WO2023287079A1 - Biomarker composition for predicting triple negative breast cancer metastasis, containing csde1 as active ingredient - Google Patents

Biomarker composition for predicting triple negative breast cancer metastasis, containing csde1 as active ingredient Download PDF

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WO2023287079A1
WO2023287079A1 PCT/KR2022/009489 KR2022009489W WO2023287079A1 WO 2023287079 A1 WO2023287079 A1 WO 2023287079A1 KR 2022009489 W KR2022009489 W KR 2022009489W WO 2023287079 A1 WO2023287079 A1 WO 2023287079A1
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csde1
breast cancer
triple
negative breast
expression
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PCT/KR2022/009489
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French (fr)
Korean (ko)
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박종훈
김예솔
이수빈
고제영
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숙명여자대학교산학협력단
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Publication of WO2023287079A1 publication Critical patent/WO2023287079A1/en

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    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A61K31/7105—Natural ribonucleic acids, i.e. containing only riboses attached to adenine, guanine, cytosine or uracil and having 3'-5' phosphodiester links
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • 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
    • G01N33/57407—Specifically defined cancers
    • G01N33/57415—Specifically defined cancers of breast
    • 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
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/118—Prognosis of disease development
    • 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
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/136—Screening for pharmacological compounds
    • 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
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/158—Expression markers
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2500/00—Screening for compounds of potential therapeutic value
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis

Definitions

  • the present invention relates to a biomarker composition for predicting metastasis of triple negative breast cancer comprising CSDE1 as an active ingredient.
  • TNBC triple negative breast cancer
  • ER estrogen receptor
  • PR progesterone receptor
  • HER2 human epidermal growth factor receptor
  • TNBC patients are aggressive and have the worst prognosis.
  • Luminal A, Luminal B and HER2 Unlike other breast cancer subtypes, TNBC treatment is particularly difficult because of the lack of three hormone receptors that are therapeutic targets.
  • Prognostic indicators include common factors such as molecular markers, tumor size, lymph node status and histological grade, as well as providing some information about prognosis. Although these molecular markers and prognostic criteria provide some guidelines for predicting the fate of patients and selecting appropriate treatment methods, they are insufficient as methods to specifically and sensitively evaluate the prognosis of metastasis in triple-negative breast cancer, requiring the development of new methods. do.
  • An object of the present invention is to provide a biomarker composition for diagnosing triple negative breast cancer (TNBC) or predicting metastasis prognosis, comprising CSDE1 or a gene encoding the same as an active ingredient.
  • TNBC triple negative breast cancer
  • another object of the present invention is to provide a composition for diagnosis of triple-negative breast cancer or prediction of metastasis prognosis comprising an agent capable of measuring the expression level of CSDE1 as an active ingredient, and a kit for diagnosis of triple-negative breast cancer or prediction of metastasis prognosis containing the same.
  • Another object of the present invention is to provide a method for providing information for diagnosing triple negative breast cancer or predicting metastasis prognosis, comprising measuring the expression level of CSDE1 from a sample isolated from a patient.
  • Another object of the present invention is to provide a pharmaceutical composition for preventing, treating, or inhibiting metastasis of triple-negative breast cancer comprising a CSDE1 expression or activity inhibitor as an active ingredient.
  • Another object of the present invention is to provide a method for screening triple negative breast cancer therapeutics or metastasis inhibitors by measuring the expression level of CSDE1.
  • Another object of the present invention is to provide a reagent composition for inhibiting RAC1 expression in triple-negative breast cancer cells in vitro, comprising a CSDE1 expression or activity inhibitor as an active ingredient.
  • Another object of the present invention is to provide a method for inhibiting RAC1 expression comprising treating triple negative breast cancer cells with an inhibitor of CSDE1 expression or activity in vitro.
  • the present invention provides a biomarker composition for diagnosing triple negative breast cancer (TNBC) or predicting metastasis prognosis, comprising CSDE1 or a gene encoding the same as an active ingredient.
  • the present invention provides a composition for diagnosing triple-negative breast cancer or predicting the prognosis of metastasis, comprising an agent capable of measuring the expression level of CSDE1 as an active ingredient.
  • the present invention provides a kit for diagnosing triple-negative breast cancer or predicting the prognosis of metastasis comprising the composition.
  • the present invention (1) measuring the mRNA expression level of the CSDE1 gene or the expression level of the CSDE1 protein from a sample isolated from the patient; (2) comparing the mRNA expression level of the CSDE1 gene or the expression level of the CSDE1 protein with a control sample; and (3) determining triple-negative breast cancer when the mRNA expression level of the CSDE1 gene or the CSDE1 protein expression level is higher than that of the control sample.
  • the present invention comprises the steps of (1) measuring the mRNA expression level of the CSDE1 gene or the expression level of the CSDE1 protein from a sample isolated from a triple-negative breast cancer patient; (2) comparing the mRNA expression level of the CSDE1 gene or the expression level of the CSDE1 protein with a control sample; and (3) determining that the risk of triple-negative breast cancer metastasis is high when the mRNA expression level of the CSDE1 gene or the CSDE1 protein expression level is higher than that of a control sample.
  • the present invention provides a pharmaceutical composition for preventing, treating, or inhibiting metastasis of triple-negative breast cancer comprising a CSDE1 expression or activity inhibitor as an active ingredient.
  • the present invention comprises the steps of (1) contacting a test substance to triple-negative breast cancer cells; (2) measuring the expression or activity level of CSDE1 in triple-negative breast cancer cells contacted with the test substance; and (3) selecting a test substance having a reduced expression or activity level of the CSDE1 compared to a control sample.
  • the present invention comprises the steps of (1) contacting a test substance to triple-negative breast cancer cells; (2) measuring the expression or activity level of CSDE1 in triple-negative breast cancer cells contacted with the test substance; and (3) selecting a test substance having a reduced expression or activity of the CSDE1 compared to a control sample.
  • the present invention comprises the steps of (1) contacting a test substance to triple-negative breast cancer cells; (2) measuring the expression or activity levels of CSDE1 and RAC1 in triple-negative breast cancer cells contacted with the test substance; and (3) providing a screening method for triple-negative breast cancer metastasis inhibitors comprising the step of selecting a test substance in which the expression or activity of CSDE1 and RAC1 is reduced compared to a control sample.
  • the present invention provides a reagent composition for inhibiting RAC1 expression in triple-negative breast cancer cells in vitro, comprising a CSDE1 expression or activity inhibitor as an active ingredient.
  • the present invention provides a method for inhibiting RAC1 expression comprising treating triple-negative breast cancer cells with an inhibitor of CSDE1 expression or activity in vitro.
  • the present invention relates to a biomarker composition for predicting metastasis of triple-negative breast cancer comprising CSDE1 as an active ingredient. It was confirmed that That is, the present invention suggests a transcriptional regulatory mechanism of the Rac1 gene associated with cancer metastasis based on the expression of CSDE1 in triple-negative breast cancer. Using this, the present invention can be usefully utilized as a biomarker for predicting progression and metastasis of triple-negative breast cancer. In addition, using the biomarker of the present invention, it is possible to reduce unnecessary treatment and establish a treatment strategy tailored to the patient, and to reduce the treatment period and treatment cost through appropriate treatment suitable for triple-negative breast cancer patients without anticancer drugs capable of targeted treatment. Expect to be able to do it.
  • Figure 1 shows the results of confirming the increase in CSDE1 expression in a triple negative breast cancer (TNBC) model and verifying the possibility of using it as a metastasis/prognosis predictive biomarker.
  • TNBC triple negative breast cancer
  • Figure 2 shows the results of confirming the anticancer effect of triple-negative breast cancer cell lines through CSDE1 expression inhibition (siRNA, knock-out).
  • FIG. 3 shows the results of discovering RAC1, a cancer metastasis gene based on CSDE1 expression.
  • Figure 4 shows the results suggesting the mechanism of RAC1 transcriptional regulation by CSDE1.
  • Figure 5 shows the results of confirming the decrease in triple-negative breast cancer survival rate according to the increased expression of CSDE1 and RAC1.
  • the present invention provides a biomarker composition for diagnosis of triple negative breast cancer (TNBC) comprising CSDE1 or a gene encoding the same as an active ingredient.
  • TNBC triple negative breast cancer
  • the present invention provides a biomarker composition for predicting the prognosis of triple-negative breast cancer metastasis comprising CSDE1 or a gene encoding the same as an active ingredient.
  • the biomarker composition may further include RAC1 or a gene encoding the same, but is not limited thereto.
  • CSDE1 cold shock domain containing E1
  • NCBI accession no it may be NM_001007553.3, NM_007158.6, NM_001130523.3, NM_001242891.2, NM_001242892.2 or NM_001242893.2, but is not limited thereto.
  • RAC1 Ras-related C3 botulinum toxin substrate 1
  • NCBI accession no It may be NM_006908.5 or NM_018890.4, but is not limited thereto.
  • diagnosis refers to determining a subject's susceptibility to a specific disease or disorder, determining whether a subject currently has a specific disease or disorder, as long as he or she has a specific disease or disorder. determining a subject's prognosis, or therametrics (eg, monitoring a subject's condition to provide information about treatment efficacy).
  • prognosis in this specification refers to the outlook for future symptoms or progress determined by diagnosing a disease. In cancer patients, prognosis usually means metastasis or survival period within a certain period after cancer onset or surgical procedure. Prediction of prognosis is a very important clinical task because it provides clues to the direction of future triple-negative breast cancer treatment, including whether triple-negative breast cancer patients will undergo chemotherapy.
  • Metastasis refers to a state in which a certain tumor is transplanted from its primary site to another body part along various routes and settles and proliferates there. Metastasis of cancer is not only determined by the unique characteristics of the cancer, but also an event that is the most important clue in determining the prognosis of cancer, so it is treated as the most important clinical information related to the survival of cancer patients.
  • the present invention provides a composition for diagnosis of triple-negative breast cancer comprising an agent capable of measuring the expression level of CSDE1 as an active ingredient.
  • the present invention provides a composition for predicting the prognosis of triple-negative breast cancer metastasis, comprising an agent capable of measuring the expression level of CSDE1 as an active ingredient.
  • the biomarker composition may further include RAC1 or a gene encoding the same, but is not limited thereto.
  • the agent capable of measuring the expression level of the CSDE1 or RAC1 is a primer or probe that specifically binds to the CSDE1 or RAC1 gene, an antibody, peptide, or aptamer that specifically binds to the CSDE1 or RAC1 protein Or it may be a compound, but is not limited thereto.
  • the present invention provides a kit for diagnosing triple-negative breast cancer comprising the composition.
  • the present invention provides a kit for predicting the prognosis of triple-negative breast cancer metastasis comprising the composition.
  • primer refers to a nucleic acid sequence having a short free 3' hydroxyl group, capable of base pairing with a complementary template, and serving as a starting point for template strand copying. refers to the sequence of nucleic acids. Primers can initiate DNA synthesis in the presence of a reagent for polymerization (ie, DNA polymerase or reverse transcriptase) and four different nucleoside triphosphates in an appropriate buffer and temperature. PCR conditions and lengths of sense and antisense primers can be appropriately selected according to techniques known in the art.
  • probe refers to a nucleic acid fragment such as RNA or DNA corresponding to a few bases to several hundred bases in length that can specifically bind to mRNA and is labeled, so that the presence or absence of a specific mRNA, expression quantity can be checked.
  • the probe may be manufactured in the form of an oligonucleotide probe, a single strand DNA probe, a double strand DNA probe, an RNA probe, or the like. Selection of an appropriate probe and hybridization conditions can be appropriately selected according to techniques known in the art.
  • the term "antibody” is a term known in the art and refers to a specific immunoglobulin directed against an antigenic site.
  • the antibody in the present invention refers to an antibody that specifically binds to LRP-1 of the present invention, and the antibody can be prepared according to a conventional method in the art.
  • the type of antibody includes polyclonal antibodies or monoclonal antibodies, and all immunoglobulin antibodies are included.
  • the antibody is meant in its complete form with two full-length light chains and two full-length heavy chains.
  • the antibody also includes special antibodies such as humanized antibodies.
  • the kit of the present invention is an antibody that specifically binds to a marker component, a secondary antibody conjugate conjugated with a label that develops color by reaction with the substrate, a color-developing substrate solution that will react with the label, a washing solution, and It may contain an enzyme reaction stop solution, etc., and may be manufactured in a number of separate packaging or compartments containing reagent components to be used.
  • the term "peptide” has the advantage of having high binding ability to a target substance, and does not undergo denaturation even during heat/chemical treatment.
  • it can be attached to other proteins and used as a fusion protein. Specifically, since it can be used by attaching it to a polymer protein chain, it can be used as a diagnostic kit and a drug delivery material.
  • aptamer refers to a special kind of single-stranded nucleic acid (DNA, RNA or modified nucleic acid) having a stable tertiary structure and being able to bind to a target molecule with high affinity and specificity.
  • DNA DNA, RNA or modified nucleic acid
  • aptamers can specifically bind to antigenic substances in the same way as antibodies, but are more stable than proteins, have a simple structure, and are composed of polynucleotides that are easy to synthesize, so they can be used instead of antibodies.
  • the present invention (1) measuring the mRNA expression level of the CSDE1 gene or the expression level of the CSDE1 protein from a sample isolated from the patient; (2) comparing the mRNA expression level of the CSDE1 gene or the expression level of the CSDE1 protein with a control sample; and (3) determining triple-negative breast cancer when the mRNA expression level of the CSDE1 gene or the CSDE1 protein expression level is higher than that of the control sample.
  • the present invention comprises the steps of (1) measuring the mRNA expression level of the CSDE1 gene or the expression level of the CSDE1 protein from a sample isolated from a triple-negative breast cancer patient; (2) comparing the mRNA expression level of the CSDE1 gene or the expression level of the CSDE1 protein with a control sample; and (3) determining that the risk of triple-negative breast cancer metastasis is high when the mRNA expression level of the CSDE1 gene or the CSDE1 protein expression level is higher than that of a control sample.
  • the present invention (1) measuring the mRNA expression level of the CSDE1 gene and RAC1 gene or the expression level of the CSDE1 protein and RAC1 protein from a sample isolated from a triple-negative breast cancer patient; (2) comparing the mRNA expression level of the CSDE1 gene and the RAC1 gene or the expression level of the CSDE1 protein and the RAC1 protein with a control sample; and (3) determining that the risk of metastasis of triple-negative breast cancer is high when the mRNA expression levels of the CSDE1 gene and the RAC1 gene or the expression levels of the CSDE1 protein and the RAC1 protein are higher than those in the control sample.
  • the method for measuring the mRNA expression level is RT-PCR, competitive RT-PCR (Competitive RT-PCR), real-time RT-PCR (Real-time RT-PCR), RNase protection assay (RPA; RNase protection assay ), Northern blotting, and DNA chips, but are not limited thereto.
  • methods for measuring the protein expression level include Western blot, enzyme linked immunosorbent assay (ELISA), radioimmunoassay (RIA), radioimmunodiffusion, Ouchterlony immunodiffusion method, Rocket immunoelectrophoresis, tissue immunostaining, immunoprecipitation assay, complement fixation assay, FACS, and protein chip are used, but are not limited thereto.
  • ELISA enzyme linked immunosorbent assay
  • RIA radioimmunoassay
  • Ouchterlony immunodiffusion method Rocket immunoelectrophoresis, tissue immunostaining, immunoprecipitation assay, complement fixation assay, FACS, and protein chip are used, but are not limited thereto.
  • sample isolated from a patient means a sample such as tissue, cell, whole blood, serum, plasma, saliva, sputum, cerebrospinal fluid, or urine that is different from the control group in terms of the expression level of the CSDE1 or RAC1, which is a biomarker. Including, but not limited to.
  • the present invention provides a pharmaceutical composition for preventing or treating triple-negative breast cancer comprising a CSDE1 expression or activity inhibitor as an active ingredient.
  • the present invention provides a pharmaceutical composition for inhibiting metastasis of triple-negative breast cancer comprising a CSDE1 expression or activity inhibitor as an active ingredient.
  • the pharmaceutical composition may inhibit RAC1 expression, but is not limited thereto.
  • the CSDE1 protein expression inhibitor may be an antisense nucleotide, small interfering RNA (siRNA) or short hairpin RNA (shRNA) that binds complementarily to the mRNA of the CSDE1 gene
  • the CSDE1 Protein activity inhibitors may be compounds, peptides, peptide mimetics, aptamers, antibodies, or natural products that specifically bind to the CSDE1 protein, but are not limited thereto.
  • the pharmaceutical composition of the present invention may include chemical substances, nucleotides, antisense, siRNA oligonucleotides, and natural extracts as active ingredients.
  • the pharmaceutical composition or combined preparation of the present invention can be prepared using pharmaceutically suitable and physiologically acceptable adjuvants in addition to the active ingredients, and the adjuvants include excipients, disintegrants, sweeteners, binders, coating agents, expanding agents, and lubricants. , solubilizing agents such as lubricants or flavoring agents may be used.
  • the pharmaceutical composition of the present invention may be preferably formulated as a pharmaceutical composition by including one or more pharmaceutically acceptable carriers in addition to the active ingredient for administration.
  • acceptable pharmaceutical carriers are sterile and biocompatible, and include saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol and One or more of these components may be mixed and used, and other conventional additives such as antioxidants, buffers, and bacteriostatic agents may be added if necessary.
  • diluents, dispersants, surfactants, binders, and lubricants may be additionally added to prepare formulations for injections such as aqueous solutions, suspensions, and emulsions, pills, capsules, granules, or tablets.
  • the pharmaceutical formulation form of the pharmaceutical composition of the present invention may be granules, powders, coated tablets, tablets, capsules, suppositories, syrups, juices, suspensions, emulsions, drops or injectable solutions, and sustained-release preparations of active compounds.
  • the pharmaceutical composition of the present invention can be administered in a conventional manner via intravenous, intraarterial, intraperitoneal, intramuscular, intraarterial, intraperitoneal, intrasternal, transdermal, intranasal, inhalational, topical, rectal, oral, intraocular or intradermal routes.
  • An effective amount of the active ingredient of the pharmaceutical composition of the present invention means an amount required for preventing or treating a disease.
  • the type of disease the severity of the disease, the type and amount of the active ingredient and other ingredients contained in the composition, the type of formulation and the patient's age, weight, general health condition, sex and diet, administration time, administration route and composition It can be controlled by various factors including secretion rate, duration of treatment, and drugs used concurrently.
  • the composition of the present invention is administered once to several times a day, in the case of a compound, 0.1 ng / kg to 10 g / kg, polypeptide, It can be administered at a dose of 0.1 ng/kg to 10 g/kg for proteins or antibodies and 0.01 ng/kg to 10 g/kg for antisense nucleotides, siRNA, shRNAi, and miRNA.
  • the present invention comprises the steps of (1) contacting a test substance to triple-negative breast cancer cells; (2) measuring the expression or activity level of CSDE1 in triple-negative breast cancer cells contacted with the test substance; and (3) selecting a test substance having a reduced expression or activity level of the CSDE1 compared to a control sample.
  • the present invention comprises the steps of (1) contacting a test substance to triple-negative breast cancer cells; (2) measuring the expression or activity level of CSDE1 in triple-negative breast cancer cells contacted with the test substance; and (3) selecting a test substance having a reduced expression or activity of the CSDE1 compared to a control sample.
  • the present invention comprises the steps of (1) contacting a test substance to triple-negative breast cancer cells; (2) measuring the expression or activity levels of CSDE1 and RAC1 in triple-negative breast cancer cells contacted with the test substance; and (3) providing a screening method for triple-negative breast cancer metastasis inhibitors comprising the step of selecting a test substance in which the expression or activity of CSDE1 and RAC1 is reduced compared to a control sample.
  • test substance used while referring to the screening method of the present invention refers to an unknown candidate substance used in screening to test whether it affects the expression level of a gene or affects the expression or activity of a protein. do.
  • the samples include, but are not limited to, chemical substances, nucleotides, antisense-RNA, small interference RNA (siRNA), and natural product extracts.
  • the present invention provides a reagent composition for inhibiting RAC1 expression in triple-negative breast cancer cells in vitro, comprising a CSDE1 expression or activity inhibitor as an active ingredient.
  • the present invention provides a method for inhibiting RAC1 expression comprising treating triple-negative breast cancer cells with an inhibitor of CSDE1 expression or activity in vitro.
  • TNBC_TCGA data analysis Through cBioportal and TCGA breast cancer dataset analysis, breast cancer subtype-specific CSDE1 gene expression was compared and analyzed.
  • qRT-PCR analysis mRNA was extracted using Nucleospin kit to confirm the expression change of a specific gene. After synthesizing the extracted mRNA into cDNA using reverse transcriptase (RTase), qRT-PCR analysis was performed using a primer for a specific gene.
  • RTase reverse transcriptase
  • Cell viability assay Cells were seeded in 96-well with the same number of cells. After inoculation, cell viability was measured at 24-hour intervals using an ELISA device using CCK-8 (cell counting kit-8) reagent.
  • Transwell migration assay After inserting Transwell insert into 24-well, cells suspended in serum-free medium were inoculated into the insert, and 10% FBS medium was placed under the insert membrane, Migrated cells were observed by staining with crystal violet reagent.
  • Transwell invasion assay Matrigel-coated transwell insert was placed in 24-well, cells suspended in serum-free medium were inoculated into the insert, and 10% FBS medium was placed under the insert membrane. After putting in, the invaded cells were observed through crystal violet reagent staining.
  • IP Immunoprecipitation
  • RNA-IP immunoprecipitation
  • CSDE1 expression which is increased in triple-negative breast cancer, could affect the regulation of RAC1 expression, a cancer metastasis gene.
  • CSDE1 expression was reduced in triple-negative breast cancer cell lines (MDA-MB-231, Hs578T) through siRNA transfection, it was confirmed that both mRNA and protein expression of RAC1 were reduced (FIGS. 3A and 3B).
  • RAC1 protein expression was decreased in the TNBC cell line in which the CSDE1 gene was knocked out (KO) (FIG. 3C).
  • CSDE1 expression was increased in normal cell lines MCF10A and HEK293T cells through plasmid transfection, it was confirmed that RAC1 expression increased (FIG. 3D).
  • CSDE1 In the regulation of RAC1 by CSDE1, it was verified which step in the level of transcription regulation of RAC1 specifically acts. First, the interaction between CSDE1 and RNA polymerase II p-CTD (ser2), RNA polymerase II p-CTD (ser5), and RNA polymerase II p-CTD (ser7) was examined through immunoprecipitation (IP). As a result of confirmation, it was confirmed that CSDE1 binds to each RNA pol II (FIG. 4A). Considering that ser5 and ser7 of RNA pol II are used as initiation and elongation markers at the level of transcriptional regulation, and ser2 is used as a termination marker, CSDE1 has a role in transcriptional regulation of specific genes.

