1 extracellular surface protein or DKK-1 represented by any one of SEQ ID NOS: 4 to 36.
  • a diagnostic composition wherein the DKK-1 is present on the surface of regulatory T cells or is present in a dissolved state after being separated from the cells.
  • a diagnostic composition wherein the fibrosis is selected from the group consisting of fibrosis occurring in at least one organ selected from the group consisting of kidney, liver, lung, heart, bone or bone marrow and skin.
  • a diagnostic composition wherein the cancer is a solid cancer.
  • the cancer is at least one selected from the group consisting of gastric cancer, thyroid cancer, parathyroid cancer, ovarian cancer, colon cancer, pancreatic cancer, liver cancer, breast cancer, cervical cancer, lung cancer, non-small cell lung cancer, prostate cancer, gallbladder cancer, biliary tract cancer, non-Hodgkin's lymphoma, Hodgkin's lymphoma, blood cancer, bladder cancer, kidney cancer, melanoma, colon cancer, bone cancer, skin cancer, head cancer, uterine cancer, rectal cancer, brain tumor, anal cancer, fallopian tube carcinoma, endometrial carcinoma, vaginal cancer, vulvar carcinoma, esophageal cancer, small intestine cancer, endocrine cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, ureteral cancer, renal cell carcinoma, renal pelvic carcinoma, central nervous system (CNS) tumor, primary CNS lymphoma, spinal cord tumor, brainstem gliom
  • CNS central nervous system
  • a diagnostic kit for fibrosis or cancer comprising the composition is provided.
  • a method for providing information on fibrosis or cancer by measuring the expression level of the extracellular surface protein of DKK-1 or an epitope of DKK-1 represented by any one of SEQ ID NOS: 4 to 36 using an antibody or antigen-binding fragment that specifically binds to the surface protein or epitope.
  • a method for providing information wherein the DKK-1 is present on the surface of a regulatory T cell or is present in a dissolved state after being separated from the cell.
  • a method for providing information on fibrosis or cancer by measuring the expression level of a gene using any one selected from the group consisting of a primer, a probe and an antisense nucleotide specific for a gene encoding an extracellular surface protein of DKK-1, or an epitope of DKK-1 represented by any one of SEQ ID NOS: 4 to 36.
  • a method for providing information wherein the DKK-1 is present on the surface of a regulatory T cell or is present in a dissolved state after being separated from the cell.
  • a method of providing information wherein the fibrosis is selected from the group consisting of fibrosis occurring in at least one organ selected from the group consisting of kidney, liver, lung, heart, bone or bone marrow and skin.
  • a method for providing information is provided, wherein the cancer is a solid cancer.
  • a method for providing information wherein the cancer is at least one selected from the group consisting of gastric cancer, thyroid cancer, parathyroid cancer, ovarian cancer, colon cancer, pancreatic cancer, liver cancer, breast cancer, cervical cancer, lung cancer, non-small cell lung cancer, prostate cancer, gallbladder cancer, biliary tract cancer, non-Hodgkin's lymphoma, Hodgkin's lymphoma, blood cancer, bladder cancer, kidney cancer, melanoma, colon cancer, bone cancer, skin cancer, head cancer, uterine cancer, rectal cancer, brain tumor, anal cancer, fallopian tube carcinoma, endometrial carcinoma, vaginal cancer, vulvar carcinoma, esophageal cancer, small intestine cancer, endocrine cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, ureteral cancer, renal cell carcinoma, renal pelvic carcinoma, central nervous system (CNS) tumor, primary CNS lymphoma,
  • a diagnostic device for fibrosis or cancer comprising: (a) a measuring unit for measuring the expression level of DKK-1 or a gene encoding the protein for a biological sample obtained from a target individual; and (b) a detection unit for outputting the presence or absence of cancer from the expression level of the gene or the protein encoded thereby measured by the measuring unit.
  • a diagnostic device wherein the fibrosis is selected from the group consisting of fibrosis occurring in at least one organ selected from the group consisting of kidney, liver, lung, heart, bone or bone marrow and skin.
