KR20230060794A - Composition for inhibiting plasticity of cancer initiating cells, comprising an inhibitor of the activity or expression of JAGGED1 - Google Patents

Abstract

The composition for inhibiting cancer-initiating cell reprogramming containing the Jagged1 protein inhibitor of the present invention inhibits the expression and activity of Jag1, an inducer of pancreatic cancer cell plasticity, thereby inhibiting cancer-initiating cells involved in tumor growth, induction of drug resistance, or recurrence of cancer. Since it can reduce plasticity, it can be used as a cancer-initiating cell-targeted therapy for pancreatic cancer.

Description

Composition for inhibiting plasticity of cancer initiating cells, comprising an inhibitor of the activity or expression of JAGGED1

The present invention relates to a composition comprising a JAGGED1 protein activity or expression inhibitor for inhibiting the plasticity of cancer initiating cells and its use.

Pancreatic cancer is one of the malignant tumors with an acute onset, late diagnosis, and poor prognosis with a 5-year survival rate of less than 10%. Pancreatic ductal adenocarcinoma (PDAC) is generally referred to as pancreatic ductal adenocarcinoma (PDAC).

The combination of gemcitabine and nab-paclitaxel or systemic chemotherapy such as FOLFIRINOX increased the survival rate compared to gemcitabine. Although chemotherapy is advancing, cancer recurrence is caused by drug resistance, and many studies have reported that tumor heterogeneity causes drug resistance.

In particular, cancer initiating cells (CICs) have the ability to self-renew and differentiate to reproduce the cell heterogeneity of the original tumor. CIC targeting through inhibition of EPCAM, Hedgehog, Notch/DLL4, Wnt and TGFb signaling showed improved therapeutic efficacy in xenograft models, but clinical application was very limited due to toxicity and low efficacy. The reason for this is the ability of cancer cells to switch between CSCs and non-CSCs controlled by internal and external influences, i.e., the plasticity of cancer cells. Therefore, for a perfect response to anticancer drugs, the development of cancer cell plasticity-targeting therapies is essential.

JAGGED1 (JAG1) is one of the Notch ligands, and its expression is increased in various cancer cells and tissues, and it is known to play an important role in cancer malignancy.

As a prior art related to the role of JAG1 in cancer, Korean Patent Publication No. 10-2014-0131827, “Use of APE/Rvf-1 and JAG1/Ntch as a diagnostic marker for colorectal cancer,” shows that Jagged1 and Notch3 proteins are higher than normal tissues. It is disclosed that the expression level is increased in tissues or cells of colorectal cancer, and thus the proteins can be used as markers for diagnosis of colorectal cancer.

However, little is known about the role of JAG1 in cancer cell plasticity. Accordingly, the present inventors completed the present invention by establishing an organoid-derived PDAC assembly containing endothelial cells and autoimmune cells, and confirming that JAG1 plays a role in a dynamic switch for cancer plasticity in the PDAC assembly. did

An object of the present invention is to provide a composition for inhibiting cancer initiating cell differentiation.

Another object of the present invention is to provide a composition for preventing or treating recurrence or metastasis of pancreatic cancer.

Another object of the present invention is to provide a screening method for a therapeutic agent for recurrence or metastasis of pancreatic cancer.

Another object of the present invention is to provide a composition for diagnosing recurrence or metastasis of pancreatic cancer.

Another object of the present invention is to provide a method for providing information for diagnosing recurrence or metastasis of pancreatic cancer.

However, the technical problem to be achieved by the present invention is not limited to the above-mentioned problems, and other problems not mentioned will be clearly understood by those skilled in the art from the following description.

Hereinafter, various embodiments described herein are described with reference to the drawings. In the following description, numerous specific details are set forth, such as specific forms, compositions, and processes, etc., in order to provide a thorough understanding of the present invention. However, certain embodiments may be practiced without one or more of these specific details, or with other known methods and forms. In other instances, well known processes and manufacturing techniques have not been described in specific detail in order not to unnecessarily obscure the present invention. Reference throughout this specification to “one embodiment” or “an embodiment” means that a particular feature, form, composition or characteristic described in connection with the embodiment is included in one or more embodiments of the invention. Thus, the appearances of "in one embodiment" or "an embodiment" in various places throughout this specification do not necessarily refer to the same embodiment of the invention. Additionally, particular features, forms, compositions, or properties may be combined in one or more embodiments in any suitable way.

Unless otherwise defined in the specification, all scientific and technical terms used herein have the same meaning as commonly understood by a person skilled in the art to which the present invention belongs.

The "cancer initiating cells (CICs)" of the present invention are also defined as cancer stem cells (CSCs), and have self-renewal and differentiation capabilities to reproduce the cell heterogeneity of the original tumor. Cancer-initiating cells play an important role in tumor progression, drug resistance, and relapse of cancer. In particular, cancer cells or tissues of patients with pancreatic ductal adenocarcinoma (PDAC) are cancer initiating cells with CD44(+)CD24(+) and CD44(+)CD24(+)EpCAM(+) phenotypes. cells), cancer cells of the PDAC patients are mostly metastatic, and are highly resistant to classical therapies such as chemotherapy, radiation therapy, and immunotherapy. .

"Jag1 (Jagged1)" is one of five cell surface proteins that interact with four receptors in the Notch signaling pathway, and activation of Notch signaling by intercellular signaling in normal tissues and cancers leads to specific cell fate and cell growth. It plays an important role in acquisition. Notch ligand members are generally distinguished by the presence of N-terminal DSL (Delta/Serrate/LAG-2) motifs and specialized tandem EGF repeats called DOS (Delta and OSM-11 like protein) domains. As canonical Notch ligand members, Dll1, Jag1 and Jag2 have DSL ligands with DOS domains, whereas Dll3, Dll4 have DSL ligands without DOS domains.

Jag1/Notch signaling also plays an important role in the generation and maintenance of cancer initiating cells. The Jag1 protein gene or protein expression or activity inhibitor inhibits cancer-initiating cells and reduces cancer cell plasticity, and thus can be used for prevention or treatment of cancer recurrence or metastasis.

