WO2022025625A1 - Composition de biomarqueur destinée à une détection de cellules souches cancéreuses - Google Patents

Composition de biomarqueur destinée à une détection de cellules souches cancéreuses Download PDF

Info

Publication number
WO2022025625A1
WO2022025625A1 PCT/KR2021/009800 KR2021009800W WO2022025625A1 WO 2022025625 A1 WO2022025625 A1 WO 2022025625A1 KR 2021009800 W KR2021009800 W KR 2021009800W WO 2022025625 A1 WO2022025625 A1 WO 2022025625A1
Authority
WO
WIPO (PCT)
Prior art keywords
cancer
cells
cancer stem
phosphatidylethanolamine
stem cells
Prior art date
Application number
PCT/KR2021/009800
Other languages
English (en)
Korean (ko)
Inventor
김재호
신민주
김대경
Original Assignee
부산대학교 산학협력단
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 부산대학교 산학협력단 filed Critical 부산대학교 산학협력단
Publication of WO2022025625A1 publication Critical patent/WO2022025625A1/fr

Links

Images

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
    • G01N33/57492—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites involving compounds localized on the membrane of tumor or cancer cells
    • 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
    • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • 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
    • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56966—Animal cells
    • 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

Definitions

  • the present invention relates to a biomarker for detecting cancer stem cells, and more particularly, a biomarker composition for detecting cancer stem cells using phosphatidylethanolamine (PE) expressed in cancer stem cells, a composition for cancer diagnosis, a cancer diagnosis kit and to a method of providing information on cancer diagnosis.
  • PE phosphatidylethanolamine
  • stem cells are essential for maintaining and regenerating the organs of the body.
  • Tumors also contain cancer stem cells, a heterogeneous population with stem cell-like properties.
  • the cancer stem cells are cells that form spheres using the self-renewal method, which is a characteristic of cancer stem cells, through floating culture, and are known to share strong cancer-forming ability and characteristics of normal stem cells compared to the original cell line.
  • cancer stem cells tend to increase as the tumor progresses to malignancy, and cancer stem cells are reported to cause cancer as well as resistance to existing cancer therapies and cause recurrence. Therefore, in order to remove cancer stem cells, it is important to recognize and target cells resistant to existing anticancer drugs.
  • cancer stem cells are known as the highest-level cells exhibiting these heterogeneous characteristics, and are closely related to anticancer drug resistance. Therefore, it is not easy to completely remove cancer stem cells despite various cell surface protein-targeted treatments. Most of the current cancer stem cell protein target biomarkers can have different expression frequencies depending on various surrounding environments, such as cell signal transduction or cell resistance acquisition. Therefore, it is important to find a cancer stem cell-specific cell surface target biomarker other than the existing protein target biomarker.
  • the present inventors completed the present invention by confirming that the expression of phosphatidylethanolamine in ovarian cancer cancer stem cells was significantly higher than that of ovarian cancer cell lines and other cell lines as a result of research to discover biomarkers for targeting cancer stem cells. did
  • an object of the present invention is to provide a biomarker composition for detecting cancer stem cells containing phosphatidylethanolamine (PE).
  • PE phosphatidylethanolamine
  • Another object of the present invention is to provide a composition for diagnosing cancer through detection of cancer stem cells, including an agent for measuring the expression of phosphaditylethanolamine, and a cancer diagnostic kit comprising the same.
  • Another object of the present invention is to provide a method for providing information on cancer diagnosis through cancer stem cell detection, comprising measuring the expression of phosphatidylethanolamine in a biological sample.
  • the present invention provides a biomarker composition for detecting cancer stem cells comprising phosphatidylethanolamine.
  • the present invention provides a composition for diagnosing cancer through cancer stem cell detection comprising an agent for measuring the expression of phosphatidylethanolamine.
  • the present invention also provides a cancer diagnosis kit through cancer stem cell detection comprising the composition for cancer diagnosis.
  • the present invention also provides a method for providing information on cancer diagnosis through cancer stem cell detection, comprising measuring the expression of phosphatidylethanolamine in a biological sample.
  • phosphatidylethanolamine a biomarker for cancer stem cell detection according to the present invention
  • FIG. 1 is a diagram showing the results of morphological analysis of ovarian cancer cell line A2780-AD cells and ovarian cancer stem cell line A2780-SP cells.