  • the cancer is at least one selected from the group consisting of gastric cancer, thyroid cancer, parathyroid cancer, ovarian cancer, colon cancer, pancreatic cancer, liver cancer, breast cancer, cervical cancer, lung cancer, non-small cell lung cancer, prostate cancer, gallbladder cancer, biliary tract cancer, non-Hodgkin's lymphoma, Hodgkin's lymphoma, blood cancer, bladder cancer, kidney cancer, melanoma, colon cancer, bone cancer, skin cancer, head cancer, uterine cancer, rectal cancer, brain tumor, anal cancer, fallopian tube carcinoma, endometrial carcinoma, vaginal cancer, vulvar carcinoma, esophageal cancer, small intestine cancer, endocrine cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, ureteral cancer, renal cell carcinoma, renal pelvic carcinoma, central nervous system (CNS) tumor, primary CNS lymphoma, spinal cord tumor, brainstem gliom
  • CNS central nervous system
  • a pharmaceutical composition for preventing or treating fibrosis or cancer comprising as an active ingredient an antibody or antigen-binding fragment that specifically binds to an extracellular surface protein of DKK-1, or an epitope of DKK-1 represented by any one of SEQ ID NOS: 4 to 36.
  • a pharmaceutical composition wherein the DKK-1 is present on the surface of regulatory T cells or is present in a dissolved state after being separated from the cells.
  • a pharmaceutical composition for preventing or treating fibrosis or cancer comprising a DKK-1 inhibitor is provided.
  • a pharmaceutical composition wherein the DKK-1 inhibitor is selected from the group consisting of NCI8642 (Gallocyanine), BHQ880, DKN-01, RH2-18, Hetero-DS, Cyclized Oligopeptide, and endothelin-1 (ET-1).
  • DKK-1 inhibitor is selected from the group consisting of NCI8642 (Gallocyanine), BHQ880, DKN-01, RH2-18, Hetero-DS, Cyclized Oligopeptide, and endothelin-1 (ET-1).
  • a pharmaceutical composition for preventing or treating fibrosis or cancer comprising as an active ingredient any one selected from the group consisting of an antisense oligonucleotide, siRNA, shRNA and microRNA specific for a gene encoding an extracellular surface protein of DKK-1, or an epitope of DKK-1 represented by any one of SEQ ID NOS: 4 to 36.
  • a pharmaceutical composition wherein the DKK-1 is present on the surface of regulatory T cells or is present in a dissolved state after being separated from the cells.
  • a pharmaceutical composition wherein the fibrosis is selected from the group consisting of fibrosis occurring in at least one organ selected from the group consisting of kidney, liver, lung, heart, bone or bone marrow and skin.
  • a pharmaceutical composition wherein the cancer is a solid cancer.
  • the cancer is at least one selected from the group consisting of gastric cancer, thyroid cancer, parathyroid cancer, ovarian cancer, colon cancer, pancreatic cancer, liver cancer, breast cancer, cervical cancer, lung cancer, non-small cell lung cancer, prostate cancer, gallbladder cancer, biliary tract cancer, non-Hodgkin's lymphoma, Hodgkin's lymphoma, blood cancer, bladder cancer, kidney cancer, melanoma, colon cancer, bone cancer, skin cancer, head cancer, uterine cancer, rectal cancer, brain tumor, anal cancer, fallopian tube carcinoma, endometrial carcinoma, vaginal cancer, vulvar carcinoma, esophageal cancer, small intestine cancer, endocrine cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, ureteral cancer, renal cell carcinoma, renal pelvic carcinoma, central nervous system (CNS) tumor, primary CNS lymphoma, spinal cord tumor, brainstem gliom
  • CNS central nervous system
  • a pharmaceutical composition for preventing or treating fibrosis or cancer comprising as an active ingredient a fusion protein comprising an extracellular surface protein of DKK-1 present in regulatory T cells and an immunoglobulin Fc region.
  • the above fusion protein competitively binds to LRP-5, 6, which are ligands of DKK-1, and ultimately inhibits the function of DKK-1, thereby preventing or treating fibrosis or cancer.
  • Immunoglobulins comprise four polypeptide chains, two heavy chains and two light chains, which are joined by interchain disulfide bonds. Each light chain has two domains, a variable light domain (VL) and a constant light domain (CL), and each heavy chain has two domains, a variable heavy domain (VH) and a constant heavy domain (CH).