Abstract

The present invention relates to a biomarker composition for predicting triple negative breast cancer metastasis, containing CSDE1 as an active ingredient, and it has been identified that CSDE1 gene is highly expressed in triple negative breast cancer and regulates the transcription of Rac1, which is a gene associated with invasion and metastasis. The present invention provides a mechanism for regulating the transcription of cancer metastasis-associated Rac1 gene on the basis of the expression of CSDE1 in triple negative breast cancer. Therefore, the present invention can be effectively used as a biomarker for predicting the progression and metastasis of triple negative breast cancer. In addition, by using a biomarker of the present invention, it is expected that unnecessary treatment can be reduced, and patient-specific treatment strategies can be established, and treatment periods and treatment costs can be reduced through appropriate treatment for patients with triple negative breast cancer for which there are no anticancer agents capable of providing targeted therapy.

Description

CSDE1을 유효성분으로 포함하는 삼중음성유방암 전이 예측용 바이오마커 조성물Biomarker composition for predicting metastasis of triple-negative breast cancer containing CSDE1 as an active ingredient
본 발명은 CSDE1을 유효성분으로 포함하는 삼중음성유방암의 전이 예측용 바이오마커 조성물에 관한 것이다.The present invention relates to a biomarker composition for predicting metastasis of triple negative breast cancer comprising CSDE1 as an active ingredient.
전세계적으로 유방암은 여성에게 가장 흔한 암으로서, 이 중에서 삼중음성유방암(Triple Negative Breast Cancer; TNBC)은 전체 유방암 환자의 12-20%를 차지한다. TNBC는 유방암 서브타입인데, 3가지 호르몬성 멤브레인 수용체인 에스트로겐 수용체(estrogen receptor; ER), 프로게스테론 수용체(progesterone receptor, PR) 및 인간 상피 성장인자 수용체(human epidermal growth factor receptor; HER2)가 존재하지 않는다. 유방암 서브타입 중에서도 TNBC 환자들은 공격적이고 가장 나쁜 예후를 나타낸다. 다른 유방암 서브타입들(Luminal A, Luminal B 및 HER2)과 달리, TNBC 치료는 치료 표적인 3가지 호르몬 수용체가 없기 때문에 특히 어렵다.Worldwide, breast cancer is the most common cancer in women, among which triple negative breast cancer (TNBC) accounts for 12-20% of all breast cancer patients. TNBC, a subtype of breast cancer, lacks the three hormonal membrane receptors: estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor (HER2) . Among breast cancer subtypes, TNBC patients are aggressive and have the worst prognosis. Unlike other breast cancer subtypes (Luminal A, Luminal B and HER2), TNBC treatment is particularly difficult because of the lack of three hormone receptors that are therapeutic targets.
현재까지 삼중음성유방암의 전이 예측과 관련된 대부분의 연구는 삼중음성유방암 예후를 분석하고 치료반응을 예측하기 위한 방법 및 인자의 동정에 집중되어 있다. 예후 표시자(indicator)는 분자 마커와 같은 통상적인 인자를 포함하며, 종양 크기, 림프절 상태 및 조직학적 등급뿐만 아니라 예후에 대한 일부 정보를 제공한다. 상기와 같은 분자 마커 및 예후 기준은 환자의 운명을 예측하고 적절한 치료방법을 선정하는데 일부 지침을 제공하나, 삼중음성유방암의 전이 예후를 특이적이고 민감하게 평가하는 방법으로서는 부족하여 새로운 방법의 개발이 요구된다.Until now, most studies related to the prediction of metastasis of triple-negative breast cancer have focused on identifying methods and factors for analyzing the prognosis of triple-negative breast cancer and predicting treatment response. Prognostic indicators include common factors such as molecular markers, tumor size, lymph node status and histological grade, as well as providing some information about prognosis. Although these molecular markers and prognostic criteria provide some guidelines for predicting the fate of patients and selecting appropriate treatment methods, they are insufficient as methods to specifically and sensitively evaluate the prognosis of metastasis in triple-negative breast cancer, requiring the development of new methods. do.
본 발명의 목적은 CSDE1 또는 이를 코딩하는 유전자를 유효성분으로 포함하는 삼중음성유방암(Triple Negative Breast Cancer; TNBC) 진단 또는 전이 예후 예측용 바이오마커 조성물을 제공하는데 있다.An object of the present invention is to provide a biomarker composition for diagnosing triple negative breast cancer (TNBC) or predicting metastasis prognosis, comprising CSDE1 or a gene encoding the same as an active ingredient.
또한, 본 발명의 다른 목적은 CSDE1의 발현수준을 측정할 수 있는 제제를 유효성분으로 포함하는 삼중음성유방암 진단 또는 전이 예후 예측용 조성물, 이를 포함하는 삼중음성유방암 진단용 또는 전이 예후 예측용 키트를 제공하는데 있다.In addition, another object of the present invention is to provide a composition for diagnosis of triple-negative breast cancer or prediction of metastasis prognosis comprising an agent capable of measuring the expression level of CSDE1 as an active ingredient, and a kit for diagnosis of triple-negative breast cancer or prediction of metastasis prognosis containing the same. are doing
또한, 본 발명의 또 다른 목적은 환자에서 분리된 시료로부터 CSDE1의 발현수준을 측정하는 단계를 포함하는 삼중음성유방암 진단 또는 전이 예후 예측을 위한 정보를 제공하는 방법을 제공하는데 있다.Another object of the present invention is to provide a method for providing information for diagnosing triple negative breast cancer or predicting metastasis prognosis, comprising measuring the expression level of CSDE1 from a sample isolated from a patient.
또한, 본 발명의 또 다른 목적은 CSDE1 발현 또는 활성 억제제를 유효성분으로 포함하는 삼중음성유방암 예방, 치료 또는 전이 억제용 약학조성물을 제공하는데 있다.Another object of the present invention is to provide a pharmaceutical composition for preventing, treating, or inhibiting metastasis of triple-negative breast cancer comprising a CSDE1 expression or activity inhibitor as an active ingredient.
또한, 본 발명의 또 다른 목적은 CSDE1의 발현 수준 측정을 통한 삼중음성유방암 치료제 또는 전이 억제제 스크리닝 방법을 제공하는데 있다.In addition, another object of the present invention is to provide a method for screening triple negative breast cancer therapeutics or metastasis inhibitors by measuring the expression level of CSDE1.
또한, 본 발명의 또 다른 목적은 CSDE1 발현 또는 활성 억제제를 유효성분으로 포함하는, 시험관(in vitro) 내 삼중음성유방암세포에서의 RAC1 발현 억제용 시약 조성물을 제공하는데 있다.Another object of the present invention is to provide a reagent composition for inhibiting RAC1 expression in triple-negative breast cancer cells in vitro, comprising a CSDE1 expression or activity inhibitor as an active ingredient.
또한, 본 발명의 또 다른 목적은 시험관 내에서(in vitro) CSDE1 발현 또는 활성 억제제를 삼중음성유방암세포에 처리하는 단계를 포함하는 RAC1 발현 억제 방법을 제공하는데 있다.In addition, another object of the present invention is to provide a method for inhibiting RAC1 expression comprising treating triple negative breast cancer cells with an inhibitor of CSDE1 expression or activity in vitro.
상기 과제의 해결을 위하여, 본 발명은 CSDE1 또는 이를 코딩하는 유전자를 유효성분으로 포함하는 삼중음성유방암(Triple Negative Breast Cancer; TNBC) 진단 또는 전이 예후 예측용 바이오마커 조성물을 제공한다.In order to solve the above problems, the present invention provides a biomarker composition for diagnosing triple negative breast cancer (TNBC) or predicting metastasis prognosis, comprising CSDE1 or a gene encoding the same as an active ingredient.
또한, 본 발명은 CSDE1의 발현수준을 측정할 수 있는 제제를 유효성분으로 포함하는 삼중음성유방암 진단 또는 전이 예후 예측용 조성물을 제공한다.In addition, the present invention provides a composition for diagnosing triple-negative breast cancer or predicting the prognosis of metastasis, comprising an agent capable of measuring the expression level of CSDE1 as an active ingredient.
또한, 본 발명은 상기 조성물을 포함하는 삼중음성유방암 진단용 또는 전이 예후 예측용 키트를 제공한다.In addition, the present invention provides a kit for diagnosing triple-negative breast cancer or predicting the prognosis of metastasis comprising the composition.
또한, 본 발명은 (1) 환자에서 분리된 시료로부터 CSDE1 유전자의 mRNA 발현 수준 또는 CSDE1 단백질의 발현 수준을 측정하는 단계; (2) 상기 CSDE1 유전자의 mRNA 발현 수준 또는 CSDE1 단백질의 발현 수준을 대조군 시료와 비교하는 단계; 및 (3) 상기 CSDE1 유전자의 mRNA 발현 수준 또는 CSDE1 단백질의 발현 수준이 대조군 시료보다 높을 경우 삼중음성유방암이라고 판단하는 단계를 포함하는 삼중음성유방암 진단에 필요한 정보를 제공하는 방법을 제공한다.In addition, the present invention (1) measuring the mRNA expression level of the CSDE1 gene or the expression level of the CSDE1 protein from a sample isolated from the patient; (2) comparing the mRNA expression level of the CSDE1 gene or the expression level of the CSDE1 protein with a control sample; and (3) determining triple-negative breast cancer when the mRNA expression level of the CSDE1 gene or the CSDE1 protein expression level is higher than that of the control sample.
또한, 본 발명은 (1) 삼중음성유방암 환자에서 분리된 시료로부터 CSDE1 유전자의 mRNA 발현 수준 또는 CSDE1 단백질의 발현 수준을 측정하는 단계; (2) 상기 CSDE1 유전자의 mRNA 발현 수준 또는 CSDE1 단백질의 발현 수준을 대조군 시료와 비교하는 단계; 및 (3) 상기 CSDE1 유전자의 mRNA 발현 수준 또는 CSDE1 단백질의 발현 수준이 대조군 시료보다 높을 경우 삼중음성유방암의 전이 위험성이 높다고 판단하는 단계를 포함하는 삼중음성유방암 전이 예후 예측에 필요한 정보를 제공하는 방법을 제공한다.In addition, the present invention comprises the steps of (1) measuring the mRNA expression level of the CSDE1 gene or the expression level of the CSDE1 protein from a sample isolated from a triple-negative breast cancer patient; (2) comparing the mRNA expression level of the CSDE1 gene or the expression level of the CSDE1 protein with a control sample; and (3) determining that the risk of triple-negative breast cancer metastasis is high when the mRNA expression level of the CSDE1 gene or the CSDE1 protein expression level is higher than that of a control sample. provides
또한, (1) 삼중음성유방암 환자에서 분리된 시료로부터 CSDE1 유전자 및 RAC1 유전자의 mRNA 발현 수준 또는 CSDE1 단백질 및 RAC1 단백질의 발현 수준을 측정하는 단계; (2) 상기 CSDE1 유전자 및 RAC1 유전자의 mRNA 발현 수준 또는 CSDE1 단백질 및 RAC1 단백질의 발현 수준을 대조군 시료와 비교하는 단계; 및 (3) 상기 CSDE1 유전자 및 RAC1 유전자의 mRNA 발현 수준 또는 CSDE1 단백질 및 RAC1 단백질의 발현 수준이 대조군 시료보다 높을 경우 삼중음성유방암의 전이 위험성이 높다고 판단하는 단계를 포함하는 삼중음성유방암 전이 예후 예측에 필요한 정보를 제공하는 방법을 제공한다.In addition, (1) measuring the mRNA expression level of CSDE1 gene and RAC1 gene or the expression level of CSDE1 protein and RAC1 protein from a sample isolated from a triple-negative breast cancer patient; (2) comparing the mRNA expression level of the CSDE1 gene and the RAC1 gene or the expression level of the CSDE1 protein and the RAC1 protein with a control sample; and (3) determining that the risk of metastasis of triple-negative breast cancer is high when the mRNA expression levels of the CSDE1 gene and the RAC1 gene or the expression levels of the CSDE1 protein and the RAC1 protein are higher than those in the control sample. Provides a way to provide the necessary information.
또한, 본 발명은 CSDE1 발현 또는 활성 억제제를 유효성분으로 포함하는 삼중음성유방암 예방, 치료 또는 전이 억제용 약학조성물을 제공한다.In addition, the present invention provides a pharmaceutical composition for preventing, treating, or inhibiting metastasis of triple-negative breast cancer comprising a CSDE1 expression or activity inhibitor as an active ingredient.