  • FIG. 2 is a diagram showing the results of confirming the expression of cancer stem cell markers in ovarian cancer cell line A2780-AD cells and ovarian cancer stem cell line A2780-SP cells through flow cytometry.
  • FIG. 3 is a diagram showing the results of analyzing the lipid expression of ovarian cancer cell line A2780-AD cells and ovarian cancer stem cell line A2780-SP cells through LC-MS.
  • FIG. 4 is a diagram showing the results of analysis of lipids with increased or decreased expression in A2780-SP cells, a cancer stem cell line, compared to A2780-AD, an ovarian cancer cell line, based on the results of FIG. 3 .
  • FIG. 5 is a diagram showing the results of comparing the expression of phosphatidylethanolamine in ovarian cancer cell line A2780-AD cells and ovarian cancer stem cell line A2780-SP cells.
  • FIG. 6 is a diagram showing the results of analyzing the lipid expression of cancer stem cell line A2780-DIF cells and cancer stem cell line A2780-SP cells that have been cancerousized through LC-MS.
  • A2780-DIF cells which are cancer stem cell lines, which have become cancerous compared to A2780-SP cells, which are cancer stem cell lines.
  • FIG. 8 is a diagram showing the results of comparing the expression of phosphatidylethanolamine in cancer stem cell line A2780-DIF cells and cancer stem cell line A2780-SP cells transformed into cancer cells.
  • EOC-SP cells which are patient-derived ovarian cancer stem cells
  • EOC-DIF cells which are patient-derived ovarian cancer stem cells, which have become cancerous.
  • FIG. 10 is a diagram showing the results of confirming the expression of cancer stem cell markers in EOC-SP cells and EOC-DIF cells.
  • FIG. 11 is a diagram showing the results of analyzing the lipid expression of EOC-SP cells and EOC-DIF cells.
  • EOC-DIF cells which are cancer stem cell lines, which have become cancerous compared to EOC-SP cells, which are patient-derived ovarian cancer cancer stem cell lines, based on the results of FIG. 11 . is the diagram shown.
  • FIG. 13 is a diagram showing the results of comparing the expression of phosphatidylethanolamine in EOC-SP cells and EOC-DIF cells.
  • FIG. 14 is a diagram showing the results of confirming whether phosphatidylethanolamine is exposed to the outside of the cell membrane in ovarian cancer cell line A2780-AD cells and ovarian cancer stem cell line A2780-SP cells through flow cytometry.
  • FIG. 15 is a diagram showing the results of confirming whether phosphatidylethanolamine is exposed to the outside of the cell membrane in ovarian cancer cell line SKOV3-AD cells and ovarian cancer stem cell line SKOV3-SP cells through flow cytometry.
  • 16 is a diagram showing the results of confirming whether phosphatidylethanolamine is exposed to the outside of the cell membrane in the patient-derived cancer stem cell lines EOC12-SP cells and EOC21-SP cells through flow cytometry.
  • 17 is a diagram showing the results of confirming the expression of phosphatidylethanolamine and external exposure to the cell membrane in human embryonic kidney cells, tonsil-derived mesenchymal stem cells, and human umbilical vein endothelial cells through flow cytometry.
  • a biomarker composition for detecting cancer stem cells comprising phosphatidylethanolamine (PE).
  • PE phosphatidylethanolamine
  • phosphatidylethanolamine is one of phospholipids, and belongs to glycerophospholipids.
  • the glycerophospholipid is a form in which two fatty acids are bonded to glycerol 3-phosphate, and phosphatidylethanolamine is a phosphoric acid group in which ethanolamine is bonded.
  • Such phosphoethanolamine is known to be involved in membrane fusion during cell division, decomposition of contractile rings, and control of membrane curvature.
  • the phosphatidylethanolamine is generally expressed in the inner membrane of the phospholipid bilayer of the cell.
  • cancer stem cell (Cancer stem cell or Tumor initiating cell) refers to a cell having the ability to generate a tumor. Cancer stem cells have characteristics similar to normal stem cells, and they generate tumors through the self-renewal and differentiation abilities of stem cells. In addition, the tumor is differentiated from other populations and causes recurrence and metastasis by generating new tumors. Therefore, the development of a specific treatment method targeting cancer stem cells can increase the survival rate of cancer patients.
  • cancer is defined as an aggressive characteristic in which cells divide and grow ignoring normal growth limits, an invasive characteristic that penetrates into surrounding tissues, and metastatic that spreads to other parts of the body ) is a generic term for diseases caused by cells with characteristics.