  • the constant heavy domain (CH) is made up of constant heavy domains, which are designated by numbers (e.g., CH1, CH2, CH3, etc.). Immunoglobulins are classified into different isotypes (i.e., IgG, IgM, IgA, IgD, and IgE) based on their biological properties, location in an organism, and ability to process different antigens.
  • the constant heavy domain may have three or four CH domains. Additionally, in some isotypes (IgA, IgD, and IgG), the heavy chain contains a hinge region that adds flexibility to the molecule.
  • IgG1 There are four IgG subclasses in humans (IgG1, 2, 3, 4), named according to their order of abundance in serum (IgG1 being the most abundant).
  • An IgG isotype consists of two light chains and two heavy chains, each heavy chain containing three constant heavy domains (CH1, CH2, CH3). The two heavy chains of IgG are linked to each other and to the light chain by disulfide bonds (-S-S-).
  • the antigen-binding site of IgG is located in the fragment antigen-binding region (Fab region), which includes the variable light (VL) and variable heavy (VH) domains, as well as the constant light (CL) and constant heavy (CH1) domains.
  • the fragment crystallizable region (Fc region) of IgG is the part of the heavy chain that contains the CH2 and CH3 domains that bind to Fc receptors found on the surface of certain cells, including the neonatal Fc receptor (FcRn).
  • the heavy chain of IgG also has a hinge region (hinge) between CH1 and CH2 that separates the Fab region from the Fc region and participates in linking the two heavy chains together via a disulfide bond.
  • the structure of the hinge region contributes to the unique biological properties of each of the four IgG subclasses.
  • IgG is secreted as a small monomer that allows it to easily perfuse tissues. It is the only isotype that has a receptor (neonatal Fc receptor (FcRn)) that facilitates its passage across the human placenta to protect the fetus in the womb. IgG absorbed through the placenta provides humoral immunity to the newborn before its own immune system develops.
  • FcRn nonatal Fc receptor
  • the IgG neonatal Fc receptor (FcRn) binding site is located in the Fc region of the antibody.
  • FcRn is commonly expressed on human placental and epithelial cells and participates in the endocytic salvage pathway that protects IgG from degradation. This salvage pathway is mediated by the high pH-dependent binding affinity of IgG for FcRn at acidic pH.
  • the high affinity of IgG for FcRn at acidic pH is thought to drive binding of internalized IgG to FcRn after uptake into acidic endosomes. While most soluble proteins are directed to lysosomes after internalization, internalized FcRn-bound IgG are returned to the plasma membrane and are effectively rescued from the canonical degradation pathway.
  • IgG can dissociate from FcRn and return to the circulation. Thus, the prolonged serum half-life properties of antibodies are maintained in the Fc fragment.
  • the "immunoglobulin Fc region” means a region including a heavy chain constant region 2 (CH2) and/or a heavy chain constant region 3 (CH3) portion, excluding the heavy chain and light chain variable regions of an immunoglobulin.
  • the immunoglobulin Fc region may be a component forming a moiety of the protein complex of the present invention.
  • the immunoglobulin Fc region can affect the structural flexibility of the fusion protein to be finally manufactured, and can further increase the productivity and stability of the fusion protein, but is not limited thereto.
  • the immunoglobulin Fc region may further include a hinge portion in the heavy chain constant region, thereby affecting the structural flexibility of the final fusion protein to be manufactured, and may further increase the productivity and stability of the fusion protein, but is not limited thereto.
  • the immunoglobulin Fc region of the present invention may be an extended Fc region including part or all of the heavy chain constant region 1 (CH1) and/or the light chain constant region 1 (CL1), excluding only the heavy and light chain variable regions of the immunoglobulin, as long as it has substantially the same or improved effect as the native type.
  • it may also be a region in which a relatively long part of the amino acid sequence corresponding to CH2 and/or CH3 is deleted.