Here, "plasticity" refers to the ability of a cell to reversibly change its phenotype, including both differentiation and dedifferentiation, regardless of class, and in both directions. One example is that epithelial cells acquire mesenchymal characteristics (loss of cell-cell gap, migratory and penetrating ability, etc.) during the early embryonic development of metazoans. called Transition). Through EMT-induced changes in cell adhesion and behavior, cells participate in intra-embryonic migration, long-distance migration, and formation of internal organs. Internal organ formation refers to the reverse process of EMT, in which cells undergo mesenchymal-epithelial transition (MET), in which cells reach their destination and then reverse to an epithelial phenotype, settle, proliferate, and differentiate into individual organs. EMT-induced phenotypic changes are accompanied by dramatic changes in cell behavior, and EMT programs are induced by responses to extracellular signals by activation of transcription factors (EMT-TFs) such as Snail, Twist, and Zeb. EMT-TFs also influence cellular processes that help maintain the mesenchymal phenotype, increase migration efficiency, and survive in adverse environments. Transition of cells from carcinoma to migratory and invasive mesenchymal cells is also an EMT process, and several EMT-TFs have been identified to activate during tumor progression, especially during dissemination of cells from primary tumors. there is a lot of evidence Although developmental and pathological EMTs share many common features, they also have significant differences, so EMT can be classified as type 1-embryonic EMTs; Type 2 - Wound repair, tissue regeneration and organ fibrosis; It is classified as type 3-cancer-associated EMT. During cancer progression, cancer cells, in addition to their ability to invade and migrate, invade and migrate into the lymph and blood vessels of the exfoliated cells, followed by extravasation.

An agent that reduces the activity or expression of the protein may be a compound, a peptide mimetics, an aptamer, an antibody, or a natural product that specifically binds to the Jag1 protein or a part thereof.

The "antibody" refers to a substance that specifically binds to an antigen and causes an antigen-antibody reaction. For the purposes of the present invention, antibody means an antibody that specifically binds to said protein. Antibodies of the present invention include both polyclonal antibodies, monoclonal antibodies and recombinant antibodies. Such antibodies can be readily prepared using techniques well known in the art. For example, polyclonal antibodies can be produced by a method well known in the art, including a process of injecting an antigen of the protein into an animal and collecting blood from the animal to obtain serum containing the antibody. Such polyclonal antibodies can be prepared from any animal, such as goat, rabbit, sheep, monkey, horse, pig, cow, dog, and the like. In addition, monoclonal antibodies can be prepared using the hybridoma method well known in the art (see Kohler and Milstein (1976) European Journal of Immunology 6:511-519), or the phage antibody library technique (Clackson et al, Nature, 352). :624-628, 1991; Marks et al, J. Mol. Biol., 222:58, 1-597, 1991). Antibodies prepared by the above methods may be separated and purified using methods such as gel electrophoresis, dialysis, salt precipitation, ion exchange chromatography, and affinity chromatography. In addition, the antibodies of the present invention include functional fragments of antibody molecules as well as complete forms having two full-length light chains and two full-length heavy chains. A functional fragment of an antibody molecule means a fragment having at least an antigen-binding function, and includes Fab, F(ab'), F(ab')2, and Fv.

The "oligopeptide" is a peptide composed of 2 to 20 amino acids and may include a dipeptide, tripeptide, tetrapeptide and pentapeptide, but is not limited thereto.

The "aptamer" is an oligonucleic acid or peptide molecule, and a general description of aptamers is described in Bock LC et al., Nature 355(6360):5646 (1992); Hoppe-Seyler F, Butz K "Peptide aptamers: powerful new tools for molecular medicine". J Mol Med. 78(8):42630 (2000); Cohen BA, Colas P, Brent R. "An artificial cell-cycle inhibitor isolated from a combinatorial library". Proc Natl Acad Sci USA. 95(24): 142727 (1998).

-턴 디펩티드 코어 (Nagai et al. Tetrahedron Lett 26:647, 1985), 케토-메틸렌 슈도펩티드류 (Ewenson et al. J Med chem 29:295, 1986; 및 Ewenson et al. in Peptides: Structure and Function (Proceedings of the 9th AmeriCan Peptide Symposium) Pierce chemiCal co. Rockland, IL, 1985), 아제핀 (Huffman et al. in Peptides: chemistry and Biology, G.R. Marshall ed., EScOM Publisher: Leiden, Netherlands, 1988), 벤조 디아제핀 (Freidinger et al. in Peptides; chemistry and Biology, G.R. Marshall ed., EScOM Publisher: Leiden, Netherlands, 1988),

-아미노알콜 (Gordon et al. Biochem Biophys Res commun 126:419 1985) 및 치환 감마 락탐환 (Garvey et al. in Peptides: chemistry and Biology, G.R. Marshell ed., EScOM Publisher: Leiden, Netherlands, 1988)을 사용하여 생성할 수 있다.The "peptide mimetics" are peptides or non-peptides that inhibit the binding domain of the Jag1 protein leading to inhibition of the activity of Jag1. The major residues of the non-hydrolyzable peptide analogs include

-turn dipeptide core (Nagai et al. Tetrahedron Lett 26:647, 1985), keto-methylene pseudopeptides (Ewenson et al. J Med chem 29:295, 1986; and Ewenson et al. in Peptides: Structure and Function (Proceedings of the 9th AmeriCan Peptide Symposium) Pierce chemiCal co. Rockland, IL, 1985), azepine (Huffman et al. in Peptides: chemistry and Biology, GR Marshall ed., EScOM Publisher: Leiden, Netherlands, 1988), benzo diazepines (Freidinger et al. in Peptides; chemistry and Biology, GR Marshall ed., EScOM Publisher: Leiden, Netherlands, 1988);

-Using amino alcohols (Gordon et al. Biochem Biophys Res commun 126:419 1985) and substituted gamma-lactam rings (Garvey et al. in Peptides: chemistry and Biology, GR Marshell ed., EScOM Publisher: Leiden, Netherlands, 1988) can be created by

An agent that reduces the expression level of the gene may be an antisense nucleotide, small interfering RNA (siRNA), short hairpin RNA (shRNA) or ribozyme that complementarily binds to the Jagged1 gene or a part thereof.

The "antisense nucleotide", as defined in Watson-Crick base pairing, interferes with the flow of genetic information from DNA to protein by binding (hybridizing) to a complementary nucleotide sequence of DNA, immature-mRNA or mature mRNA. The specific nature of antisense nucleotides for target sequences makes them exceptionally multifunctional. Since antisense nucleotides are long chains of monomeric units, they can be easily synthesized against the target RNA sequence. Recently, many studies have demonstrated the usefulness of antisense nucleotides as a biochemical means to study target proteins. Since many recent advances have been made in the field of oligonucleotide chemistry and the synthesis of nucleotides that exhibit improved cell line adsorption, target binding affinity, and nuclease resistance, the use of antisense nucleotides can be considered as a new type of inhibitor.