또한, 본 발명은 (1) 삼중음성유방암 세포에 시험물질을 접촉시키는 단계; (2) 상기 시험물질을 접촉한 삼중음성유방암 세포에서 CSDE1의 발현 또는 활성 정도를 측정하는 단계; 및 (3) 대조군 시료와 비교하여 상기 CSDE1의 발현 또는 활성 정도가 감소한 시험물질을 선별하는 단계를 포함하는 삼중음성유방암 치료제 스크리닝 방법을 제공한다.In addition, the present invention comprises the steps of (1) contacting a test substance to triple-negative breast cancer cells; (2) measuring the expression or activity level of CSDE1 in triple-negative breast cancer cells contacted with the test substance; and (3) selecting a test substance having a reduced expression or activity level of the CSDE1 compared to a control sample.
또한, 본 발명은 (1) 삼중음성유방암 세포에 시험물질을 접촉시키는 단계; (2) 상기 시험물질을 접촉한 삼중음성유방암 세포에서 CSDE1의 발현 또는 활성 정도를 측정하는 단계; 및 (3) 대조군 시료와 비교하여 상기 CSDE1의 발현 또는 활성 정도가 감소한 시험물질을 선별하는 단계를 포함하는 삼중음성유방암 전이 억제제 스크리닝 방법을 제공한다.In addition, the present invention comprises the steps of (1) contacting a test substance to triple-negative breast cancer cells; (2) measuring the expression or activity level of CSDE1 in triple-negative breast cancer cells contacted with the test substance; and (3) selecting a test substance having a reduced expression or activity of the CSDE1 compared to a control sample.
또한, 본 발명은 (1) 삼중음성유방암 세포에 시험물질을 접촉시키는 단계; (2) 상기 시험물질을 접촉한 삼중음성유방암 세포에서 CSDE1 및 RAC1의 발현 또는 활성 정도를 측정하는 단계; 및 (3) 대조군 시료와 비교하여 상기 CSDE1 및 RAC1의 발현 또는 활성 정도가 감소한 시험물질을 선별하는 단계를 포함하는 삼중음성유방암 전이 억제제 스크리닝 방법을 제공한다.In addition, the present invention comprises the steps of (1) contacting a test substance to triple-negative breast cancer cells; (2) measuring the expression or activity levels of CSDE1 and RAC1 in triple-negative breast cancer cells contacted with the test substance; and (3) providing a screening method for triple-negative breast cancer metastasis inhibitors comprising the step of selecting a test substance in which the expression or activity of CSDE1 and RAC1 is reduced compared to a control sample.
또한, 본 발명은 CSDE1 발현 또는 활성 억제제를 유효성분으로 포함하는, 시험관(in vitro) 내 삼중음성유방암세포에서의 RAC1 발현 억제용 시약 조성물을 제공한다.In addition, the present invention provides a reagent composition for inhibiting RAC1 expression in triple-negative breast cancer cells in vitro, comprising a CSDE1 expression or activity inhibitor as an active ingredient.
또한, 본 발명은 시험관 내에서(in vitro) CSDE1 발현 또는 활성 억제제를 삼중음성유방암세포에 처리하는 단계를 포함하는 RAC1 발현 억제 방법을 제공한다.In addition, the present invention provides a method for inhibiting RAC1 expression comprising treating triple-negative breast cancer cells with an inhibitor of CSDE1 expression or activity in vitro.
본 발명은 CSDE1을 유효성분으로 포함하는 삼중음성유방암 전이 예측용 바이오마커 조성물에 관한 것으로서, CSDE1 유전자가 삼중음성유방암에서 발현이 높은 것으로 나타났고, 이는 침윤 및 전이와 관련된 유전자인 Rac1의 전사를 조절함을 확인하였다. 즉, 본 발명은 삼중음성유방암의 CSDE1의 발현을 기반으로, 암 전이와 관련된 Rac1 유전자의 전사 조절 메커니즘에 대해 제시하고 있다. 이를 이용하여, 본 발명은 삼중음성유방암의 진행 및 전이 예측을 위한 바이오마커로 유용하게 활용될 수 있다. 또한, 본 발명의 바이오마커를 이용하여 불필요한 치료를 줄이고 환자 맞춤형으로 치료전략을 수립할 수 있으며, 표적치료를 할 수 있는 항암제가 없는 삼중음성유방암 환자에게 맞는 적절한 치료를 통해 치료기간 및 치료비를 절감할 수 있을 것으로 기대된다.The present invention relates to a biomarker composition for predicting metastasis of triple-negative breast cancer comprising CSDE1 as an active ingredient. It was confirmed that That is, the present invention suggests a transcriptional regulatory mechanism of the Rac1 gene associated with cancer metastasis based on the expression of CSDE1 in triple-negative breast cancer. Using this, the present invention can be usefully utilized as a biomarker for predicting progression and metastasis of triple-negative breast cancer. In addition, using the biomarker of the present invention, it is possible to reduce unnecessary treatment and establish a treatment strategy tailored to the patient, and to reduce the treatment period and treatment cost through appropriate treatment suitable for triple-negative breast cancer patients without anticancer drugs capable of targeted treatment. Expect to be able to do it.
도 1은 삼중음성유방암 (triple negative breast cancer, TNBC) 모델에서 CSDE1 발현 증가 확인 및 전이/예후 예측 바이오마커로써 활용 가능성 검증한 결과를 나타낸다.Figure 1 shows the results of confirming the increase in CSDE1 expression in a triple negative breast cancer (TNBC) model and verifying the possibility of using it as a metastasis/prognosis predictive biomarker.
도 2는 CSDE1 발현 억제(siRNA, knock-out)를 통한 삼중음성유방암 세포주의 항암 효과 확인한 결과를 나타낸다.Figure 2 shows the results of confirming the anticancer effect of triple-negative breast cancer cell lines through CSDE1 expression inhibition (siRNA, knock-out).
도 3은 CSDE1 발현 기반 암 전이 유전자인 RAC1 발굴한 결과를 나타낸다.3 shows the results of discovering RAC1, a cancer metastasis gene based on CSDE1 expression.
도 4는 CSDE1에 의한 RAC1 전사 조절 메커니즘을 제시한 결과를 나타낸다.Figure 4 shows the results suggesting the mechanism of RAC1 transcriptional regulation by CSDE1.
도 5는 CSDE1, RAC1 발현 증가에 따른 삼중음성유방암 생존율 감소를 확인한 결과를 나타낸다.Figure 5 shows the results of confirming the decrease in triple-negative breast cancer survival rate according to the increased expression of CSDE1 and RAC1.
본 발명은 CSDE1 또는 이를 코딩하는 유전자를 유효성분으로 포함하는 삼중음성유방암(Triple Negative Breast Cancer; TNBC) 진단용 바이오마커 조성물을 제공한다. The present invention provides a biomarker composition for diagnosis of triple negative breast cancer (TNBC) comprising CSDE1 or a gene encoding the same as an active ingredient.
또한, 본 발명은 CSDE1 또는 이를 코딩하는 유전자를 유효성분으로 포함하는 삼중음성유방암 전이 예후 예측용 바이오마커 조성물을 제공한다. 바람직하게는, 상기 바이오마커 조성물은 RAC1 또는 이를 코딩하는 유전자를 더 포함할 수 있으나, 이에 한정되는 것은 아니다.In addition, the present invention provides a biomarker composition for predicting the prognosis of triple-negative breast cancer metastasis comprising CSDE1 or a gene encoding the same as an active ingredient. Preferably, the biomarker composition may further include RAC1 or a gene encoding the same, but is not limited thereto.
본 발명의 "CSDE1(cold shock domain containing E1)"은 NCBI accession no. NM_001007553.3, NM_007158.6, NM_001130523.3, NM_001242891.2, NM_001242892.2 또는 NM_001242893.2 일 수 있으나, 이에 한정되는 것은 아니다."CSDE1 (cold shock domain containing E1)" of the present invention is NCBI accession no. It may be NM_001007553.3, NM_007158.6, NM_001130523.3, NM_001242891.2, NM_001242892.2 or NM_001242893.2, but is not limited thereto.
본 발명의 "RAC1(Ras-related C3 botulinum toxin substrate 1)"은 NCBI accession no. NM_006908.5 또는 NM_018890.4 일 수 있으나, 이에 한정되는 것은 아니다."RAC1 (Ras-related C3 botulinum toxin substrate 1)" of the present invention is NCBI accession no. It may be NM_006908.5 or NM_018890.4, but is not limited thereto.
본 명세서에서 용어 “진단”은 특정 질병 또는 질환에 대한 한 객체의 감수성(susceptibility)을 판정하는 것, 한 객체가 특정 질병 또는 질환을 현재 가지고 있는지 여부를 판정하는 것, 특정 질병 또는 질환에 걸린 한 객체의 예후(prognosis)를 판정하는 것, 또는 테라메트릭스(therametrics)(예컨대, 치료 효능에 대한 정보를 제공하기 위하여 객체의 상태를 모니터링하는 것)을 포함한다.As used herein, the term "diagnosis" refers to determining a subject's susceptibility to a specific disease or disorder, determining whether a subject currently has a specific disease or disorder, as long as he or she has a specific disease or disorder. determining a subject's prognosis, or therametrics (eg, monitoring a subject's condition to provide information about treatment efficacy).
본 명세서 용어 “예후(prognosis)”는 질병을 진단하여 판단된 장래의 증세 또는 경과에 대한 전망을 말한다. 암 환자에 있어서 예후는 통상적으로 암 발병 또는 외과적 시술 후 일정기간 내의 전이 여부 또는 생존기간을 뜻한다. 예후의 예측은 특히 삼중음성유방암 환자의 화학치료 여부를 비롯하여 향후 삼중음성유방암 치료의 방향에 대한 단서를 제시하므로 매우 중요한 임상적 과제이다.The term "prognosis" in this specification refers to the outlook for future symptoms or progress determined by diagnosing a disease. In cancer patients, prognosis usually means metastasis or survival period within a certain period after cancer onset or surgical procedure. Prediction of prognosis is a very important clinical task because it provides clues to the direction of future triple-negative breast cancer treatment, including whether triple-negative breast cancer patients will undergo chemotherapy.
본 명세서에서 용어 “전이(metastasis)”는 어떤 종양이 그 원발 부위에서 여러 경로를 따라 다른 신체의 부위에 이식되어 그곳에 정착 및 증식하는 상태를 말한다. 암의 전이 여부는 해당 암의 고유의 특성에 의하여 결정될 뿐만 아니라 암의 예후 결정에 있어서 가장 중요한 단서가 되는 사건이므로, 암 환자의 생존과 관련된 가장 중요한 임상정보로 다루어진다. As used herein, the term "metastasis" refers to a state in which a certain tumor is transplanted from its primary site to another body part along various routes and settles and proliferates there. Metastasis of cancer is not only determined by the unique characteristics of the cancer, but also an event that is the most important clue in determining the prognosis of cancer, so it is treated as the most important clinical information related to the survival of cancer patients.
또한, 본 발명은 CSDE1의 발현수준을 측정할 수 있는 제제를 유효성분으로 포함하는 삼중음성유방암 진단용 조성물을 제공한다. In addition, the present invention provides a composition for diagnosis of triple-negative breast cancer comprising an agent capable of measuring the expression level of CSDE1 as an active ingredient.
또한, 본 발명은 CSDE1의 발현수준을 측정할 수 있는 제제를 유효성분으로 포함하는 삼중음성유방암 전이 예후 예측용 조성물을 제공한다. 바람직하게는, 상기 바이오마커 조성물은 RAC1 또는 이를 코딩하는 유전자를 더 포함할 수 있으나, 이에 한정되는 것은 아니다.In addition, the present invention provides a composition for predicting the prognosis of triple-negative breast cancer metastasis, comprising an agent capable of measuring the expression level of CSDE1 as an active ingredient. Preferably, the biomarker composition may further include RAC1 or a gene encoding the same, but is not limited thereto.
상세하게는, 상기 CSDE1 또는 RAC1의 발현수준을 측정할 수 있는 제제는 상기 CSDE1 또는 RAC1 유전자에 특이적으로 결합하는 프라이머 또는 프로브, 상기 CSDE1 또는 RAC1 단백질에 특이적으로 결합하는 항체, 펩타이드, 앱타머 또는 화합물일 수 있으나, 이에 한정되는 것은 아니다. Specifically, the agent capable of measuring the expression level of the CSDE1 or RAC1 is a primer or probe that specifically binds to the CSDE1 or RAC1 gene, an antibody, peptide, or aptamer that specifically binds to the CSDE1 or RAC1 protein Or it may be a compound, but is not limited thereto.
또한, 본 발명은 상기 조성물을 포함하는 삼중음성유방암 진단용 키트를 제공한다.In addition, the present invention provides a kit for diagnosing triple-negative breast cancer comprising the composition.
또한, 본 발명은 상기 조성물을 포함하는 삼중음성유방암 전이 예후 예측용 키트를 제공한다.In addition, the present invention provides a kit for predicting the prognosis of triple-negative breast cancer metastasis comprising the composition.
본 명세서에서 용어 "프라이머"는 짧은 자유 3-말단 수산화기(free 3' hydroxyl group)를 가지는 핵산 서열로 상보적인 템플레이트(template)와 염기쌍을 형성할 수 있고 템플레이트 가닥 복사를 위한 시작 지점으로서 작용하는 짧은 핵산 서열을 말한다. 프라이머는 적절한 완충용액 및 온도에서 중합반응을 위한 시약(즉, DNA 폴리머라제 또는 역전사효소) 및 상이한 4 가지의 뉴클레오사이드 트리포스페이트의 존재하에서 DNA 합성을 개시할 수 있다. PCR 조건, 센스 및 안티센스 프라이머의 길이는 당업계에 공지된 기술에 따라 적절히 선택될 수 있다.As used herein, the term "primer" refers to a nucleic acid sequence having a short free 3' hydroxyl group, capable of base pairing with a complementary template, and serving as a starting point for template strand copying. refers to the sequence of nucleic acids. Primers can initiate DNA synthesis in the presence of a reagent for polymerization (ie, DNA polymerase or reverse transcriptase) and four different nucleoside triphosphates in an appropriate buffer and temperature. PCR conditions and lengths of sense and antisense primers can be appropriately selected according to techniques known in the art.