  • gastric cancer gastric cancer, breast cancer, lung cancer, liver cancer, blood cancer, bone cancer, pancreatic cancer, Skin cancer, head or neck cancer, cutaneous or intraocular melanoma, uterine sarcoma, ovarian cancer, rectal cancer, anal cancer (anal cancer), colon cancer, fallopian tube carcinoma, endometrial carcinoma, cervical cancer, small intestine cancer, endocrine cancer, Thyroid cancer, parathyroid cancer, kidney cancer, soft tissue tumor, urethral cancer, prostate cancer, bronchogenic cancer and bone marrow cancer It is preferably at least one selected from the group consisting of (bone marrow tumor), and more preferably ovarian cancer.
  • a “biomarker” is an index that can objectively measure and evaluate changes in a living body, and can measure and evaluate an individual's pathological state, response to drugs, and the like. Accordingly, the biomarker may also be used for detection or diagnosis.
  • the phosphatidylethanolamine is preferably expressed in non-cancer cells and cancer stem cells than in cancer cells.
  • non-cancer cells may include, but are not limited to, human embryonic-derived kidney cells, tonsil-derived mesenchymal stem cells, and human umbilical vein endothelial cells.
  • phosphatidylethanolamine is expressed on the surface of cancer stem cells and is overexpressed in cancer stem cells compared to non-cancer cells and cancer cells, which is a cancer stem cell detection or cancer diagnosis bio It can be used as a marker.
  • composition for diagnosing cancer through detection of cancer stem cells and a cancer diagnosis kit comprising an agent for measuring the expression of phosphatidylethanolamine.
  • diagnosis means confirming the presence or characteristics of a pathological condition.
  • the diagnosis is meant to include not only whether or not cancer occurs, but also confirmation of prognosis, progress, and stage.
  • diagnosis means identifying the onset, prognosis, course, staging or characteristics of cancer.
  • the “agent for measuring the expression of phosphatidylethanolamine” specifically binds to phosphatidylethanolamine, a biomarker whose expression is increased in cancer stem cells, as described above, and confirms the expression level of the biomarker. It means that it can be used for detection.
  • the agent for measuring the expression of phosphatidylethanolamine is an oligopeptide that specifically binds to phosphatidylethanolamine, a monoclonal antibody, a polyclonal antibody, a chimeric antibody, a ligand, PNA (Peptide nucleic acid), an aptamer (aptamer), preferably selected from the group consisting of probes and dyes, but is not limited thereto.
  • the agent for measuring the expression of phosphatidylethanolamine may include duramycin or cinnamycin.
  • the kit for diagnosis of cancer may be a kit in various types depending on the method used, for example, the kit is quantitative analysis for flow cytometry, mass spectrometry, light absorption spectrometry, chromatography and emission spectroscopy. It may be a kit, but is not limited thereto.
  • the quantitative analysis kit may obtain a result value through a quantitative analysis device.
  • the quantitative analysis device may be at least one selected from the group consisting of flow cytometry, nuclear magnetic resonance spectroscopy, chromatography, ultraviolet spectroscopy, infrared spectroscopy, fluorescence spectroscopy, microplate reader and mass spectrometer, but is not limited thereto.
  • the cancer diagnostic kit may include not only an agent for measuring the expression level of the phosphatidylethanolamine, but also tools and reagents commonly used in the art used for immunological analysis and quantitative analysis.
  • the tool or reagent include, but are not limited to, a suitable carrier, a labeling material capable of generating a detectable signal, chromophores, solubilizers, detergents, buffers, stabilizers, and the like.
  • the labeling material is an enzyme, it may include a substrate capable of measuring enzyme activity and a reaction terminator.
  • the carrier includes a soluble carrier and an insoluble carrier
  • an example of the soluble carrier is a physiologically acceptable buffer known in the art, for example, PBS
  • examples of the insoluble carrier include polystyrene, polyethylene, polypropylene, polyester, poly It may be acrylonitrile, fluororesin, crosslinked dextran, polysaccharide, polymer such as magnetic fine particles plated with metal in latex, other paper, glass, metal, agarose, and combinations thereof.
  • kit of the present invention includes the above-described composition as a component, redundant descriptions are omitted in order to avoid excessive complexity of the present specification.