  • the immunoglobulin Fc region of the present invention may be a dimer of 1) a CH1 domain, a CH2 domain, a CH3 domain, and a CH4 domain, 2) a CH1 domain and a CH2 domain, 3) a CH1 domain and a CH3 domain, 4) a CH2 domain and a CH3 domain, 5) a combination of one or more of the CH1 domain, the CH2 domain, the CH3 domain, and the CH4 domain with an immunoglobulin hinge region (or a part of the hinge region), 6) each domain of a heavy chain constant region and a light chain constant region.
  • an immunoglobulin hinge region or a part of the hinge region
  • the immunoglobulin Fc region of the present invention includes not only a natural amino acid sequence but also a sequence derivative thereof.
  • An amino acid sequence derivative means a sequence having a different sequence due to deletion, insertion, non-conservative or conservative substitution, or a combination thereof, of one or more amino acid residues in a natural amino acid sequence.
  • amino acid residues 214 to 238, 297 to 299, 318 to 322, or 327 to 331, which are known to be important for binding can be used as suitable sites for modification.
  • various types of derivatives are possible, such as those in which a site capable of forming a disulfide bond is removed, in which several amino acids at the N-terminus of a native Fc are removed, or in which a methionine residue is added to the N-terminus of a native Fc.
  • a complement binding site e.g., a C1q binding site
  • ADCC antibody dependent cell mediated cytotoxicity
  • Amino acid exchanges in proteins and peptides that do not alter the overall activity of the molecule are well known in the art (H.Neurath, R.L.Hill, The Proteins, Academic Press, New York, 1979).
  • the most common exchanges are between amino acid residues Ala/Ser, Val/Ile, Asp/Glu, Thr/Ser, Ala/Gly, Ala/Thr, Ser/Asn, Ala/Val, Ser/Gly, Thy/Phe, Ala/Pro, Lys/Arg, Asp/Asn, Leu/Ile, Leu/Val, Ala/Glu, Asp/Gly.
  • Modifications may also occur, such as phosphorylation, sulfation, acrylation, glycosylation, methylation, farnesylation, acetylation, and amidation.
  • the Fc derivative described above exhibits biological activity equivalent to the Fc region of the present invention and may have enhanced structural stability of the Fc region with respect to heat, pH, etc.
  • the Fc region may be obtained from a natural type isolated from an animal such as a human, cow, goat, pig, mouse, rabbit, hamster, rat or guinea pig, or may be a recombinant or a derivative thereof obtained from a transformed animal cell or microorganism.
  • the method for obtaining from a natural type may be a method of isolating the entire immunoglobulin from a human or animal body and then treating it with a protease to obtain it. When treated with papain, it is cleaved into Fab and Fc, and when treated with pepsin, it is cleaved into pF'c and F(ab)2.
  • the Fc region derived from a human or mouse is a recombinant immunoglobulin Fc region obtained from a microorganism.
  • the immunoglobulin Fc region may be a native sugar chain, a sugar chain with an increased amount compared to the native type, a sugar chain with a decreased amount compared to the native type, or a form in which the sugar chain is removed.
  • Conventional methods such as a chemical method, an enzymatic method, and a genetic engineering method using microorganisms can be used for the increase or removal of the immunoglobulin Fc sugar chain.
  • the immunoglobulin Fc region in which the sugar chain is removed from the Fc has a significantly reduced binding affinity to complement (c1q), and antibody-dependent cytotoxicity or complement-dependent cytotoxicity is reduced or eliminated, so that it does not induce unnecessary immune responses in vivo.
  • the form that is more suitable for the original purpose as a drug carrier is an immunoglobulin Fc region in which the sugar chain is removed or is non-glycosylated.
  • deglycosylation refers to an Fc region from which a sugar has been removed by an enzyme
  • aglycosylation refers to an Fc region that has not been glycosylated and is produced in a prokaryotic animal, or in a more specific embodiment, in E. coli.
  • the immunoglobulin Fc region may be of animal origin, such as human or cow, goat, pig, mouse, rabbit, hamster, rat, or guinea pig, and may be of human or mouse origin as a preferred example.
  • the immunoglobulin Fc region of the present invention may be a hybrid Fc comprising an Fc region derived from IgG, IgA, IgD, IgE, IgM, heavy chain constant region 2 (CH2), heavy chain constant region 3 (CH3), a hinge, a fragment thereof, or a combination thereof, or a combination thereof.