본 명세서에서 용어 "프로브"는 mRNA외 특이적으로 결합을 이룰 수 있는 짧게는 수 염기 내지 길게는 수백 염기에 해당하는 RNA 또는 DNA 등의 핵산 단편을 의미하며 라벨링되어 있어서 특정 mRNA의 존재 유무, 발현양을 확인할 수 있다. 프로브는 올리고뉴클레오타이드(oligonucleotide) 프로브, 단쇄 DNA(single strand DNA) 프로브, 이중쇄DNA(double strand DNA) 프로브, RNA 프로브 등의 형태로 제작될 수 있다. 적절한 프로브의 선택 및 혼성화 조건은 당해 기술 분야에 공지된 기술에 따라 적절히 선택할 수 있다.As used herein, the term "probe" refers to a nucleic acid fragment such as RNA or DNA corresponding to a few bases to several hundred bases in length that can specifically bind to mRNA and is labeled, so that the presence or absence of a specific mRNA, expression quantity can be checked. The probe may be manufactured in the form of an oligonucleotide probe, a single strand DNA probe, a double strand DNA probe, an RNA probe, or the like. Selection of an appropriate probe and hybridization conditions can be appropriately selected according to techniques known in the art.
본 명세서에서 용어 "항체"는 당해 기술분야에 공지된 용어로서 항원성 부위에 대하여 지시되는 특이적인 면역 글로불린을 의미한다. 본 발명에서의 항체는 본 발명의 LRP-1에 대해 특이적으로 결합하는 항체를 의미하며, 당해 기술분야의 통상적인 방법에 따라 항체를 제조할 수 있다. 상기 항체의 형태는 폴리클로날 항체 또는 모노클로날 항체를 포함하며, 모든 면역글로불린 항체가 포함된다. 상기 항체는 2개의 전체 길이의 경쇄 및 2 개의 전체 길이의 중쇄를 갖는 완전한 형태를 의미한다. 또한, 상기 항체는 인간화 항체 등의 특수 항체도 포함된다.As used herein, the term "antibody" is a term known in the art and refers to a specific immunoglobulin directed against an antigenic site. The antibody in the present invention refers to an antibody that specifically binds to LRP-1 of the present invention, and the antibody can be prepared according to a conventional method in the art. The type of antibody includes polyclonal antibodies or monoclonal antibodies, and all immunoglobulin antibodies are included. The antibody is meant in its complete form with two full-length light chains and two full-length heavy chains. In addition, the antibody also includes special antibodies such as humanized antibodies.
또한, 본 발명의 키트는 마커 성분에 특이적으로 결합하는 항체, 기질과의 반응에 의해서 발색하는 표지체가 접합된 2차 항체 접합체(conjugate), 상기 표지체와 발색 반응할 발색 기질 용액, 세척액 및 효소반응 정지용액 등을 포함할 수 있으며, 사용되는 시약 성분을 포함하는 다수의 별도 패키징 또는 컴파트먼트로 제작될 수 있다.In addition, the kit of the present invention is an antibody that specifically binds to a marker component, a secondary antibody conjugate conjugated with a label that develops color by reaction with the substrate, a color-developing substrate solution that will react with the label, a washing solution, and It may contain an enzyme reaction stop solution, etc., and may be manufactured in a number of separate packaging or compartments containing reagent components to be used.
본 명세서에서 용어 "펩타이드"는 표적 물질에 대한 결합력 높은 장점이 있으며, 열/화학 처리시에도 변성이 일어나지 않는다. 또한 분자 크기가 작기 때문에 다른 단백질에 붙여서 융합 단백질로의 이용이 가능하다. 구체적으로 고분자 단백질 체인에 붙여서 이용이 가능하므로 진단 키트 및 약물전달 물질로 이용될 수 있다. As used herein, the term "peptide" has the advantage of having high binding ability to a target substance, and does not undergo denaturation even during heat/chemical treatment. In addition, because of its small molecular size, it can be attached to other proteins and used as a fusion protein. Specifically, since it can be used by attaching it to a polymer protein chain, it can be used as a diagnostic kit and a drug delivery material.
본 명세서에서 용어 "앱타머(aptamer)"란, 그 자체로 안정된 삼차 구조를 가지면서 표적 분자에 높은 친화성과 특이성으로 결합할 수 있는 특징을 가진 특별한 종류의 단일가닥 핵산(DNA, RNA 또는 변형핵산)으로 구성된 폴리뉴클레오티드의 일종을 의미한다. 상술한 바와 같이, 앱타머는 항체와 동일하게 항원성 물질에 특이적으로 결합할 수 있으면서도, 단백질보다 안정성이 높고, 구조가 간단하며, 합성이 용이한 폴리뉴클레오티드로 구성되어 있으므로, 항체를 대체하여 사용될 수 있다.As used herein, the term "aptamer" refers to a special kind of single-stranded nucleic acid (DNA, RNA or modified nucleic acid) having a stable tertiary structure and being able to bind to a target molecule with high affinity and specificity. ) means a type of polynucleotide composed of As described above, aptamers can specifically bind to antigenic substances in the same way as antibodies, but are more stable than proteins, have a simple structure, and are composed of polynucleotides that are easy to synthesize, so they can be used instead of antibodies. can
또한, 본 발명은 (1) 환자에서 분리된 시료로부터 CSDE1 유전자의 mRNA 발현 수준 또는 CSDE1 단백질의 발현 수준을 측정하는 단계; (2) 상기 CSDE1 유전자의 mRNA 발현 수준 또는 CSDE1 단백질의 발현 수준을 대조군 시료와 비교하는 단계; 및 (3) 상기 CSDE1 유전자의 mRNA 발현 수준 또는 CSDE1 단백질의 발현 수준이 대조군 시료보다 높을 경우 삼중음성유방암이라고 판단하는 단계를 포함하는 삼중음성유방암 진단에 필요한 정보를 제공하는 방법을 제공한다.In addition, the present invention (1) measuring the mRNA expression level of the CSDE1 gene or the expression level of the CSDE1 protein from a sample isolated from the patient; (2) comparing the mRNA expression level of the CSDE1 gene or the expression level of the CSDE1 protein with a control sample; and (3) determining triple-negative breast cancer when the mRNA expression level of the CSDE1 gene or the CSDE1 protein expression level is higher than that of the control sample.
또한, 본 발명은 (1) 삼중음성유방암 환자에서 분리된 시료로부터 CSDE1 유전자의 mRNA 발현 수준 또는 CSDE1 단백질의 발현 수준을 측정하는 단계; (2) 상기 CSDE1 유전자의 mRNA 발현 수준 또는 CSDE1 단백질의 발현 수준을 대조군 시료와 비교하는 단계; 및 (3) 상기 CSDE1 유전자의 mRNA 발현 수준 또는 CSDE1 단백질의 발현 수준이 대조군 시료보다 높을 경우 삼중음성유방암의 전이 위험성이 높다고 판단하는 단계를 포함하는 삼중음성유방암 전이 예후 예측에 필요한 정보를 제공하는 방법을 제공한다.In addition, the present invention comprises the steps of (1) measuring the mRNA expression level of the CSDE1 gene or the expression level of the CSDE1 protein from a sample isolated from a triple-negative breast cancer patient; (2) comparing the mRNA expression level of the CSDE1 gene or the expression level of the CSDE1 protein with a control sample; and (3) determining that the risk of triple-negative breast cancer metastasis is high when the mRNA expression level of the CSDE1 gene or the CSDE1 protein expression level is higher than that of a control sample. provides
또한, 본 발명은 (1) 삼중음성유방암 환자에서 분리된 시료로부터 CSDE1 유전자 및 RAC1 유전자의 mRNA 발현 수준 또는 CSDE1 단백질 및 RAC1 단백질의 발현 수준을 측정하는 단계; (2) 상기 CSDE1 유전자 및 RAC1 유전자의 mRNA 발현 수준 또는 CSDE1 단백질 및 RAC1 단백질의 발현 수준을 대조군 시료와 비교하는 단계; 및 (3) 상기 CSDE1 유전자 및 RAC1 유전자의 mRNA 발현 수준 또는 CSDE1 단백질 및 RAC1 단백질의 발현 수준이 대조군 시료보다 높을 경우 삼중음성유방암의 전이 위험성이 높다고 판단하는 단계를 포함하는 삼중음성유방암 전이 예후 예측에 필요한 정보를 제공하는 방법을 제공한다.In addition, the present invention (1) measuring the mRNA expression level of the CSDE1 gene and RAC1 gene or the expression level of the CSDE1 protein and RAC1 protein from a sample isolated from a triple-negative breast cancer patient; (2) comparing the mRNA expression level of the CSDE1 gene and the RAC1 gene or the expression level of the CSDE1 protein and the RAC1 protein with a control sample; and (3) determining that the risk of metastasis of triple-negative breast cancer is high when the mRNA expression levels of the CSDE1 gene and the RAC1 gene or the expression levels of the CSDE1 protein and the RAC1 protein are higher than those in the control sample. Provides a way to provide the necessary information.
상세하게는, 상기 mRNA 발현 수준을 측정하는 방법은 RT-PCR, 경쟁적 RT-PCR(Competitive RT-PCR), 실시간 RT-PCR (Real-time RT-PCR), RNase 보호 분석법(RPA; RNase protection assay), 노던 블랏팅 (Northern blotting) 및 DNA 칩을 이용하지만, 이에 한정되는 것은 아니다.Specifically, the method for measuring the mRNA expression level is RT-PCR, competitive RT-PCR (Competitive RT-PCR), real-time RT-PCR (Real-time RT-PCR), RNase protection assay (RPA; RNase protection assay ), Northern blotting, and DNA chips, but are not limited thereto.
상세하게는, 상기 단백질 발현 수준을 측정하는 방법은 웨스턴 블랏, ELISA(enzyme linked immunosorbent asay), 방사선면역분석(Radioimmunoassay; RIA), 방사면역확산법(radioimmunodiffusion), 오우크테로니(Ouchterlony) 면역 확산법, 로케이트(rocket) 면역전기영동, 조직면역염색, 면역침전 분석법(Immunoprecipitation assay), 보체고정분석법 (Complement Fixation Assay), FACS 및 단백질 칩을 이용하지만, 이에 한정되는 것은 아니다.Specifically, methods for measuring the protein expression level include Western blot, enzyme linked immunosorbent assay (ELISA), radioimmunoassay (RIA), radioimmunodiffusion, Ouchterlony immunodiffusion method, Rocket immunoelectrophoresis, tissue immunostaining, immunoprecipitation assay, complement fixation assay, FACS, and protein chip are used, but are not limited thereto.
본 명세서에서 용어 "환자에서 분리된 시료"란 바이오마커인 상기 CSDE1 또는 RAC1의 발현 수준에 있어서 대조군과 차이가 나는 조직, 세포, 전혈, 혈청, 혈장, 타액, 객담, 뇌척수액, 또는 뇨와 같은 시료를 포함하지만, 이에 한정되는 것은 아니다. As used herein, the term "sample isolated from a patient" means a sample such as tissue, cell, whole blood, serum, plasma, saliva, sputum, cerebrospinal fluid, or urine that is different from the control group in terms of the expression level of the CSDE1 or RAC1, which is a biomarker. Including, but not limited to.
또한, 본 발명은 CSDE1 발현 또는 활성 억제제를 유효성분으로 포함하는 삼중음성유방암 예방 또는 치료용 약학조성물을 제공한다. In addition, the present invention provides a pharmaceutical composition for preventing or treating triple-negative breast cancer comprising a CSDE1 expression or activity inhibitor as an active ingredient.
또한, 본 발명은 CSDE1 발현 또는 활성 억제제를 유효성분으로 포함하는 삼중음성유방암 전이 억제용 약학조성물을 제공한다. 바람직하게는, 상기 약학조성물은 RAC1 발현을 억제할 수 있으나, 이에 한정되는 것은 아니다.In addition, the present invention provides a pharmaceutical composition for inhibiting metastasis of triple-negative breast cancer comprising a CSDE1 expression or activity inhibitor as an active ingredient. Preferably, the pharmaceutical composition may inhibit RAC1 expression, but is not limited thereto.
상세하게는, 상기 CSDE1 단백질 발현 억제제는 CSDE1 유전자의 mRNA에 상보적으로 결합하는 안티센스 뉴클레오타이드, 작은 간섭 RNA(small interfering RNA; siRNA) 또는 짧은 헤어핀 RNA(short hairpin RNA; shRNA)일 수 있고, 상기 CSDE1 단백질 활성 억제제는 CSDE1 단백질에 특이적으로 결합하는 화합물, 펩티드, 펩티드 미메틱스, 앱타머, 항체 또는 천연물일 수 있으나, 이에 한정되는 것은 아니다.Specifically, the CSDE1 protein expression inhibitor may be an antisense nucleotide, small interfering RNA (siRNA) or short hairpin RNA (shRNA) that binds complementarily to the mRNA of the CSDE1 gene, and the CSDE1 Protein activity inhibitors may be compounds, peptides, peptide mimetics, aptamers, antibodies, or natural products that specifically bind to the CSDE1 protein, but are not limited thereto.