  • cancer stem cells that is, cells overexpressing phosphatidylethanolamine
  • a biological sample to determine the presence of cancer stem cells, and from this, the presence of cancer as well as However, the prognosis, course, and stage can be confirmed.
  • the present invention provides a method for providing information on cancer diagnosis through cancer stem cell detection, comprising measuring the expression of phosphatidylethanolamine in a biological sample.
  • biological sample means any sample obtained from an individual in which the expression of phosphatidylethanolamine of the present invention can be detected.
  • the biological sample is any one selected from the group consisting of whole blood, serum, plasma, cells, sputum, tissue, saliva, biopsy, liquid culture, feces and urine, and is not particularly limited thereto, and the present invention It can be prepared by processing by a method commonly used in the technical field of
  • the method determines the presence or absence of cancer stem cells by checking the expression level of phosphatidylethanolamine in a biological sample, thereby providing information on diagnosis of cancer.
  • the method comprises at least one quantitative analysis device selected from the group consisting of flow cytometry, nuclear magnetic resonance spectroscopy, chromatography, ultraviolet spectroscopy, infrared spectroscopy, fluorescence spectroscopy, microplate reader and mass spectrometer. It may further comprise the step of confirming the expression of tylethanolamine.
  • the present invention comprises the steps of (a) obtaining an assay sample from a subject; (b) detecting the presence or absence of phosphatidylethanolamine (PE) in the analysis sample of step (a); (c) diagnosing cancer when phosphatidylethanolamine is present in the analyte sample in step (b); (d) administering an effective amount of a cancer stem cell-specific anticancer agent to the individual diagnosed with cancer in step (c); provides a cancer diagnosis or treatment method comprising a.
  • PE phosphatidylethanolamine
  • the cancer stem cell-specific anticancer agent is nitrogen mustard, imatinib, oxaliplatin, rituximab, erlotinib, neratinib, lapatinib, gefitinib, vandetanib, nirotinib, semasanib , bosutinib, axitinib, cediranib, restaurtinib, trastuzumab, gefitinib, bortezomib, sunitinib, carboplatin, sorafenib, bevacizumab, cisplatin, cetuximab, viscumalbum , asparaginase, tretinoin, hydroxycarbamide, dasatinib, estramustine, gemtuzumab ozogamicin, ibritumomab tusetan, heptaplatin, methylaminolevulinic acid, ams
  • Example 1 Comparison of lipid expression of ovarian cancer cells and ovarian cancer stem cells
  • Ovarian cancer cell line A2780 cells (A2780-AD) and ovarian cancer stem cell line A2780-SP cells were prepared.
  • the ovarian cancer cell line A2780-AD was cultured at 37° C. and 5% CO 2 using RPMI1640 medium (10% FBS, 100 ⁇ g/mL Sterptomycin/Penicillin), and subcultured at regular intervals.
  • the cancer stem cell line A2780-SP cells are obtained by three-dimensional culture of the ovarian cancer cell line A2780 cells, followed by isolation of cancer stem cells.
  • the three-dimensional culture was performed in an ultra-low attachent dish using Neurobasal Plus medium (B-27 supplement (50X), 100 ⁇ g/mL Sterptomycin/Penicillin, 100 ⁇ g/mL rhFGF, 100 ⁇ g/mL rhEGF, Glutamax, 1M HEPES). was performed in The three-dimensional culture conditions are a temperature of 37° C. and 5% CO 2 conditions.
  • Morphological analysis of the prepared A2780-AD cells and A2780-SP cells was performed. Specifically, the A2780-AD cells and A2780-SP cells were cultured under each of the above conditions. The cultured cells were observed under a microscope. The morphological analysis results are shown in FIG. 1 .
  • aldehyde dehydrogenase (ALDH), a marker of cancer stem cells, was confirmed in the prepared A2780-AD cells and A2780-SP cells. Specifically, using the ALDEFOUORTM kit (STEMCELL Technologies), ALDH-expressing cells among the prepared A2780-AD cells were labeled according to the manufacturer's manual. Thereafter, cells expressing ALDH were measured using FACS AttuneNxT (ThermoFisher). A2780-SP cells were also measured for ALDH expression in the same manner. The results of measuring cells expressing cancer stem cell markers are shown in FIG. 2 .