The "shRNA" and "siRNA" are nucleic acid molecules that can mediate RNA interference or gene silencing, and can suppress the expression of a target gene, so they can be used as an efficient gene knockdown method or gene therapy method do. shRNA is a hairpin structure formed by binding between complementary sequences within a single-stranded oligonucleotide, and in vivo, the shRNA is cleaved by dicer to form small RNA fragments of 21 to 25 nucleotides in size siRNA, which is a double-stranded oligonucleotide, can specifically bind to mRNA with a complementary sequence and suppress its expression. In addition, siRNA refers to a small double-stranded RNA fragment with a size of 21 to 25 nucleotides that induces RNA interference (RNAi) by modifying target mRNA by double-stranded RNA (dsRNA).

Another embodiment of the present invention provides a composition for preventing or treating recurrence or metastasis of pancreatic cancer.

The composition may include an agent that reduces the activity or expression level of the Jag1 protein; Alternatively, it may include an agent that reduces the expression level of the gene encoding the protein.

The "pancreatic cancer" is divided into exocrine tumors and neuroendocrine tumors, and most types are exocrine tumor types, corresponding to pancreatic ductal adenocarcinoma (PDAC). These cancer cells or tissues of PDAC patients are cancer initiating cells with phenotypes of CD44(+)CD24(+) and CD44(+)CD24(+)EpCAM(+), which are chemically resistant, self-renewable and Because they have a subpopulation with differentiated capabilities, most of the cancer cells of the PDAC patients metastasize, and are highly resistant to classical treatments such as chemotherapy, radiation therapy, and immunotherapy. In particular, in the case of patients with cells having the CD44(+)CD24(+) phenotype, the prognosis is very poor compared to patients without it.

Pancreatic cancer of the present invention may be pancreatic ductal adenocarcinoma, but is not limited thereto.

The “metastasis” refers to cancer caused by cancer cells leaving a primary organ and moving to another organ and proliferating. The spread of cancer to other parts of the body can be largely divided into direct invasion of surrounding organs by the growth of cancer tissue from a primary cancer, and distant metastasis to other organs along blood vessels or lymphatic vessels.

The term “relapse” refers to a case where cancer reoccurs in the area where the cancer first occurred and then was determined to be cured due to treatment.

An agent that reduces the activity or expression of the protein may be a compound, a peptide mimetics, an aptamer, an antibody, or a natural product that specifically binds to the Jag1 protein or a part thereof.

An agent that reduces the expression level of the gene may be an antisense nucleotide, small interfering RNA (siRNA), short hairpin RNA (shRNA) or ribozyme that complementarily binds to the Jag1 gene or a part thereof.

Each of the above compositions may be formulated and used in the form of oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories or sterile injection solutions according to conventional methods.

The composition may be administered orally or parenterally, and in the case of parenteral administration, intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, etc. may be administered.

Solid preparations for oral administration include, but are not limited to, tablets, pills, powders, granules, capsules, and the like. Such a solid preparation may be prepared by mixing at least one or more excipients, for example, starch, calcium carbonate, sucrose, lactose, gelatin, etc., in addition to the active ingredient. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. It may be prepared by adding various excipients, for example, wetting agents, sweeteners, aromatics, and preservatives, in addition to liquids and liquid paraffin for oral use.

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations and tablets. Propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used as non-aqueous solvents and suspending agents. As a base for suppositories, Witepsol, Macrosol, Tween 61, cacao butter, laurin paper, glycerogelatin, and the like may be used.

The composition may further include pharmaceutically acceptable additives.

The additives may further include suitable carriers, excipients and diluents commonly used in the preparation of the composition.

The composition may include a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier is one commonly used in formulation, and includes lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate , microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, but are not limited thereto no.

Suitable pharmaceutically acceptable carriers and agents are described in detail in Remington's Pharmaceutical Sciences (19th ed, 1995).

The appropriate dosage of the composition varies depending on factors such as formulation method, administration method, patient's age, weight, sex, medical condition, food, administration time, administration route, excretion rate and response sensitivity, and the degree of bone loss of the patient. , It can be appropriately selected by those skilled in the art, specifically 0.001 mg/kg to 500 mg/kg, and can be divided and administered once a day to several times as needed.

The composition may be prepared in unit dosage form or multi-dose by formulating using a pharmaceutically acceptable carrier and/or excipient according to a method that can be easily performed by those skilled in the art. It can be prepared by incorporating into a container. At this time, the dosage form may be in the form of a solution, suspension or emulsion in an oil or aqueous medium, or in the form of an extract, powder, granule, tablet or capsule, and may additionally contain a dispersing agent or stabilizer.

Subjects for administration of the composition of the present invention include humans, monkeys, cows, horses, pigs, sheep, chickens, cats, dogs, mice, rabbits, etc., but are not particularly limited thereto.

Another embodiment of the present invention provides a method for screening for a therapeutic agent for recurrence or metastasis of pancreatic cancer.

The method includes: 1) treating pancreatic cancer cells expressing Jag1 protein or a gene encoding the Jag1 protein with a candidate substance; and

2) measuring the activity or expression level of the Jag1 protein or the expression level of a gene encoding the protein in cancer cells treated with the candidate substance.

The candidate substance may be obtained through a synthetic or natural compound library, a biological library, a spatially addressable parallel solid phase or solution phase library, or the like.

The "screening" is to select a substance having a specific target property from a candidate group composed of various substances by a specific manipulation or evaluation method.

An agent for measuring the expression level of the protein may be an antibody, an oligopeptide, a ligand, a peptide nucleic acid (PNA), or an aptamer that specifically binds to the protein.

The "PNA (Peptide Nucleic Acid)" refers to an artificially synthesized polymer similar to DNA or RNA, and was first introduced in 1991 by Professors Nielsen, Egholm, Berg and Buchardt of the University of Copenhagen, Denmark. While DNA has a phosphate-ribose backbone, PNA has a repeated N-(2-aminoethyl)-glycine backbone linked by peptide bonds, which greatly increases its binding force and stability to DNA or RNA, making it a molecule It is used in biology, diagnostic assays and antisense therapies. PNA is described by Nielsen PE, Egholm M, Berg RH, Buchardt O (December 1991). "Sequence-selective recognition of DNA by strand displacement with a thymine- substituted polyamide". Science 254 (5037): 1497-1500.