본 발명의 약학 조성물은 화학물질, 뉴클레오타이드, 안티센스, siRNA 올리고뉴클레오타이드 및 천연물 추출물을 유효성분으로 포함할 수 있다. 본 발명의 약학 조성물 또는 복합 제제는 유효 성분 이외에 약제학적으로 적합하고 생리학적으로 허용되는 보조제를 사용하여 제조될 수 있으며, 상기 보조제로는 부형제, 붕해제, 감미제, 결합제, 피복제, 팽창제, 윤활제, 활택제 또는 향미제 등의 가용화제를 사용할 수 있다. 본 발명의 약학 조성물은 투여를 위해서 유효 성분 이외에 추가로 약제학적으로 허용 가능한 담체를 1 종 이상 포함하여 약학 조성물로 바람직하게 제제화할 수 있다. 액상 용액으로 제제화되는 조성물에 있어서 허용 가능한 약제학적 담체로는, 멸균 및 생체에 적합한 것으로서, 식염수, 멸균수, 링거액, 완충 식염수, 알부민 주사용액, 덱스트로즈 용액, 말토 덱스트린 용액, 글리세롤, 에탄올 및 이들 성분 중 1 성분 이상을 혼합하여 사용할 수 있으며, 필요에 따라 항산화제, 완충액, 정균제 등 다른 통상의 첨가제를 첨가할 수 있다. 또한 희석제, 분산제, 계면활성제, 결합제 및 윤활제를 부가적으로 첨가하여 수용액, 현탁액, 유탁액 등과 같은 주사용 제형, 환약, 캡슐, 과립 또는 정제로 제제화할 수 있다. The pharmaceutical composition of the present invention may include chemical substances, nucleotides, antisense, siRNA oligonucleotides, and natural extracts as active ingredients. The pharmaceutical composition or combined preparation of the present invention can be prepared using pharmaceutically suitable and physiologically acceptable adjuvants in addition to the active ingredients, and the adjuvants include excipients, disintegrants, sweeteners, binders, coating agents, expanding agents, and lubricants. , solubilizing agents such as lubricants or flavoring agents may be used. The pharmaceutical composition of the present invention may be preferably formulated as a pharmaceutical composition by including one or more pharmaceutically acceptable carriers in addition to the active ingredient for administration. In compositions formulated as liquid solutions, acceptable pharmaceutical carriers are sterile and biocompatible, and include saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol and One or more of these components may be mixed and used, and other conventional additives such as antioxidants, buffers, and bacteriostatic agents may be added if necessary. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to prepare formulations for injections such as aqueous solutions, suspensions, and emulsions, pills, capsules, granules, or tablets.
본 발명의 약학 조성물의 약제 제제 형태는 과립제, 산제, 피복정, 정제, 캡슐제, 좌제, 시럽, 즙, 현탁제, 유제, 점적제 또는 주사 가능한 액제 및 활성 화합물의 서방출형 제제 등이 될 수 있다. 본 발명의 약학 조성물은 정맥내, 동맥내, 복강내, 근육내, 동맥내, 복강내, 흉골내, 경피, 비측내, 흡입, 국소, 직장, 경구, 안구내 또는 피내 경로를 통해 통상적인 방식으로 투여할 수 있다. 본 발명의 약학 조성물의 유효성분의 유효량은 질환의 예방 또는 치료 요구되는 양을 의미한다. 따라서, 질환의 종류, 질환의 중증도, 조성물에 함유된 유효 성분 및 다른 성분의 종류 및 함량, 제형의 종류 및 환자의 연령, 체중, 일반 건강 상태, 성별 및 식이, 투여 시간, 투여 경로 및 조성물의 분비율, 치료 기간, 동시 사용되는 약물을 비롯한 다양한 인자에 따라 조절될 수 있다. 이에 제한되는 것은 아니나, 예컨대, 성인의 경우, 1일 1회 내지 수회 투여시, 본 발명의 조성물은 1일 1회 내지 수회 투여시, 화합물일 경우 0.1ng/kg~10g/kg, 폴리펩타이드, 단백질 또는 항체일 경우 0.1ng/kg~10g/kg, 안티센스 뉴클레오타이드, siRNA, shRNAi, miRNA일 경우 0.01ng/kg~10g/kg의 용량으로 투여할 수 있다. The pharmaceutical formulation form of the pharmaceutical composition of the present invention may be granules, powders, coated tablets, tablets, capsules, suppositories, syrups, juices, suspensions, emulsions, drops or injectable solutions, and sustained-release preparations of active compounds. can The pharmaceutical composition of the present invention can be administered in a conventional manner via intravenous, intraarterial, intraperitoneal, intramuscular, intraarterial, intraperitoneal, intrasternal, transdermal, intranasal, inhalational, topical, rectal, oral, intraocular or intradermal routes. can be administered with An effective amount of the active ingredient of the pharmaceutical composition of the present invention means an amount required for preventing or treating a disease. Therefore, the type of disease, the severity of the disease, the type and amount of the active ingredient and other ingredients contained in the composition, the type of formulation and the patient's age, weight, general health condition, sex and diet, administration time, administration route and composition It can be controlled by various factors including secretion rate, duration of treatment, and drugs used concurrently. Although not limited thereto, for example, in the case of adults, when administered once to several times a day, the composition of the present invention is administered once to several times a day, in the case of a compound, 0.1 ng / kg to 10 g / kg, polypeptide, It can be administered at a dose of 0.1 ng/kg to 10 g/kg for proteins or antibodies and 0.01 ng/kg to 10 g/kg for antisense nucleotides, siRNA, shRNAi, and miRNA.
또한, 본 발명은 (1) 삼중음성유방암 세포에 시험물질을 접촉시키는 단계; (2) 상기 시험물질을 접촉한 삼중음성유방암 세포에서 CSDE1의 발현 또는 활성 정도를 측정하는 단계; 및 (3) 대조군 시료와 비교하여 상기 CSDE1의 발현 또는 활성 정도가 감소한 시험물질을 선별하는 단계를 포함하는 삼중음성유방암 치료제 스크리닝 방법을 제공한다. In addition, the present invention comprises the steps of (1) contacting a test substance to triple-negative breast cancer cells; (2) measuring the expression or activity level of CSDE1 in triple-negative breast cancer cells contacted with the test substance; and (3) selecting a test substance having a reduced expression or activity level of the CSDE1 compared to a control sample.
또한, 본 발명은 (1) 삼중음성유방암 세포에 시험물질을 접촉시키는 단계; (2) 상기 시험물질을 접촉한 삼중음성유방암 세포에서 CSDE1의 발현 또는 활성 정도를 측정하는 단계; 및 (3) 대조군 시료와 비교하여 상기 CSDE1의 발현 또는 활성 정도가 감소한 시험물질을 선별하는 단계를 포함하는 삼중음성유방암 전이 억제제 스크리닝 방법을 제공한다.In addition, the present invention comprises the steps of (1) contacting a test substance to triple-negative breast cancer cells; (2) measuring the expression or activity level of CSDE1 in triple-negative breast cancer cells contacted with the test substance; and (3) selecting a test substance having a reduced expression or activity of the CSDE1 compared to a control sample.
또한, 본 발명은 (1) 삼중음성유방암 세포에 시험물질을 접촉시키는 단계; (2) 상기 시험물질을 접촉한 삼중음성유방암 세포에서 CSDE1 및 RAC1의 발현 또는 활성 정도를 측정하는 단계; 및 (3) 대조군 시료와 비교하여 상기 CSDE1 및 RAC1의 발현 또는 활성 정도가 감소한 시험물질을 선별하는 단계를 포함하는 삼중음성유방암 전이 억제제 스크리닝 방법을 제공한다.In addition, the present invention comprises the steps of (1) contacting a test substance to triple-negative breast cancer cells; (2) measuring the expression or activity levels of CSDE1 and RAC1 in triple-negative breast cancer cells contacted with the test substance; and (3) providing a screening method for triple-negative breast cancer metastasis inhibitors comprising the step of selecting a test substance in which the expression or activity of CSDE1 and RAC1 is reduced compared to a control sample.
본 발명의 스크리닝 방법을 언급하면서 사용되는 용어 "시험물질"은 유전자의 발현량에 영향을 미치거나, 단백질의 발현 또는 활성에 영향을 미치는지 여부를 검사하기 위하여 스크리닝에서 이용되는 미지의 후보 물질을 의미한다. 상기 시료는 화학물질, 뉴클레오타이드, 안티센스-RNA, siRNA(small interference RNA) 및 천연물 추출물을 포함하나, 이에 제한되는 것은 아니다. The term "test substance" used while referring to the screening method of the present invention refers to an unknown candidate substance used in screening to test whether it affects the expression level of a gene or affects the expression or activity of a protein. do. The samples include, but are not limited to, chemical substances, nucleotides, antisense-RNA, small interference RNA (siRNA), and natural product extracts.
또한, 본 발명은 CSDE1 발현 또는 활성 억제제를 유효성분으로 포함하는, 시험관(in vitro) 내 삼중음성유방암세포에서의 RAC1 발현 억제용 시약 조성물을 제공한다.In addition, the present invention provides a reagent composition for inhibiting RAC1 expression in triple-negative breast cancer cells in vitro, comprising a CSDE1 expression or activity inhibitor as an active ingredient.
또한, 본 발명은 시험관 내에서(in vitro) CSDE1 발현 또는 활성 억제제를 삼중음성유방암세포에 처리하는 단계를 포함하는 RAC1 발현 억제 방법을 제공한다.In addition, the present invention provides a method for inhibiting RAC1 expression comprising treating triple-negative breast cancer cells with an inhibitor of CSDE1 expression or activity in vitro.
이하, 본 발명의 이해를 돕기 위하여 실시예를 들어 상세하게 설명하기로 한다. 다만 하기의 실시예는 본 발명의 내용을 예시하는 것일 뿐 본 발명의 범위가 하기 실시예에 한정되는 것은 아니다. 본 발명의 실시예는 당업계에서 평균적인 지식을 가진 자에게 본 발명을 보다 완전하게 설명하기 위해 제공되는 것이다.Hereinafter, examples will be described in detail to aid understanding of the present invention. However, the following examples are merely illustrative of the contents of the present invention, but the scope of the present invention is not limited to the following examples. The embodiments of the present invention are provided to more completely explain the present invention to those skilled in the art.
<실험예><Experimental example>
하기의 실험예들은 본 발명에 따른 각각의 실시예에 공통적으로 적용되는 실험예를 제공하기 위한 것이다.The following experimental examples are intended to provide experimental examples commonly applied to each embodiment according to the present invention.
1) TNBC_TCGA 데이터 분석: cBioportal 과 TCGA breast cancer dataset 분석을 통해 유방암 subtype 특이적인 CSDE1 유전자 발현을 비교 분석하였다. 1) TNBC_TCGA data analysis: Through cBioportal and TCGA breast cancer dataset analysis, breast cancer subtype-specific CSDE1 gene expression was compared and analyzed.
2) qRT-PCR 분석: 특정 유전자의 발현 변화를 확인하기 위해 Nucleospin kit를 이용하여, mRNA를 추출하였다. 추출된 mRNA를 reverse transcriptase (RTase)를 이용하여 cDNA로 합성한 후, 특정 유전자에 대한 프라이머(primer)를 사용하여 qRT-PCR 분석을 진행하였다.2) qRT-PCR analysis: mRNA was extracted using Nucleospin kit to confirm the expression change of a specific gene. After synthesizing the extracted mRNA into cDNA using reverse transcriptase (RTase), qRT-PCR analysis was performed using a primer for a specific gene.
3) Western blot (단백질 발현 분석): 세포에서 단백질 추출을 위해 Nucleospin kit를 이용하였으며, SDS-PAGE gel에 단백질을 로딩(loading)하여 분자량(molecular weight) 순으로 진행하였다. SDS-PAGE gel의 단백질을 멤브레인(membrane)에 옮긴 후, 1차 항체, 2차 항체 순으로 반응시켰으며, ECL solution을 이용해 imaging 장비로 단백질 밴드(band)를 검출하였다.3) Western blot (protein expression analysis): Nucleospin kit was used to extract proteins from cells, and proteins were loaded on SDS-PAGE gel and proceeded in order of molecular weight. After transferring the protein from the SDS-PAGE gel to a membrane, the reaction was performed in the order of primary antibody and secondary antibody, and protein bands were detected by imaging equipment using ECL solution.
4) Cell transfection: Lipofectamine RNAiMax 와 FuGene 시약을 이용하여 siRNA와 플라스미드(plasmid)를 각각 형질감염(transfection) 시킨 후, 24시간 후에 다양한 in vitro assay 실험을 수행하였다.4) Cell transfection: After transfection of siRNA and plasmid using Lipofectamine RNAiMax and FuGene reagent, various in vitro assay experiments were performed 24 hours later.
5) Cell viability assay: 세포를 96-well에 동일한 세포수로 접종(seeding)하였다. 접종 후, 24시간 간격으로 CCK-8 (cell counting kit-8) 시약을 이용하여 ELISA 기기를 통해 세포 생존율(cell viability)을 측정하였다.5) Cell viability assay: Cells were seeded in 96-well with the same number of cells. After inoculation, cell viability was measured at 24-hour intervals using an ELISA device using CCK-8 (cell counting kit-8) reagent.