  • a hybrid Fc comprising an Fc region derived from IgG, IgA, IgD, IgE, IgM, heavy chain constant region 2 (CH2), heavy chain constant region 3 (CH3), a hinge, a fragment thereof, or a combination thereof, or a combination thereof.
  • the term "combination” means that when forming a dimer or multimer, a polypeptide encoding a single-chain immunoglobulin Fc region, heavy chain constant region 2 (CH2) or heavy chain constant region 3 (CH3) of the same origin forms a bond with a single-chain polypeptide of different origin. That is, it is possible to produce a dimer or multimer from two or more fragments selected from the Fc region, heavy chain constant region 2 (CH2) or heavy chain constant region 3 (CH3) derived from IgG, IgA, IgM, IgD or IgE.
  • the "hybrid Fc" can be derived from a combination of human IgG subclasses or a combination of human IgD and IgG.
  • the hybrid Fc can include, for example, an IgD hinge region and a CH2 N-terminal region + an IgG4 CH2 and CH3 region, and can be used by borrowing the same hybrid Fc form disclosed in, for example, Korean Patent No. 0897938, which is incorporated herein by reference.
  • the hybrid Fc when the hybrid Fc binds to a biologically active molecule, a polypeptide, etc., it not only increases the serum half-life of the biologically active molecule, but also has the effect of increasing the expression level of the polypeptide when a nucleotide encoding an Fc-polypeptide fusion protein is expressed.
  • the immunoglobulin Fc region may be an Fc region derived from IgG, IgA, IgM, IgD or IgE, or may include a heavy chain constant region 2 (CH2) and a heavy chain constant region 3 (CH3) derived from IgG, IgA, IgM, IgD or IgE, but is not limited thereto.
  • the immunoglobulin Fc region may be an Fc region derived from IgG or IgM which is most abundant in human blood, or may include a heavy chain constant region 2 (CH2) and a heavy chain constant region 3 (CH3) derived from IgG or IgM, or as another example, an Fc region derived from IgG known to enhance the half-life of a ligand binding protein, or may include a heavy chain constant region 2 (CH2) and a heavy chain constant region 3 (CH3) derived from IgG, or as another example, an Fc region derived from IgG1, IgG2, IgG3 or IgG4, or may include a heavy chain constant region 2 (CH2) and a heavy chain constant region 3 (CH3) derived from IgG1 or IgG2.
  • CH2 heavy chain constant region 2
  • CH3 heavy chain constant region 3
  • the immunoglobulin Fc region may include a heavy chain constant region 2 (CH2) and a heavy chain constant region 3 (CH3) derived from human IgG1 represented by SEQ ID NO: 37, or may include a heavy chain constant region 2 (CH2) and a heavy chain constant region 3 (CH3) derived from mouse IgG2 represented by SEQ ID NO: 38, but is not limited thereto.
  • CH2 heavy chain constant region 2
  • CH3 heavy chain constant region 3
  • mouse IgG2 represented by SEQ ID NO: 38
  • the immunoglobulin Fc region may include a hinge region derived from IgG, IgA, IgM, IgD, IgE, or Abatacept, or as another example, may include a hinge region derived from IgG, IgD or Abatacept, or may include a hinge region derived from IgG1, IgG2, IgG3, IgG4, IgD or Abatacept, but is not limited thereto.
  • the immunoglobulin Fc region may include at least one selected from the group consisting of a hinge region derived from human IgG1 represented by SEQ ID NO: 39; a hinge region derived from mouse IgG2 represented by SEQ ID NO: 40; a hinge region derived from human IgD represented by SEQ ID NO: 41; and a hinge region of Abatacept represented by SEQ ID NO: 42; thereby increasing the structural flexibility of the fusion protein to be finally manufactured and significantly improving the productivity and stability of the fusion protein, but is not limited thereto.
  • the linker when the extracellular domain and the Fc region of the DKK-1 protein are connected via a linker, the linker can be connected to the N-terminus, the C-terminus or a free radical of the Fc fragment, and can be connected to the N-terminus, the C-terminus or a free radical of the extracellular domain of the DKK-1 protein.