An agent for measuring the expression level of the gene may be a primer, probe, or antisense nucleotide that specifically binds to the gene.

The "primer" is a fragment that recognizes a target gene sequence, and includes a forward and reverse primer pair, preferably a primer pair that provides analysis results having specificity and sensitivity. High specificity can be imparted when the nucleic acid sequence of the primer is a sequence that is inconsistent with the non-target sequence present in the sample, so that the primer amplifies only the target gene sequence containing a complementary primer binding site and does not cause non-specific amplification. .

The "probe" refers to a substance capable of specifically binding to a target substance to be detected in a sample, and means a substance capable of specifically confirming the presence of a target substance in a sample through the binding. The type of probe is not limited as a material commonly used in the art, but preferably may be peptide nucleic acid (PNA), locked nucleic acid (LNA), peptide, polypeptide, protein, RNA, or DNA. More specifically, the probe is a biomaterial, including one derived from or similar to or produced in vitro, such as enzymes, proteins, antibodies, microorganisms, animal and plant cells and organs, nerve cells, DNA, and RNA, DNA includes cDNA, genomic DNA, and oligonucleotides, RNA includes genomic RNA, mRNA, and oligonucleotides, and examples of proteins may include antibodies, antigens, enzymes, peptides, and the like.

Another embodiment of the present invention provides a composition for diagnosing recurrence or metastasis of pancreatic cancer.

The composition may include an agent for measuring the expression level of the Jag1 protein or the gene encoding the same.

The above “diagnosis” means confirming the presence or characteristics of a pathological condition. For purposes of the present invention, diagnosis is to predict or confirm recurrence or metastasis following surgery or treatment of pancreatic cancer.

An agent for measuring the expression level of the protein may be an antibody, an oligopeptide, a ligand, a peptide nucleic acid (PNA), or an aptamer that specifically binds to the protein.

An agent for measuring the expression level of the gene may be a primer, probe, or antisense nucleotide that specifically binds to the gene.

Another embodiment of the present invention provides a kit for diagnosing recurrence or metastasis of cancer comprising the composition for diagnosis.

The "kit" refers to a tool capable of evaluating the expression level of a biomarker by labeling a probe or antibody that specifically binds to a biomarker component with a detectable label. Not only direct labeling of a detectable substance in relation to a probe or antibody by reaction with a substrate, but also indirect labeling in which a color-developing label is conjugated by reactivity with another directly labeled reagent. It may include a color-developing substrate solution, a washing solution, and other solutions that will undergo a color-reacting reaction with the marker, and may be prepared including reagent components to be used. In the present invention, the kit may be a kit containing essential elements necessary for performing RT-PCR, and in addition to each primer pair specific for the marker gene, a test tube, reaction buffer, deoxynucleotides (dNTPs), Taq-polymerization enzymes, reverse transcriptase, DNase, RNase inhibitors, sterile water, and the like. In addition, the kit may be a kit for detecting a gene for predicting HPD prognosis, including essential elements necessary for performing DNA chip. The DNA chip kit includes a substrate to which a cDNA corresponding to a gene or a fragment thereof is attached as a probe, and the substrate may include a cDNA corresponding to a quantitative control gene or a fragment thereof. The kit of the present invention is not limited thereto, as long as it is known in the art.

The kit may be an RT-PCR kit, a DNA chip kit, an ELISA kit, a protein chip kit, a rapid kit, or a multiple reaction monitoring (MRM) kit.

The kit may further include one or more other component compositions, solutions or devices suitable for the assay method.

Another embodiment of the present invention provides a method for providing information for diagnosing recurrence or metastasis of pancreatic cancer.

The method may include measuring the expression level of the Jag1 protein or a gene encoding the Jag1 protein in a biological sample isolated from a subject of interest.

According to the method, when the expression level of the Jag1 protein or the gene encoding the Jag1 protein measured in the biological sample is higher than that of the control group, it can be predicted that the possibility of recurrence or metastasis of cancer to the target subject is high.

The "object of interest" is an individual who has or has a high possibility of developing cancer, and may be mammals including humans, such as humans, rats, mice, guinea pigs, rabbits, monkeys, cats, dogs, and cows. , It may be selected from the group consisting of horses, pigs, sheep and goats, but is not limited thereto.

The "biological sample" refers to any material, biological fluid, tissue, or cell obtained from or derived from an individual, including whole blood, leukocytes, and peripheral blood mononuclear cells. , leukocyte buffy coat, plasma, serum, sputum, tears, mucus, nasal washes, nasal aspirate, respiration ( breath, urine, semen, saliva, peritoneal washings, ascites, cystic fluid, meningeal fluid, amniotic fluid, Glandular fluid, pancreatic fluid, lymph fluid, pleural fluid, nipple aspirate, bronchial aspirate, synovial fluid, joint aspirate ( joint aspirate), organ secretions, cells, cell extracts, and cerebrospinal fluid, but may be one or more selected from the group consisting of, but is not limited thereto.

Measurement of the expression level of the protein is protein chip analysis, immunoassay, ligand binding assay, MALDI-TOF (Matrix Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry) analysis, SELDI-TOF (Sulface Enhanced Laser Desorption/Ionization Time of Flight Mass Spectrometry) assay, radioimmunoassay, radioimmunodiffusion method, Oukteroni immunodiffusion method, rocket immunoelectrophoresis, tissue immunostaining, complement fixation assay, two-dimensional electrophoretic assay, liquid chromatography-mass spectrometry -Mass Spectrometry, LC-MS), LC-MS/MS (liquid chromatography-Mass Spectrometry/ Mass Spectrometry), Western blotting, or ELISA (enzyme linked immunosorbentassay).

In addition, the expression level of the protein may be measured by a multiple reaction monitoring (MRM) method.

In the multiple reaction monitoring method, a synthetic peptide or E. coli beta-galactosidase in which a specific amino acid constituting the target peptide is substituted with an isotope may be used as an internal standard material.

Measurement of the expression level of the gene encoding the protein is reverse transcription polymerase reaction (RT-PCR), competitive reverse transcription polymerase reaction (Competitive RT-PCR), real-time reverse transcription polymerase reaction (Real-time RT-PCR), RNase protection It may be performed by RNase protection assay (RPA), Northern blotting, or a DNA chip.