6) Transwell migration assay: Transwell insert를 24-well에 넣은 후, 무혈청(serum-free) 배지로 현탁(suspension) 된 세포를 insert에 접종하였고, insert membrane 아래에 10% FBS 배지를 넣어 준 후, 이동(migration) 된 세포를 crystal violet 시약 염색을 통해 관찰하였다.6) Transwell migration assay: After inserting Transwell insert into 24-well, cells suspended in serum-free medium were inoculated into the insert, and 10% FBS medium was placed under the insert membrane, Migrated cells were observed by staining with crystal violet reagent.
7) Transwell invasion assay: Matrigel로 코팅된 transwell insert를 24-well에 넣은 후, 무혈청(serum-free) 배지로 현탁(suspension) 된 세포를 insert에 접종하였고, insert membrane 아래에 10% FBS 배지를 넣어 준 후, 침습(invasion)한 세포를 crystal violet 시약 염색을 통해 관찰하였다.7) Transwell invasion assay: Matrigel-coated transwell insert was placed in 24-well, cells suspended in serum-free medium were inoculated into the insert, and 10% FBS medium was placed under the insert membrane. After putting in, the invaded cells were observed through crystal violet reagent staining.
8) colony formation assay: 6-well에 세포를 동일한 양으로 접종한 후, 약 7일 후에 crystal violet 염색을 통해 colony formation 차이를 관찰하였다.8) Colony formation assay: After inoculating the same amount of cells into 6-well, differences in colony formation were observed through crystal violet staining after about 7 days.
9) Immunoprecipitation (IP): 실험에 준비된 세포를 IP lysis buffer로 용해(lysis) 시킨 후, 표적 단백질(target protein)을 풀 다운(pull down) 할 수 있는 항체와 함께 반응시켰다. 항체를 풀 다운(pull down) 할 수 있는 Dynabead를 사용하여 단백질, 항체 복합체(complex)를 풀 다운(pull down) 한 후에, western blot을 수행하여, 특정 단백질 간의 결합 여부를 확인하였다. 9) Immunoprecipitation (IP): After the cells prepared for the experiment were lysed with IP lysis buffer, they were reacted with an antibody capable of pulling down the target protein. After pulling down proteins and antibody complexes using Dynabead capable of pulling down antibodies, Western blot was performed to confirm whether specific proteins were bound.
10) RNA-IP (immunoprecipitation): 세로를 1% 포름알데히드(formaldehyde)로 가교(cross-link) 시킨 후, lysis buffer로 현탁(suspension) 시켰다. 확보된 pre-cleared supernatant를 확인하고자 하는 1차 항체와 반응시킨 후, A/G magnetic bead를 사용하여 풀 다운(pull down) 시켰다. 확보된 pellet bead로부터 RNA를 확보한 후, qRT-PCR 분석을 통해 확인하였다. 10) RNA-IP (immunoprecipitation): Cells were cross-linked with 1% formaldehyde and suspended in lysis buffer. After reacting the obtained pre-cleared supernatant with the primary antibody to be identified, it was pulled down using an A/G magnetic bead. After securing RNA from the secured pellet beads, it was confirmed through qRT-PCR analysis.
<< 실시예Example 1> 삼중음성유방암(triple negative breast cancer, 1> triple negative breast cancer, TNBCTNBC ) 모델에서 CSDE1 발현 증가 확인 및 전이/예후 예측 바이오마커로써 활용 가능성 검증 ) Confirmation of CSDE1 expression increase in the model and verification of the possibility of use as a predictive biomarker for metastasis / prognosis
non-TNBC 세포주와 TNBC 세포주에서 CSDE1 단백질 발현을 western blot 실험을 통해 분석한 결과, TNBC 세포주에서 CSDE1 단백질 발현이 증가되어 있음을 확인하였다(도 1A). 유방암 환자의 subtype 별 CSDE1 mRNA 발현을 TCGA 데이터를 통해 분석 한 결과, basal-like (TNBC) 환자군에서 CSDE1 mRNA 발현이 특이적으로 증가되어 있음을 확인하였다(도 1B). 또한, TCGA 데이터 분석을 통해, 림프절 전이가 있는 TNBC 환자군에서 CSDE1 발현이 증가될수록 생존율이 감소하는 경향성을 확인하였다(도 1C). 이러한 결과는 CSDE1 발현이 삼중음성유방암 특이적으로 증가되어 있으며, CSDE1 발현 증가가 삼중음성유방암의 전이/예후 예측 바이오마커로 활용 될 수 있음을 제시하고 있다.As a result of analyzing CSDE1 protein expression in non-TNBC cell lines and TNBC cell lines through western blot experiments, it was confirmed that CSDE1 protein expression was increased in TNBC cell lines (FIG. 1A). As a result of analyzing CSDE1 mRNA expression by subtype of breast cancer patients through TCGA data, it was confirmed that CSDE1 mRNA expression was specifically increased in the basal-like (TNBC) patient group (Fig. 1B). In addition, through TCGA data analysis, it was confirmed that the survival rate decreased as CSDE1 expression increased in the TNBC patient group with lymph node metastasis (FIG. 1C). These results suggest that CSDE1 expression is specifically increased in triple-negative breast cancer, and that increased CSDE1 expression can be used as a predictive biomarker for metastasis/prognosis of triple-negative breast cancer.
<< 실시예Example 2> 2> CSDE1CSDE1 발현 억제( expression inhibition ( siRNAsiRNA , knock-out)를 통한 삼중음성유방암 세포주의 항암 효과 확인Confirmation of anticancer effect of triple negative breast cancer cell line through knock-out)
삼중음성유방암 모델에서 증가되어 있는 CSDE1 발현을 서로 다른 두 종류의 CSDE1 siRNA 형질감염(transfection)을 통하여 감소시킨 후, 세포 생존율(cell viability)을 측정하였다. 그 결과, CSDE1 siRNA가 형질감염(transfection) 된 TNBC 세포주에서 세포 생존율(cell viability)이 유의미하게 감소함을 확인하였음 (도 2A). 또한, CSDE1 발현이 knock-out 된 TNBC 세포주에서 transwell assay를 수행한 결과, 이동(migration), 침습(invasion) 억제 효과를 보였으며(도 2B), 콜로니 형성(colony formation) 능력도 감소됨을 확인하였다(도 2C). 이러한 결과는 TNBC 에서 증가되어 있는 CSDE1 발현을 감소시켰을 때, TNBC 세포주에 항암 효과가 있음을 검증한 결과이다.After reducing CSDE1 expression, which is increased in the triple-negative breast cancer model, through two different types of CSDE1 siRNA transfection, cell viability was measured. As a result, it was confirmed that cell viability was significantly reduced in the TNBC cell line transfected with CSDE1 siRNA (Fig. 2A). In addition, as a result of transwell assay performed on TNBC cell lines in which CSDE1 expression was knocked out, migration and invasion were inhibited (Fig. 2B), and colony formation ability was also reduced. It was confirmed (Fig. 2C). These results are the results of verifying that there is an anticancer effect in TNBC cell lines when CSDE1 expression, which is increased in TNBC, is reduced.
<실시예 3> CSDE1 발현 기반 암 전이 유전자인 RAC1 발굴<Example 3> Discovery of RAC1, a cancer metastasis gene based on CSDE1 expression
삼중음성유방암에서 증가되어 있는 CSDE1 발현이 암 전이 유전자인 RAC1 발현 조절에 영향을 미칠 수 있는지 검증하였다. 삼중음성유방암 세포주 (MDA-MB-231, Hs578T) 에서 CSDE1 발현을 siRNA 형질감염(transfection)을 통해 감소시켜주었을 때, RAC1의 mRNA, 단백질 발현 모두 감소됨을 확인하였다(도 3A 및 도 3B). 또한, CSDE1 유전자가 knock-out (KO) 된 TNBC 세포주에서도 RAC1 단백질 발현이 감소되어 있음을 확인하였다(도 3C). 추가로, 정상 세포주인 MCF10A와 HEK293T 세포에 CSDE1 발현을 플라스미드 형질감염(plasmid transfection)을 통해 증가시켜주었을 때, RAC1 발현이 증가함을 확인하였다(도 3D). 이러한 결과는 CSDE1이 암 전이 유전자인 RAC1 발현 조절에 직접적으로 관여하고 있음을 제시하고 있다.We verified whether CSDE1 expression, which is increased in triple-negative breast cancer, could affect the regulation of RAC1 expression, a cancer metastasis gene. When CSDE1 expression was reduced in triple-negative breast cancer cell lines (MDA-MB-231, Hs578T) through siRNA transfection, it was confirmed that both mRNA and protein expression of RAC1 were reduced (FIGS. 3A and 3B). In addition, it was confirmed that RAC1 protein expression was decreased in the TNBC cell line in which the CSDE1 gene was knocked out (KO) (FIG. 3C). Additionally, when CSDE1 expression was increased in normal cell lines MCF10A and HEK293T cells through plasmid transfection, it was confirmed that RAC1 expression increased (FIG. 3D). These results suggest that CSDE1 is directly involved in regulating the expression of RAC1, a cancer metastasis gene.
<실시예 4> CSDE1에 의한 RAC1 전사 조절 메커니즘 제시<Example 4> Presentation of RAC1 transcriptional regulation mechanism by CSDE1
CSDE1이 RAC1을 조절함에 있어 RAC1의 전사 조절 수준에서 어떠한 단계에 특이적으로 작용하는지 검증하였다. 우선 CSDE1과 RNA polymerase II p-CTD (ser2), RNA polymerase II p-CTD (ser5), RNA polymerase II p-CTD (ser7)과의 결합(interaction) 여부를 면역침전분석(immunoprecipitation; IP)을 통해 확인한 결과, CSDE1이 각각의 RNA pol II 와 모두 결합하고 있음을 확인하였다(도 4A). RNA pol II의 ser5, ser7 이 전사 조절 수준에서 개시(initiation), 연장(elongation) 마커로 사용되고 있으며, ser2는 종결(termination) 마커로 사용되고 있음을 고려하였을 때, CSDE1은 특정 유전자의 전사 조절에 있어서 개시(initiation), 연장(elongation), 종결(termination) 단계에 모두 관여할 수 있음을 제시하고 있다. 또한, 실제로 CSDE1이 RAC1을 조절함에 있어서, 어떠한 RNA pol II 와 결합하는지 RNA-IP를 통해 확인한 결과, CSDE1이 ser2, ser5, ser7과 모두 결합(interaction) 하면서 RAC1 조절에 관여할 수 있음을 검증하였다(도 4B). 이러한 결과를 통해, CSDE1이 RAC1의 전사 조절 수준의 개시(initiation), 연장(elongation), 종결(termination)에 관여할 수 있음을 제시하였다.In the regulation of RAC1 by CSDE1, it was verified which step in the level of transcription regulation of RAC1 specifically acts. First, the interaction between CSDE1 and RNA polymerase II p-CTD (ser2), RNA polymerase II p-CTD (ser5), and RNA polymerase II p-CTD (ser7) was examined through immunoprecipitation (IP). As a result of confirmation, it was confirmed that CSDE1 binds to each RNA pol II (FIG. 4A). Considering that ser5 and ser7 of RNA pol II are used as initiation and elongation markers at the level of transcriptional regulation, and ser2 is used as a termination marker, CSDE1 has a role in transcriptional regulation of specific genes. It suggests that it can be involved in all stages of initiation, elongation, and termination. In addition, as a result of confirming through RNA-IP which RNA pol II binds to which RNA pol II CSDE1 actually regulates RAC1, it was verified that CSDE1 can be involved in RAC1 regulation while interacting with all ser2, ser5, and ser7 (Fig. 4B). These results suggest that CSDE1 may be involved in the initiation, elongation, and termination of RAC1 transcriptional regulation.
<< 실시예Example 5> 5> CSDE1CSDE1 , , RAC1RAC1 발현 증가에 따른 삼중음성유방암 생존율 감소 확인 Confirmation of decrease in triple-negative breast cancer survival rate due to increased expression
CSDE1과 RAC1 유전자 발현 측면에서의 상관관계를 통해, CSDE1과 RAC1 발현이 삼중음성유방암 환자의 예후 측면에서 임상학적 의미를 갖는지 확인하였다. 그 결과, CSDE1과 RAC1 발현이 동시에 증가되어 있는 삼중음성유방암 환자에서 생존율이 감소함을 확인하였다(도 5). 이러한 결과를 종합해보았을 때, CSDE1과 RAC1을 삼중음성유방암의 진단/예후 예측에 활용 가능한 바이오마커로써 제시할 수 있었다.Through the correlation between CSDE1 and RAC1 gene expression, it was confirmed whether CSDE1 and RAC1 expression have clinical significance in terms of prognosis of triple-negative breast cancer patients. As a result, it was confirmed that the survival rate decreased in triple-negative breast cancer patients in which CSDE1 and RAC1 expressions were simultaneously increased (FIG. 5). Taken together, these results suggest that CSDE1 and RAC1 can be used as biomarkers for the diagnosis/prognosis of triple-negative breast cancer.
이상으로 본 발명의 특정한 부분을 상세히 기술하였는 바, 당업계의 통상의 지식을 가진 자에게 있어서, 이러한 구체적 기술은 단지 바람직한 실시예일 뿐이며, 이에 의해 본 발명의 범위가 제한되는 것이 아닌 점은 명백할 것이다. 따라서, 본 발명의 실질적인 범위는 첨부된 청구항들과 그것들의 등가물에 의하여 정의된다고 할 것이다. Having described specific parts of the present invention in detail above, it will be clear to those skilled in the art that these specific descriptions are merely preferred embodiments, and the scope of the present invention is not limited thereby. will be. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