  • A2780-SP cells which is an ovarian cancer stem cell line
  • A2780-AD ovarian cancer cell line
  • A2780-AD cells Lipid expression of the prepared A2780-AD cells and A2780-SP cells was analyzed. Specifically, adherent A2780-AD cells were treated with trypsin to collect cell pellets. In addition, A2780-SP cells, which are floating cells, were spun down after recovering the culture medium to remove the medium to collect cell pellets. MeOH 610 ⁇ l, internal standards mix 50 ⁇ l, chloroform 330 ⁇ l and Cholesteryl ester standard (1 ng/ ⁇ l) 20 ⁇ l were added to the collected cell pellet. After that, after vortexing for 30 seconds every 3 minutes, spin down and sonication were performed, and repeated 3 times. The sonicated cells were centrifuged (4°C, 1 min, 14,000 g), and the supernatant was removed to obtain a pellet.
  • sample contained in a new tube was divided into samples for non-methylation and methylation analysis, and then dried.
  • TMSD Trimethylsilyldiazomethane
  • Lipids contained in the prepared sample were analyzed through liquid chromatography mass spectrometry (LC-MS).
  • LC-MS liquid chromatography mass spectrometry
  • lipids with increased or decreased expression in A2780-SP cells were analyzed compared to A2780-AD, an ovarian cancer cell line.
  • SigmaPlot10.0 software the LC-MS raw data was plotted to analyze lipids with increased or decreased expression. The results of lipid expression analysis are shown in FIG. 4 .
  • lipids with increased expression in the ovarian cancer stem cell line A2780-SP cells compared to the ovarian cancer cell line A2780-AD are phosphatidylethanolamine (PE), lysophosphatidylethanolamine (LPE), mono It was confirmed that there are 93 types including glyceride (monoglyceride), and the lipids with reduced expression are 61 types including phosphatidylcholine, lysophosphatidylcholine, and diglyceride.
  • PE phosphatidylethanolamine
  • LPE lysophosphatidylethanolamine
  • the ovarian cancer stem cell line A2780-SP cells were cultured under cancer cell culture conditions to form cancer cells.
  • the cancerous ovarian cancer stem cells were named A2780-DIF.
  • Lipid expression of the A2780-DIF cells and ovarian cancer stem cells (A2780-SP) was confirmed in the same manner as in Examples 1-4.
  • a matrix was prepared based on the analysis results, and the created matrix is shown in FIG. 6 .
  • A2780-SP cells and A2780-DIF cells had different lipid expression patterns.
  • the lipid expression pattern of A2780-DIF cells was somewhat different from that of the ovarian cancer cell line A2780-AD, but had a similar tendency.
  • lipids with increased or decreased expression in A2780-DIF compared to A2780-SP cells, a cancer stem cell line, were analyzed, and the results are shown in FIG. 7 .
  • lipids with reduced expression in the ovarian cancer cell line A2780-DIF compared to the ovarian cancer stem cell line A2780-SP cells are 87 types including phosphatidylethanolamine (PE), and the lipids with increased expression are It was confirmed that phosphatidylcholine (phosphatidylcholine), lysophosphatidylcholine (lysophosphatidylcholine), diglyceride (diglyceride), such as 84 kinds.
  • PE phosphatidylethanolamine
  • Example 2 Comparison of lipid expression of patient-derived ovarian cancer cancer stem cells and cancerousized patient-derived ovarian cancer cancer stem cells
  • Ovarian cancer stem cells were isolated from the patient and named as EOC-SP.
  • the EOC-SP cells were cultured under cancer cell culture conditions to form cancer cells.
  • the ovarian cancer stem cells derived from the cancerousized patient were named EOC-DIF.
  • the prepared EOC-SP cells and EOC-DIF cells were used in the experiments described below.
  • EOC-SP cells and EOC-DIF cells were prepared EOC-SP cells and EOC-DIF cells. Specifically, the cells were each three-dimensionally cultured and then observed under a microscope. The morphological analysis results are shown in FIG. 9 .
  • EOC-SP cells and EOC-DIF cells were morphologically different.
  • the EOC-DIF cells were cancer cells, and showed a morphology similar to that of cancer cells.
  • aldehyde dehydrogenase a marker of cancer stem cells
  • EOC-SP cells and EOC-DIF cells were confirmed in EOC-SP cells and EOC-DIF cells. Specifically, using the ALDEFOUORTM kit (STEMCELL Technologies), ALDH-expressing cells among EOC-SP cells were labeled according to the manufacturer's manual. Thereafter, cells expressing ALDH were measured using FACS AttuneNxT (ThermoFisher). EOC-DIF cells were also measured for ALDH expression in the same manner. The results of measuring cells expressing cancer stem cell markers are shown in FIG. 10 .