In one embodiment, an agent that reduces the activity or expression level of Jagged1 protein; Alternatively, it provides a composition for inhibiting plasticity of cancer initiating cells (CIC), comprising as an active ingredient an agent that reduces the expression level of a gene encoding the protein. In the above embodiment, the cancer is lung cancer, stomach cancer, ovarian cancer, cervical cancer, breast cancer, pancreatic cancer, colon cancer, colon cancer, esophageal cancer, skin cancer, thyroid cancer, kidney cancer, liver cancer, head and neck cancer, bladder cancer, prostate cancer, blood cancer, multiple Provides a composition that is at least one selected from the group consisting of myeloma, acute myelogenous leukemia, malignant lymphoma, thymic cancer, osteosarcoma, fibrotic tumor and brain cancer, wherein the cancer is pancreatic cancer, provides a composition, and the expression level of the gene The reducing agent is any one or more selected from the group consisting of antisense nucleotides, small interfering RNA (siRNA), short hairpin RNA (shRNA) and ribozymes that complementarily bind to the Jagged1 gene or a part thereof, and provides a composition, wherein The agent for reducing the activity or expression of a protein is at least one selected from the group consisting of compounds, peptide mimetics, aptamers, antibodies, and natural products that specifically bind to the Jagged1 protein or a portion thereof, and provides a composition. However, depending on the type of the agent, a selective effect may appear depending on each cancer type (see Mol Cancer Ther; 18(11) November 2019).

In one embodiment, an agent that reduces the activity or expression level of Jagged1 protein; Or it provides a composition for preventing or treating cancer recurrence or metastasis, comprising an agent that reduces the expression level of the gene encoding the protein as an active ingredient. In the above embodiment, the cancer is lung cancer, stomach cancer, ovarian cancer, cervical cancer, breast cancer, pancreatic cancer, colon cancer, colon cancer, esophageal cancer, skin cancer, thyroid cancer, kidney cancer, liver cancer, head and neck cancer, bladder cancer, prostate cancer, blood cancer, multiple Provides a composition that is at least one selected from the group consisting of myeloma, acute myelogenous leukemia, malignant lymphoma, thymic cancer, osteosarcoma, fibrotic tumor and brain cancer, wherein the cancer is pancreatic cancer, and the expression level of the gene The reducing agent is any one or more selected from the group consisting of antisense nucleotides, small interfering RNA (siRNA), short hairpin RNA (shRNA) and ribozymes that complementarily bind to the Jagged1 gene or a part thereof, and provides a composition, wherein The agent for reducing the activity or expression of a protein is at least one selected from the group consisting of compounds, peptide mimetics, aptamers, antibodies, and natural products that specifically bind to the Jagged1 protein or a portion thereof, and provides a composition.

In one embodiment, 1) treating cancer cells expressing Jagged1 protein or a gene encoding the same with a candidate substance; and 2) measuring the activity or expression level of Jagged1 protein or measuring the expression level of a gene encoding the protein in cancer cells treated with the candidate substance, suppressing plasticity of cancer or recurrence or metastasis of cancer A therapeutic screening method is provided. In the above embodiment, the cancer is lung cancer, stomach cancer, ovarian cancer, cervical cancer, breast cancer, pancreatic cancer, colon cancer, colon cancer, esophageal cancer, skin cancer, thyroid cancer, kidney cancer, liver cancer, head and neck cancer, bladder cancer, prostate cancer, blood cancer, multiple Provides a method of at least one selected from the group consisting of myeloma, acute myelogenous leukemia, malignant lymphoma, thymic cancer, osteosarcoma, fibrotic tumor and brain cancer, wherein the cancer is pancreatic cancer, and measurement after treatment with the candidate substance When the activity or expression level of the Jagged1 protein is decreased, or the expression level of the gene encoding the protein is decreased, the step of determining the candidate substance as a drug for cancer recurrence or metastasis treatment is provided. do.

In one embodiment, a composition for suppressing cancer plasticity or diagnosing cancer recurrence or metastasis, including an agent for measuring the expression level of Jagged1 protein or a gene encoding the same, is provided. In the above embodiment, the cancer is lung cancer, stomach cancer, ovarian cancer, cervical cancer, breast cancer, pancreatic cancer, colon cancer, colon cancer, esophageal cancer, skin cancer, thyroid cancer, kidney cancer, liver cancer, head and neck cancer, bladder cancer, prostate cancer, blood cancer, multiple Provides a composition that is at least one selected from the group consisting of myeloma, acute myeloid leukemia, malignant lymphoma, thymic cancer, osteosarcoma, fibrotic tumor and brain cancer, wherein the cancer is pancreatic cancer, and the expression level of the protein The agent to be measured provides a composition comprising at least one selected from the group consisting of an antibody, an oligopeptide, a ligand, a peptide nucleic acid (PNA), and an aptamer that binds specifically to the protein, and the gene An agent for measuring the expression level of provides a composition comprising at least one selected from the group consisting of primers, probes, and antisense nucleotides that specifically bind to the gene.

In one embodiment, a kit for suppressing plasticity of cancer or diagnosing cancer recurrence or metastasis, including the composition, is provided.

In one embodiment, a method for providing information for diagnosing cancer recurrence or metastasis, comprising measuring the expression level of Jagged1 protein or a gene encoding the same, in a biological sample isolated from a subject of interest is provided. In the above embodiment, the cancer is lung cancer, stomach cancer, ovarian cancer, cervical cancer, breast cancer, pancreatic cancer, colon cancer, colon cancer, esophageal cancer, skin cancer, thyroid cancer, kidney cancer, liver cancer, head and neck cancer, bladder cancer, prostate cancer, blood cancer, multiple Provides a method of at least one selected from the group consisting of myeloma, acute myelogenous leukemia, malignant lymphoma, thymic cancer, osteosarcoma, fibrotic tumor, and brain cancer, wherein the cancer is pancreatic cancer, and the method measured in the biological sample When the expression level of the Jagged1 protein or the gene encoding it is higher than that of the control group, it is predicted that the recurrence or metastasis of cancer to the subject is highly likely.

In one embodiment, a device for suppressing plasticity of cancer or diagnosing recurrence or metastasis of cancer, comprising: (a) a sample unit including a biological sample; (b) a measurement unit for receiving a sample from the sample unit and measuring the expression level of Jagged1 protein or a gene encoding the same; And (c) if the measurement result from the measurement unit of (b) is higher than that of the control group, a calculation unit that determines whether or not plasticity of cancer is suppressed or whether cancer recurrence or metastasis is highly likely to occur in the target subject. diagnostic equipment is provided.