Claims (20)

  1. CSDE1 또는 이를 코딩하는 유전자를 유효성분으로 포함하는 삼중음성유방암(Triple Negative Breast Cancer; TNBC) 진단용 바이오마커 조성물.A biomarker composition for diagnosis of triple negative breast cancer (TNBC) comprising CSDE1 or a gene encoding the same as an active ingredient.
  2. CSDE1 또는 이를 코딩하는 유전자를 유효성분으로 포함하는 삼중음성유방암 전이 예후 예측용 바이오마커 조성물.A biomarker composition for predicting the prognosis of triple-negative breast cancer metastasis comprising CSDE1 or a gene encoding the same as an active ingredient.
  3. 제2항에 있어서, 상기 바이오마커 조성물은 RAC1 또는 이를 코딩하는 유전자를 더 포함하는 것을 특징으로 하는 삼중음성유방암 전이 예후 예측용 바이오마커 조성물.The biomarker composition for predicting the prognosis of triple-negative breast cancer metastasis according to claim 2, wherein the biomarker composition further comprises RAC1 or a gene encoding the same.
  4. CSDE1의 발현수준을 측정할 수 있는 제제를 유효성분으로 포함하는 삼중음성유방암 진단용 조성물.A composition for diagnosis of triple-negative breast cancer comprising an agent capable of measuring the expression level of CSDE1 as an active ingredient.
  5. CSDE1의 발현수준을 측정할 수 있는 제제를 유효성분으로 포함하는 삼중음성유방암 전이 예후 예측용 조성물.A composition for predicting the prognosis of triple-negative breast cancer metastasis, comprising an agent capable of measuring the expression level of CSDE1 as an active ingredient.
  6. 제5항에 있어서, 상기 조성물은 RAC1의 발현수준을 측정할 수 있는 제제를 더 포함하는 것을 특징으로 하는 삼중음성유방암 전이 예후 예측용 조성물.The composition for predicting the prognosis of triple-negative breast cancer metastasis according to claim 5, wherein the composition further comprises an agent capable of measuring the expression level of RAC1.
  7. 제4항 내지 제6항 중 어느 한 항에 있어서, 상기 CSDE1 또는 RAC1의 발현 수준을 측정할 수 있는 제제는 상기 CSDE1 유전자 또는 RAC1 유전자에 특이적으로 결합하는 프라이머 또는 프로브, 상기 CSDE1 단백질 또는 RAC1 단백질에 특이적으로 결합하는 항체, 펩타이드, 앱타머 또는 화합물인 것을 특징으로 하는 조성물.The method according to any one of claims 4 to 6, wherein the agent capable of measuring the expression level of the CSDE1 or RAC1 is a primer or probe that specifically binds to the CSDE1 gene or the RAC1 gene, the CSDE1 protein or the RAC1 protein A composition characterized in that the antibody, peptide, aptamer or compound that specifically binds to.
  8. 제4항의 조성물을 포함하는 삼중음성유방암 진단용 키트.A kit for diagnosing triple-negative breast cancer comprising the composition of claim 4.
  9. 제5항 또는 제6항의 조성물을 포함하는 삼중음성유방암 전이 예후 예측용 키트.A kit for predicting the prognosis of triple-negative breast cancer metastasis comprising the composition of claim 5 or claim 6.
  10. (1) 환자에서 분리된 시료로부터 CSDE1 유전자의 mRNA 발현 수준 또는 CSDE1 단백질의 발현 수준을 측정하는 단계; (1) measuring the mRNA expression level of the CSDE1 gene or the expression level of the CSDE1 protein from a sample isolated from the patient;
    (2) 상기 CSDE1 유전자의 mRNA 발현 수준 또는 CSDE1 단백질의 발현 수준을 대조군 시료와 비교하는 단계; 및(2) comparing the mRNA expression level of the CSDE1 gene or the expression level of the CSDE1 protein with a control sample; and
    (3) 상기 CSDE1 유전자의 mRNA 발현 수준 또는 CSDE1 단백질의 발현 수준이 대조군 시료보다 높을 경우 삼중음성유방암이라고 판단하는 단계를 포함하는 삼중음성유방암 진단에 필요한 정보를 제공하는 방법.(3) A method for providing information necessary for diagnosing triple-negative breast cancer, including determining triple-negative breast cancer when the mRNA expression level of the CSDE1 gene or the CSDE1 protein expression level is higher than that of a control sample.
  11. (1) 삼중음성유방암 환자에서 분리된 시료로부터 CSDE1 유전자의 mRNA 발현 수준 또는 CSDE1 단백질의 발현 수준을 측정하는 단계; (1) measuring the mRNA expression level of the CSDE1 gene or the expression level of the CSDE1 protein from a sample isolated from a triple-negative breast cancer patient;
    (2) 상기 CSDE1 유전자의 mRNA 발현 수준 또는 CSDE1 단백질의 발현 수준을 대조군 시료와 비교하는 단계; 및(2) comparing the mRNA expression level of the CSDE1 gene or the expression level of the CSDE1 protein with a control sample; and
    (3) 상기 CSDE1 유전자의 mRNA 발현 수준 또는 CSDE1 단백질의 발현 수준이 대조군 시료보다 높을 경우 삼중음성유방암의 전이 위험성이 높다고 판단하는 단계를 포함하는 삼중음성유방암 전이 예후 예측에 필요한 정보를 제공하는 방법.(3) determining that the risk of triple-negative breast cancer metastasis is high when the mRNA expression level of the CSDE1 gene or the CSDE1 protein expression level is higher than that of a control sample. A method for providing information necessary for predicting the prognosis of triple-negative breast cancer metastasis.
  12. (1) 삼중음성유방암 환자에서 분리된 시료로부터 CSDE1 유전자 및 RAC1 유전자의 mRNA 발현 수준 또는 CSDE1 단백질 및 RAC1 단백질의 발현 수준을 측정하는 단계; (1) measuring the mRNA expression level of CSDE1 gene and RAC1 gene or the expression level of CSDE1 protein and RAC1 protein from a sample isolated from a triple-negative breast cancer patient;
    (2) 상기 CSDE1 유전자 및 RAC1 유전자의 mRNA 발현 수준 또는 CSDE1 단백질 및 RAC1 단백질의 발현 수준을 대조군 시료와 비교하는 단계; 및(2) comparing the mRNA expression level of the CSDE1 gene and the RAC1 gene or the expression level of the CSDE1 protein and the RAC1 protein with a control sample; and
    (3) 상기 CSDE1 유전자 및 RAC1 유전자의 mRNA 발현 수준 또는 CSDE1 단백질 및 RAC1 단백질의 발현 수준이 대조군 시료보다 높을 경우 삼중음성유방암의 전이 위험성이 높다고 판단하는 단계를 포함하는 삼중음성유방암 전이 예후 예측에 필요한 정보를 제공하는 방법.(3) necessary for predicting the prognosis of triple-negative breast cancer metastasis, including the step of determining that the risk of metastasis of triple-negative breast cancer is high when the mRNA expression levels of the CSDE1 gene and the RAC1 gene or the expression levels of the CSDE1 protein and the RAC1 protein are higher than those of the control sample How to provide information.
  13. CSDE1 발현 또는 활성 억제제를 유효성분으로 포함하는 삼중음성유방암 예방 또는 치료용 약학조성물.A pharmaceutical composition for preventing or treating triple-negative breast cancer comprising a CSDE1 expression or activity inhibitor as an active ingredient.
  14. CSDE1 발현 또는 활성 억제제를 유효성분으로 포함하는 삼중음성유방암 전이 억제용 약학조성물.A pharmaceutical composition for inhibiting metastasis of triple-negative breast cancer comprising a CSDE1 expression or activity inhibitor as an active ingredient.
  15. 제14항에 있어서, 상기 약학조성물은 RAC1 발현을 억제하는 것을 특징으로 하는 삼중음성유방암 전이 억제용 약학조성물.The pharmaceutical composition for inhibiting triple-negative breast cancer metastasis according to claim 14, wherein the pharmaceutical composition inhibits RAC1 expression.
  16. (1) 삼중음성유방암 세포에 시험물질을 접촉시키는 단계;(1) contacting triple-negative breast cancer cells with a test substance;
    (2) 상기 시험물질을 접촉한 삼중음성유방암 세포에서 CSDE1의 발현 또는 활성 정도를 측정하는 단계; 및 (2) measuring the expression or activity level of CSDE1 in triple-negative breast cancer cells contacted with the test substance; and
    (3) 대조군 시료와 비교하여 상기 CSDE1의 발현 또는 활성 정도가 감소한 시험물질을 선별하는 단계를 포함하는 삼중음성유방암 치료제 스크리닝 방법.(3) A screening method for triple-negative breast cancer treatment comprising the step of selecting a test substance having a reduced expression or activity of the CSDE1 compared to a control sample.
  17. (1) 삼중음성유방암 세포에 시험물질을 접촉시키는 단계;(1) contacting triple-negative breast cancer cells with a test substance;
    (2) 상기 시험물질을 접촉한 삼중음성유방암 세포에서 CSDE1의 발현 또는 활성 정도를 측정하는 단계; 및 (2) measuring the expression or activity level of CSDE1 in triple-negative breast cancer cells contacted with the test substance; and
    (3) 대조군 시료와 비교하여 상기 CSDE1의 발현 또는 활성 정도가 감소한 시험물질을 선별하는 단계를 포함하는 삼중음성유방암 전이 억제제 스크리닝 방법.(3) A triple-negative breast cancer metastasis inhibitor screening method comprising the step of selecting a test substance having a reduced expression or activity level of the CSDE1 compared to a control sample.
  18. (1) 삼중음성유방암 세포에 시험물질을 접촉시키는 단계;(1) contacting triple-negative breast cancer cells with a test substance;
    (2) 상기 시험물질을 접촉한 삼중음성유방암 세포에서 CSDE1 및 RAC1의 발현 또는 활성 정도를 측정하는 단계; 및 (2) measuring the expression or activity levels of CSDE1 and RAC1 in triple-negative breast cancer cells contacted with the test substance; and
    (3) 대조군 시료와 비교하여 상기 CSDE1 및 RAC1의 발현 또는 활성 정도가 감소한 시험물질을 선별하는 단계를 포함하는 삼중음성유방암 전이 억제제 스크리닝 방법.(3) A triple-negative breast cancer metastasis inhibitor screening method comprising the step of selecting a test substance in which the expression or activity level of the CSDE1 and RAC1 is reduced compared to a control sample.
  19. CSDE1 발현 또는 활성 억제제를 유효성분으로 포함하는, 시험관(in vitro) 내 삼중음성유방암세포에서의 RAC1 발현 억제용 시약 조성물.A reagent composition for inhibiting RAC1 expression in triple-negative breast cancer cells in vitro, comprising a CSDE1 expression or activity inhibitor as an active ingredient.
  20. 시험관 내에서(in vitro) CSDE1 발현 또는 활성 억제제를 삼중음성유방암세포에 처리하는 단계를 포함하는 RAC1 발현 억제 방법.A method of inhibiting RAC1 expression comprising treating triple negative breast cancer cells with an inhibitor of CSDE1 expression or activity in vitro.
PCT/KR2022/009489 2021-07-12 2022-07-01 Biomarker composition for predicting triple negative breast cancer metastasis, containing csde1 as active ingredient WO2023287079A1 (en)