  • the linker is a peptide linker, the connection can occur at any site.
  • the linker can be connected to the C-terminus of the extracellular domain of the DKK-1 protein and the N-terminus of the Fc region of the immunoglobulin, or can be connected to the C-terminus of the Fc region of the immunoglobulin and the N-terminus of the extracellular domain of the DKK-1 protein.
  • the "linker” can reduce the interference effect between the extracellular domain of the DKK-1 protein and the immunoglobulin Fc region in the fusion protein, thereby increasing the desired activity of the extracellular domain of the DKK-1 protein in the target cell.
  • the linker in the present invention can include a sequence that can be cleaved by an enzyme that is overexpressed in the tissue or cell of the desired disease. In the case where it can be cleaved by the overexpressed enzyme as described above, the activity of the polypeptide can be effectively prevented from being reduced due to the Fc portion.
  • a peptide linker composed of 33 amino acids located at positions 282 to 314 of human albumin, which is most abundant in blood more preferably a peptide linker composed of 13 amino acids located at positions 292 to 304, is a portion that is mostly exposed to the outside in the three-dimensional structure and thus has a minimal possibility of inducing an immune response in the body.
  • it is not limited thereto.
  • the linker when the linker and Fc region in the present invention are expressed separately and then linked to each other, the linker may be a cross-linking agent known in the art.
  • the crosslinking agent may be, but is not limited to, imidoesters including, for example, 1,1-bis(diazoacetyl)-2-phenylethane, glutaraldehyde, N-hydroxysuccinimide esters such as 4-azidosalicylic acid, disuccinimidyl esters such as 3,3'-dithiobis(succinimidylpropionate), and bifunctional maleimides such as bis-N-maleimido-1,8-octane.
  • imidoesters including, for example, 1,1-bis(diazoacetyl)-2-phenylethane, glutaraldehyde, N-hydroxysuccinimide esters such as 4-azidosalicylic acid, disuccinimidyl esters such as 3,3'-d
  • a pharmaceutical composition for preventing or treating fibrosis or cancer comprising a fusion protein comprising LRP-5, 6 and an immunoglobulin Fc region as an active ingredient.
  • the above fusion protein contains LRP-5, 6, which is a ligand that binds to DKK-1, LRP-5, 6 and the above fusion protein competitively bind to DKK-1, and thus fibrosis or cancer can be treated by competitively inhibiting DKK-1.
  • the above fusion protein has been described in detail in the fusion protein comprising DKK-1 and the immunoglobulin Fc region discussed above, and is omitted to avoid duplication.
  • a pharmaceutical composition wherein the DKK-1 is present on the surface of regulatory T cells or is present in a dissolved state after being separated from the cells.
  • Figure 1 shows that DKK-1 expression levels are elevated in patients with human idiopathic pulmonary fibrosis.
  • Figure 2 shows that the expression of DKK-1 was increased in the lungs stimulated with BLM.
  • Figure 3 shows that DKK-1 hypomorphic Dkk-1 d/d mice are protected from BLM-mediated lung injury.
  • FIG. 4 shows that platelets are the main source of DKK-1 histological observations.
  • FIG. 5 shows the expression of alternatively activated macrophage (AAM) markers mediated by DKK-1 in bone marrow-derived macrophages (BMDM).
  • AAM alternatively activated macrophage
  • FIG. 6 shows that deletion of the DKK1 receptor LRP6 protects mice from BLM-induced injury.
  • Figure 7 shows that delayed DKK1 antibody treatment inhibits collagen deposition in a BLM-induced lung injury model.
  • Example 1 It was confirmed that the level of DKK-1 protein expression was increased in lung tissue of human idiopathic pulmonary fibrosis (IPB) patients.