The composition for inhibiting cancer-initiating cell reprogramming containing the Jagged1 protein inhibitor of the present invention inhibits the expression and activity of Jag1, an inducer of pancreatic cancer cell plasticity, thereby inhibiting cancer-initiating cells involved in tumor growth, induction of drug resistance, or recurrence of cancer. Since it can reduce plasticity, it can be used as a cancer-initiating cell-targeted therapy for pancreatic cancer.

Figure 1 shows the morphology of control and differentiated pancreatic cancer organoids.
Figure 2 shows the results of confirming the cell phenotypes of control and differentiated pancreatic cancer organoids through microscopy or flow cytometry.
3 is a graph showing the cell populations of CD24(+)CD44(-)EpCAM(+) and CD24(+)CD44(+)EpCAM(+) in control and differentiated pancreatic cancer organoids.
4 is a graph showing CD44, CA2 and CEL mRNA levels in control and differentiated pancreatic cancer organoids.
5 is a picture confirming the expression of CA2 in control and differentiated pancreatic cancer organoids.
6 is an image of a tumor mass, and the right side is a photograph of H&E staining.
7 is a FACS plot showing the CD24(+)CD44(+) and CD24(+)CD44(+)EpCAM(+) cancer initiating cell populations in the tumor mass.
Figure 8 is cryosection images of m/hCD31 (green), hCD44 (red) and hCD24 (white).
Figure 9 is a graph analyzing the cancer-initiating cell population after 7-day culture of an assembly including (I) differentiated pancreatic cancer cell (diPDAC) and (II) diPDAC/EC/autoimmune cell population.
10 shows the results of analyzing the level of JAG1 mRNA in normal pancreas of the GTEX data set and PAAD of the TCGA data set using recount3.
Figure 11 left shows CD24(+)CD44(-)EpCAM(+)JAG1(+) and CD24(+)CD44(+)EpCAM(+)JAG1(+) population analysis of human pancreatic cancer organoids and differentiated organoids. This is the FACS plot result shown.
FIG. 11 right shows CD24(+)CD44(-)EpCAM(+)JAG1(+) and CD24(+)CD44(+)EpCAM(+)JAG1(+) population analysis of human pancreatic cancer organoids and differentiated organoids. It is a quantified graph.
12 Left shows CFSE(-)CD24(+)CD44(+) cancer initiating cells in co-culture of differentiated CD44(-) cancer cells of CFSE-labeled HUVECs and pancreatic cancer organoids after treatment with control IgG and JAG1 neutralizing antibody. This is the FACS plot result.
Fig. 12 Right shows the quantification results of CFSE(-)CD24(+)CD44(+)CIC in the indicated groups.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for explaining the present invention in more detail, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention. .

Experiment method

[Test method 1] Pancreatic ductal adenocarcinoma patient sample

Pancreatic ductal adenocarcinoma (PDAC) tissue and endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) samples were obtained from Severance Hospital, Yonsei University College of Medicine. All human experiments related to this application were approved by the institutional review board (IRB) of Severance Hospital, and informed consent was obtained from the donor pancreatic cancer patient in accordance with the informed consent guidelines of the IRB.

[Experiment method 2] Production of human-derived pancreatic cancer organoids

The tissue sample obtained in [Test Method 1] was cultured in DMEM (Dulbecco's Modified Eagle's medium) medium containing 2 mg/ml of collagenase D (Sigma Aldrich, catalog number: 11088866001) and 10% FBS (Fetal bovine serum). , and sufficiently pulverized and decomposed through the process of incubating at 37 ° C. for 90 minutes. Then, the disaggregated tissue was washed with DPBS (Distilled phosphate buffer saline) containing 10% FBS (Fetal Bovine Serum), and growth factor reduce (GFR) Matrigel (Corning, catalog number: 356231) embedded in. Here, in the case of the EUS-FNA sample, a small amount of cellular material was collected by centrifugation at 400 × g for 10 minutes, and the EUS-FNA sample was lysed using RBC lysis buffer (QIAGen, catalog number: 158904) and washed twice using adDMEM/F12 medium (Invitrogen, catalog number: 12634028).

Thereafter, the matrix is polymerized through incubation in an incubator for 10 minutes, and pancreatic cancer organoid culture medium (medium 1) additionally containing the components listed in Table 1 below is added to adDMEM/F12. added and cultured.

ingredient manufacturing company Glutamax Invitrogen Antibiotic
(penicillin/streptomycin) Thermo Fisher Scientific B27 Thermo Fisher Scientific 1 mM, N-acetyl-L-cysteine
(N-acetyl-L-cysteine) Sigma Aldrich 50% v/v, Wnt3a-conditioned medium
(wnt3a-conditioned medium) 10% v/v, RSPO1-conditioned medium R&D system 100 ng/ml, Noggin recombinant protein Pepprotech
(Peptrotech) 50 ng/ml, epidermal growth factor
(Epidermal growth factor) Pepprotech 10 nM, Gastrin Sigma Aldrich 100 ng/ml, fibroblast growth factor 10 Pepprotech 10 mM, nicotinamide Sigma Aldrich 0.5 µM, A83-01 Tocris 10 µM, SB202190 Sigma Aldrich 1 μM, PGE2 Pepprotech

For tubular epithelial carcinoma cell differentiation, organoids were washed with basal medium and then cultured in AdDMEM/supplemented with 50 ng/ml EGF, Wnt-C59 (100 nM, Tocris) and DAPT (20 μM, Sigma-Aldrich), B27 for 2 days. Cultured in F12 medium.

[Experimental Method 3] PBMC Freezing and Separation

Peripheral blood mononuclear cells (PBMCs) were isolated from pancreatic cancer organoid pair blood using Ficoll-Paque. Isolated PBMCs were cryopreserved in FBS with 10% DMSO until analysis.

[Experiment method 4] HUVECs culture method

Human umbilical vein endothelial cells (HUVECs) were aliquoted into culture dishes coated with 1% gelatin, and EBM-2-based medium containing 1×EGM-2MV singlequots supplement pack (catalog number: CC-4147). (Lonza, catalog number: CC-3156).

[Experimental Method 5] Pancreatic cancer organoids, HUVECs and PBMCs co-culture method

After dissociating the pancreatic cancer organoids prepared in [Test Method 2] using TypLE Express, organoid-derived CD44(-) cancer cells were stained using CD44-APCcy7, and then flow cytometric sorting (FACS Aria III) was used. and classified. For cell tracking in flow cytometry, HUVECs and PBMCs were stained using CFSE according to the manufacturer's protocol. Then, organoid-derived differentiated CD44(-) cancer cells, HUVECs, and organoid-paired PBMCs were mixed at a ratio of 1:2:2. Thereafter, the cell mixture was placed in round-bottom ultra-low attachment plates (corning), medium 1 containing 10% Matrigel was added, and centrifuged at 100 × g for 3 minutes, , and further cultured for 1 day. The cell mixture was then cultured in basal medium (adDMEM/F12 medium containing GlutaMax and 5% FBS).