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Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20160291021A1 (en) * 2013-11-22 2016-10-06 Institut De Cancerologie De L'ouest Method for In Vitro Diagnosing and Prognosing of Triple Negative Breast Cancer Recurrence

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20160291021A1 (en) * 2013-11-22 2016-10-06 Institut De Cancerologie De L'ouest Method for In Vitro Diagnosing and Prognosing of Triple Negative Breast Cancer Recurrence

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
"Analysis of epigenetic modification change and discover the target for therapeutic agent development in triple-negative breast cancer", FINAL (RESULT) REPORT OF SUPPORT PROJECT FOR MID-SIZED RESEARCHERS, SOOKMYUNG WOMEN'S UNIVERSITY, KR, 1 January 2019 (2019-01-01), KR, pages 1 - 35, XP009543164 *
GUO AO-XIANG, CUI JIA-JIA, WANG LEI-YUN, YIN JI-YE: "The role of CSDE1 in translational reprogramming and human diseases", CELL COMMUNICATION AND SIGNALING, vol. 18, no. 1, 1 December 2020 (2020-12-01), XP093024141, DOI: 10.1186/s12964-019-0496-2 *
KIM YESOL, KO JE YEONG, LEE SOO-BEEN, OH SUMIN, PARK JEE WON, KANG HYEOK-GU, KIM DA-HYUN, CHUNG DAEUN, LIM SERA, KONG HYUNKYUNG, K: "Reduced miR-371b-5p expression drives tumor progression via CSDE1/RAC1 regulation in triple-negative breast cancer", ONCOGENE, NATURE PUBLISHING GROUP UK, LONDON, vol. 41, no. 22, 27 May 2022 (2022-05-27), London , pages 3151 - 3161, XP093024143, ISSN: 0950-9232, DOI: 10.1038/s41388-022-02326-6 *
WURTH LAURENCE; PAPASAIKAS PANAGIOTIS; OLMEDA DAVID; BLEY NADINE; CALVO GUADALUPE T.; GUERRERO SANTIAGO; CEREZO-WALLIS DANIEL: "UNR/CSDE1 Drives a Post-transcriptional Program to Promote Melanoma Invasion and Metastasis", CANCER CELL, CELL PRESS, US, vol. 30, no. 5, 27 October 2016 (2016-10-27), US , pages 694 - 707, XP029813184, ISSN: 1535-6108, DOI: 10.1016/j.ccell.2016.10.004 *

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