  • a human lung tissue array kit fixed in paraffin blocks was purchased (Pulmonary interstitial fibrosis tissue array, LC561; Biomax). Each lung tissue was deparaffinized with Histoclear and dehydrated. Antigens were removed with sodium citrate buffer (10 mM, pH 6.0). To inhibit endogenous peroxidase activity, the sections were blocked with 3% H2O2. Each section was blocked in 5% BSA, treated with primary antibody (Goat polyclonal anti-m/hDKK1, R&D systems; AF1096), and stained with secondary antibody at a concentration of 5 ug/mL. Antibody color development was performed using a DAB substrate kit (SK-4100; Vector Laboratories), and H&E staining was performed simultaneously to observe the images. The results are shown in Figure 1.
  • mice C57BL/6 mice were treated intranasally with 4 U/kg of BLM dissolved in 40 ⁇ L PBS. Mice aged 8 to 10 weeks were treated with BLM, and the mice were sacrificed 2 weeks later and their lungs were removed. The removed lungs were fixed in 10% formalin for at least 48 hours after lung perfusion and embedded in paraffin blocks for histological analysis. These were sectioned using a microtome. The paraffin-blocked lung tissues were subjected to immunostaining in the same manner as described in [Example 1]. The results are shown in Fig. 2.
  • BLM-induced lung fibrosis mice group was anesthetized with 1% sodium pentobarbital and administered 4 U/kg BLM (BLM, S121415; Selleckchem, Houston, TX) in 40 ⁇ l saline via the intratracheal route as reported.
  • Mice administered the same amount of saline were used as controls, and mice were sacrificed 14 days after BLM administration for lung fibrosis analysis.
  • Lungs were removed from mice, fixed in 10% formalin for at least 48 h after lung perfusion, and embedded in paraffin blocks for histological analysis. These were sectioned with a microtome. The paraffin-blocked lung tissues were subjected to immunostaining in the same manner as described in [Example 1]. The results are shown in Fig. 3.
  • Platelets were isolated from mice as previously described with some modifications. Blood was collected by cardiac puncture into 3.8% sodium citrate, pH 7.4 (2:1, by volume) and centrifuged at 250 g for 10 min to obtain platelet-rich plasma (PRP). For platelet stimulation, PRP was treated with 100 ⁇ M thrombin receptor activating peptide (SFLLRN; Sigma-Aldrich) for the indicated times and then centrifuged at 657 g for 7 min. The supernatant was collected as plasma, and the platelet pellet was also collected. For platelet release, PRP was centrifuged at 657 g for 7 min to pellet platelets.
  • SFLLRN thrombin receptor activating peptide
  • Platelets were then resuspended in Tyrode-HEPES (N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid) buffer and stimulated with 0.1 U/mL thrombin (Sigma-Aldrich) for 90 min. Platelet release was then collected by centrifugation at 2500 g for 10 min at 4°C.
  • Tyrode-HEPES N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid
  • thrombin Sigma-Aldrich
  • BMDM bone marrow-derived macrophages
  • BMDM differentiation mice were sacrificed, bone marrow cells were extracted on day 0, and cultured in DMEM medium containing 20% FBS, GlutaMAX, MEM-NEAA, sodium pyruvate, 1x penicillin & streptomycin, and 20 ng/mL MCSF (macrophage colony-stimulating factor). The medium was changed every two days, and IL-4 and IL-13 were treated on day 6 to induce differentiation into M2-like macrophages.
  • Example 6 Deletion of the DKK1 receptor LRP6 protects mice from bone marrow-derived macrophage (BMDM)-induced damage.
  • BMDM bone marrow-derived macrophage
  • LRP6 fl/fl and LysM-Cre LRP6 fl/fl conditional KO mice were induced with BLM, and the degree of lung damage was compared through collagen mRNA and immunohistochemical staining.
  • deletion of the DKK1 receptor LRP6 was found to protect mice from BLM-induced damage.
  • Example 7 Delayed DKK1 antibody treatment was confirmed to inhibit collagen deposition in a BLM-induced lung injury model.
  • C57BL6 mice were treated with BLM on day 0, and 150 ⁇ g/mouse DKK-1 antibody was introduced in two administration methods to confirm the therapeutic effect of DKK-1 antibody.
  • the first administration group was treated with antibody on days -1, 1, 3, 7, 9, and 11, and the second was treated on days 7, 9, and 11.
  • both administration groups reduced the expression level of DKK-1, thereby inhibiting collagen deposition in the lungs.