[Experiment method 6] Flow cytometry

Cells and tumor masses separated from spheroids by enzymes were stained using CD44-APCcy7 (Biolegend), CD24-Pecy7, EpCAM-PerCPcy5.5 and Jagged1-PE, followed by flow cytometry using LSR Ⅱ , analyzed through FlowJo and DIVA software.

[Experimental Method 7] Xenotransplantation

Animal experiments were performed with the approval of the Animal Care Committee of the Institute of Animal Research, Yonsei University College of Medicine. Cells isolated from the two PDAC assemblies were transplanted into NSG-(KbDb)null(IA)null (Jackson Laboratory) mice via subcutaneous injection using 50% Matrigel. After 6 weeks, the tumor was resected and frozen sections and paraffin sections were prepared. Then, immunofluorescence and H&E staining were performed using each frozen section and paraffin section.

[Experiment method 8] Immunofluorescence staining

The tumor tissue sections obtained in Experimental Method 7 were fixed using 2% paraformaldehyde, permeated with sucrose, and embedded through an OCT compound. For immunostaining, sections were blocked with BSA and stained with anti-CD31, anti-CD44 and anti-CD24. Samples were mounted via fluorescence mounting solution (DAKO). To evaluate the differentiation ability of organoids, differentiated organoids and control organoids were stained as follows. Differentiated organoids and control organoids were fixed with paraformaldehyde and glutaraldehyde for 10 minutes, washed with PBS supplemented with 10 mM NaBH4, and organoids were stained with anti-CD44 and anti-CD24. The organoids were stained with a secondary antibody and DAPI (4,6-diamidino-2-phenylindole), and images were obtained using a Zeiss LSM710 laser scan confocal local microscope.

[Experimental Method 9] Neutralizing JAG1

Organoid-derived CD44(-)PI(-) differentiated tubular epithelial cancer cells were sorted using flow cytometry sorting (FACS Aria III). Then, spheroid CD44(-)PI(-) differentiated tubular epithelial cancer cells were established with CFSE-labeled endothelial cells on a round-bottom ultra-low pressure attachment plate (Corning), and 10 ug/ml Jagged1 neutralizing antibody (R&D Systems , MAB12771) and control mouse IgG2b (R&D Systems, MAB004). After 1 day, the aggregates were embedded in GFR Matrigel, washed, and cultured in a basal medium containing Jagged1 neutralizing antibody or mouse IgG2b. After 6 days, the cancer-initiating cell population of the spheroids was analyzed using FACS analysis.

[Experiment method 10] Statistical analysis

Experimental results were calculated to be presented as mean ± SEM values. The Mann-Whitney U test was used for statistical comparison of two different samples, and Dunnett's multiple comparison test was used to evaluate statistical significance by comparing multiple samples. A statistically significant value was set to less than 0.05, and a statistical test was performed using the statistical package SPSS version 25 (SPSS Inc., Chicago, IL) and Graphpad Prism 8.

Experiment result

[Experiment Result 1] Confirmation of differentiation type of pancreatic cancer organoid cancer-initiating cells

As a result of confirming the differentiation morphology of the pancreatic cancer organoids in [Test Method 2], as shown in FIG. 1, the morphology of the organoids was changed to a cystic (transparent lumen) dense structure.

As shown in FIGS. 2 and 3, cancer initiating cells having the CD24(+)CD44(+)EpCAM(+) phenotype, which accounted for 56% in the control organoid, decreased to 14% in the differentiated organoid, and CD24 ( Cancer cells with +)CD44(-)EpCAM(+) phenotype increased from 30% to 78%.

As shown in FIGS. 4 and 5 , the level of CA2, a ductal epithelial cell marker, was significantly increased in differentiated cancer cells, but there was no change in CEL, an acinar biomarker.

From the above results, it can be seen that cancer initiating cells of pancreatic cancer organoids differentiate into pancreatic ductal adenocarcinoma differentiated into tubular epithelial cancer cells through inhibition of EGF, FGF10 and WNT/Notch.

[Experiment Result 2] Confirmation of plasticity of differentiated tubular epithelial cancer cells

As a result of observing the phenotype of the cells of the tumor mass (FIG. 6) generated by xenotransplanting the diPDAC assembly into a mouse, cancer initiating cells having a CD24(+)CD44(+)EPCAM(+) phenotype could be identified in the tumor mass. (FIG. 7), and it was confirmed through immunofluorescence staining that CD24(+)CD44(+) cancer initiating cells interact with endothelial cells (FIG. 8).

In addition, as shown in FIG. 9, when the diPDAC assembly was cultured alone, the cancer initiating cell population was 1.9%, whereas in the diPDAC assembly co-cultured with endothelial cells and autoimmune cells, the cancer initiating cell population increased to 10.4%. did

From the above results, it can be seen that the diPDAC assembly, co-cultured with endothelial cells and autoimmune cells, induces plasticity to reprogram differentiated tubular epithelial cancer cells into cancer initiating cells.

[Experiment Result 3] Confirmation of the role of JAG1 in the plasticity of cancer cells

As shown in FIG. 10 , the expression of JAG1 was significantly increased in pancreatic cancer cells compared to normal pancreas. In addition, compared to 3% of the AG1(+)CD24(+)CD44(-)EpCAM(+) non-cancer initiating cell group of the control group (before differentiation), the JAG1(+)CD24(+) of the differentiated pancreatic cancer organoids It was confirmed that the CD44(-)EpCAM(+) non-cancer initiating cell group increased to 18% (FIG. 11).

As a result of confirming the effect of JAG1 on cancer plasticity, compared to the control group (18%), the CD24(+)CD44(+)EpCAM(+) cancer initiating cell group in the JAG1 neutralization group decreased to 8% (FIG. 12).

Through the above results, it can be seen that JAG1 acts as a dynamic switch for cancer cell plasticity in pancreatic cancer organoids.

Having described specific parts of the present invention in detail above, it is clear that these specific techniques are only preferred embodiments for those skilled in the art, and the scope of the present invention is not limited thereto. Accordingly, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.

Claims (25)

agents that reduce the activity or expression level of Jagged1 protein; Or, a composition for inhibiting plasticity of cancer initiating cells (CIC) comprising, as an active ingredient, an agent that reduces the expression level of a gene encoding the protein.
According to claim 1,
The cancers include lung cancer, stomach cancer, ovarian cancer, cervical cancer, breast cancer, pancreatic cancer, colon cancer, colon cancer, esophageal cancer, skin cancer, thyroid cancer, kidney cancer, liver cancer, head and neck cancer, bladder cancer, prostate cancer, blood cancer, multiple myeloma, acute myeloid leukemia , At least one composition selected from the group consisting of malignant lymphoma, thymic cancer, osteosarcoma, fibrotic tumor and brain cancer.
According to claim 1,
Wherein the cancer is pancreatic cancer.
According to claim 1,
The agent that reduces the expression level of the gene is any one or more selected from the group consisting of antisense nucleotides, small interfering RNA (siRNA), short hairpin RNA (shRNA) and ribozymes that complementarily bind to the Jagged1 gene or a part thereof, composition.
According to claim 1,
The agent for reducing the activity or expression of the protein is at least one selected from the group consisting of compounds, peptide mimetics, aptamers, antibodies, and natural products that specifically bind to the Jagged1 protein or a part thereof, composition.
agents that reduce the activity or expression level of Jagged1 protein; Or, containing as an active ingredient an agent that reduces the expression level of the gene encoding the protein, A composition for preventing or treating cancer recurrence or metastasis.
According to claim 6,
The cancers include lung cancer, stomach cancer, ovarian cancer, cervical cancer, breast cancer, pancreatic cancer, colon cancer, colon cancer, esophageal cancer, skin cancer, thyroid cancer, kidney cancer, liver cancer, head and neck cancer, bladder cancer, prostate cancer, blood cancer, multiple myeloma, acute myeloid leukemia , At least one composition selected from the group consisting of malignant lymphoma, thymic cancer, osteosarcoma, fibrotic tumor and brain cancer.
According to claim 6,
Wherein the cancer is pancreatic cancer.
According to claim 6,
The agent that reduces the expression level of the gene is any one or more selected from the group consisting of antisense nucleotides, small interfering RNA (siRNA), short hairpin RNA (shRNA) and ribozymes that complementarily bind to the Jagged1 gene or a part thereof, composition.
According to claim 6,
The agent for reducing the activity or expression of the protein is at least one selected from the group consisting of compounds, peptide mimetics, aptamers, antibodies, and natural products that specifically bind to the Jagged1 protein or a part thereof, composition.
1) treating cancer cells expressing a Jagged1 protein or a gene encoding the same with a candidate substance; and
2) A method for screening cancer plasticity inhibitors or recurrence or metastasis drugs, comprising measuring the activity or expression level of Jagged1 protein or the expression level of a gene encoding the protein in cancer cells treated with the candidate substance .
According to claim 11,
The cancers include lung cancer, stomach cancer, ovarian cancer, cervical cancer, breast cancer, pancreatic cancer, colon cancer, colon cancer, esophageal cancer, skin cancer, thyroid cancer, kidney cancer, liver cancer, head and neck cancer, bladder cancer, prostate cancer, blood cancer, multiple myeloma, acute myeloid leukemia , At least one selected from the group consisting of malignant lymphoma, thymic cancer, osteosarcoma, fibrotic tumor and brain cancer, the method.
According to claim 11,
wherein the cancer is pancreatic cancer.
According to claim 11,
Further comprising determining the candidate substance as a drug for treating cancer recurrence and metastasis when the measured activity or expression level of the Jagged1 protein or the expression level of the gene encoding the protein is decreased after treatment with the candidate substance. , screening method.
A composition for diagnosing recurrence or metastasis of cancer, comprising an agent for measuring the expression level of a Jagged1 protein or a gene encoding the same.
According to claim 15,
The cancers include lung cancer, stomach cancer, ovarian cancer, cervical cancer, breast cancer, pancreatic cancer, colon cancer, colon cancer, esophageal cancer, skin cancer, thyroid cancer, kidney cancer, liver cancer, head and neck cancer, bladder cancer, prostate cancer, blood cancer, multiple myeloma, acute myeloid leukemia , At least one composition selected from the group consisting of malignant lymphoma, thymic cancer, osteosarcoma, fibrotic tumor and brain cancer.
According to claim 15,
Wherein the cancer is pancreatic cancer.
According to claim 15,
The agent for measuring the expression level of the protein comprises at least one selected from the group consisting of antibodies, oligopeptides, ligands, PNAs (peptide nucleic acids) and aptamers that bind specifically to the protein, composition .
According to claim 15,
An agent for measuring the expression level of the gene comprises at least one selected from the group consisting of primers, probes and antisense nucleotides that specifically bind to the gene, composition.
A kit for suppressing cancer plasticity or diagnosing cancer recurrence or metastasis, comprising the composition of any one of claims 15 to 19.
A method for providing information for suppressing plasticity of cancer or diagnosing recurrence or metastasis of cancer, comprising measuring the expression level of Jagged1 protein or a gene encoding the same in a biological sample isolated from a subject of interest.
According to claim 21,
The cancers include lung cancer, stomach cancer, ovarian cancer, cervical cancer, breast cancer, pancreatic cancer, colon cancer, colon cancer, esophageal cancer, skin cancer, thyroid cancer, kidney cancer, liver cancer, head and neck cancer, bladder cancer, prostate cancer, blood cancer, multiple myeloma, acute myeloid leukemia , At least one selected from the group consisting of malignant lymphoma, thymic cancer, osteosarcoma, fibrotic tumor and brain cancer, the method.
According to claim 21,
wherein the cancer is pancreatic cancer.
According to claim 21,
If the expression level of the Jagged1 protein or the gene encoding it measured in the biological sample is higher than that of the control group, it is predicted that the cancer recurrence or metastasis is highly likely to occur in the target subject.
As a device for suppressing plasticity of cancer or diagnosing recurrence or metastasis of cancer,
(a) a sample unit containing a biological sample;
(b) a measurement unit for receiving a sample from the sample unit and measuring the expression level of Jagged1 protein or a gene encoding the same; and
(c) if the measurement result from the measurement unit of (b) is higher than that of the control group, a calculation unit that determines whether or not plasticity of cancer is suppressed or whether cancer recurrence or metastasis is highly likely to occur in the target subject; diagnosis comprising device.

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