  • Delayed DKK1 antibody treatment also showed sufficient therapeutic effects, inhibiting collagen deposition to a similar degree as the pre-administered group.
  • DKK-1 hypomorphic doubleridge mice reduction in DKK-1 expression protects against BLM-induced inflammation and fibrosis.
  • platelet-specific deletion of DKK-1 in DKK-1 PKO mice reduces AAM markers such as Arg1 + and CD206 + , and DKK-1 induces several proinflammatory and profibrotic markers in the absence of type 2 cytokines.
  • DKK-1 enhances IL-13-mediated AAM phenotype, and co-inhibition of STAT6 and JNK is sufficient to eliminate excessive AAM-like macrophage gene expression.
  • deletion of the DKK-1 receptor in myeloid lineage cells of Lrp6 MKO mice protects against BLM-induced inflammation and fibrosis. Delayed treatment with DKK-1 antibodies suppresses BLM-induced inflammation and fibrosis.
  • DKK-1 is an important target protein that induces the occurrence of fibrosis
  • small molecule substances or monoclonal antibodies that inhibit the function or activity of DKK-1 may be new therapeutic agents for the treatment of fibrosis
  • DKK-1 + Treg cells are important for the control of fibrosis
  • DKK-1 + Treg cells can be used as cell therapy agents.
  • DKK-1 plays an important role in solid cancer mediated by fibrosis
  • inhibition of the function of DKK-1 or DKK-1 + Treg cells or TAM induced by DKK-1 may play an important role in the prevention and treatment of these fibrosis-associated solid cancers.

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Abstract

La présente invention concerne une protéine Dickkopf-1 (DKK1) qui est présente de manière spécifique sur la surface de cellules immunitaires, en particulier de lymphocytes T régulateurs (lymphocytes Treg), un anticorps spécifique à celle-ci, et des utilisations associées. Dans la présente invention, étant donné que la protéine DKK1 est présente de manière spécifique sur la surface de cellules immunitaires, en particulier de lymphocytes T régulateurs, la protéine DKK1 peut être utilisée en tant que nouvelle cible pour des agents thérapeutiques pour diverses maladies liées au système immunitaire.
PCT/KR2024/001828 2023-02-07 2024-02-07 Protéine dkk-1 présente de manière spécifique sur des lymphocytes t régulateurs et son utilisation Ceased WO2024167312A1 (fr)

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Citations (5)

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JP2006217844A (ja) * 2005-02-09 2006-08-24 Univ Of Tokyo Ｄｋｋ１の遺伝子、蛋白質及び抗体を用いた癌の診断・モニター方法及び治療方法
US9751923B2 (en) * 2012-11-01 2017-09-05 University Of Washington Through Its Center For Commercialization Administration of DKK1 muteins to treat fibrosis
US10001484B2 (en) * 2014-03-18 2018-06-19 Sanford Health Reagents and methods for breast cancer detection
KR20180123989A (ko) * 2017-05-10 2018-11-20 주식회사 굳티셀 조절 t 세포에 특이적으로 존재하는 dkk1 단백질 및 그 용도
KR20220131223A (ko) * 2019-11-22 2022-09-27 리프 테라퓨틱스 인코포레이티드 Dkk-1 억제제를 사용하여 암을 치료하는 방법

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JP2006217844A (ja) * 2005-02-09 2006-08-24 Univ Of Tokyo Ｄｋｋ１の遺伝子、蛋白質及び抗体を用いた癌の診断・モニター方法及び治療方法
US9751923B2 (en) * 2012-11-01 2017-09-05 University Of Washington Through Its Center For Commercialization Administration of DKK1 muteins to treat fibrosis
US10001484B2 (en) * 2014-03-18 2018-06-19 Sanford Health Reagents and methods for breast cancer detection
KR20180123989A (ko) * 2017-05-10 2018-11-20 주식회사 굳티셀 조절 t 세포에 특이적으로 존재하는 dkk1 단백질 및 그 용도
KR20220131223A (ko) * 2019-11-22 2022-09-27 리프 테라퓨틱스 인코포레이티드 Dkk-1 억제제를 사용하여 암을 치료하는 방법

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