  • EOC-SP cells and EOC-DIF cells Lipid expression of EOC-SP cells and EOC-DIF cells was analyzed. Specifically, adherent EOC-DIF cells were treated with trypsin to collect cell pellets. EOC-SP cells, which are floating cells, were spun down after recovering the culture medium to remove the medium to collect cell pellets. The collected cells were analyzed for lipid expression in the same manner as in Examples 1-4. A matrix was prepared based on the analysis results, and the created matrix is shown in FIG. 11 .
  • EOC-SP cells and EOC-DIF cells had different lipid expression patterns.
  • Lipids with increased or decreased expression in EOC-DIF cells a patient-derived ovarian cancer stem cell line that have become cancerous compared to EOC-SP cells, were analyzed, and the results are shown in FIG. 12 .
  • lipids with increased expression in EOC-SP cells compared to EOC-DIF cells are 87 types, including phosphatidylethanolamine, phosphatidylcholine, and lysophosphatidylcholine, and lipids with reduced expression are It was confirmed that 84 types including phosphatidylserine.
  • EOC-SP cells which are patient-derived ovarian cancer stem cells
  • Example 3 Analysis of exposure to phosphatidylethanolamine in ovarian cancer cancer stem cells
  • A2780-AD ovarian cancer cell line
  • A2780-SP ovarian cancer stem cell line
  • Example 3-1 Through flow cytometry, the expression of phosphatidylethanolamine in SKOV3-AD cells, another ovarian cancer cell line, and SKOV3-SP cells, a cancer stem cell line thereof, was analyzed in the same manner as in Example 3-1.
  • the ovarian cancer stem cell line SKOV3-SP cells are isolated after three-dimensional culture of the ovarian cancer cell line SKOV3-AD cells.
  • the flow cytometry results are shown in FIG. 15 .
  • the expression of phosphatidylethanolamine in the ovarian cancer stem cell line SKOV3-SP cells was significantly higher than that of the ovarian cancer cell line SKOV3-AD, and it was confirmed that the expressed phosphatidylethanolamine was exposed to the outside of the cell membrane.
  • EOC12-SP cells and EOC21-SP cells were prepared.
  • the expression of phosphatidylethanolamine in the EOC12-SP cells and the EOC21-SP cells was analyzed in the same manner as in Example 3-1.
  • the flow cytometry results are shown in FIG. 16 .
  • the patient-derived ovarian cancer stem cell lines, EOC12-SP cells and EOC21-SP cells both had high levels of phosphatidylethanolamine expression, and it was confirmed that the expressed phosphatidylethanolamine was exposed to the outside of the cell membrane.
  • TMSC, HEK293FT and HUVEC showed very little expression of phosphatidylethanolamine, and it was confirmed that the expressed phosphatidylethanolamine was exposed to the outside of the cell membrane.
  • phosphatidylethanolamine can be used as a biomarker for cancer stem cell detection, and the phosphatidylethanol of the present invention can be variously used in the fields of cancer diagnosis, prognosis prediction, and drug screening.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Urology & Nephrology (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Chemical & Material Sciences (AREA)
  • Hematology (AREA)
  • Cell Biology (AREA)
  • Medicinal Chemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Food Science & Technology (AREA)
  • Pathology (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Oncology (AREA)
  • Virology (AREA)
  • Hospice & Palliative Care (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Zoology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

La présente invention concerne un biomarqueur permettant de détecter des cellules souches cancéreuses. Plus particulièrement, la présente invention concerne : une composition de biomarqueur permettant de détecter des cellules souches cancéreuses à l'aide de la phosphatidyléthanolamine (PE) exprimée dans les cellules souches cancéreuses ; une composition de diagnostic du cancer ; un kit de diagnostic du cancer ; et une méthode permettant de fournir des informations sur le diagnostic du cancer. Il a été découvert que la phosphatidyléthanolamine, un biomarqueur permettant la détection de cellules souches cancéreuses selon la présente invention, est exprimée à des niveaux significativement plus élevés dans des cellules souches cancéreuses que dans des cellules cancéreuses et d'autres cellules, et est exposée à l'extérieur de la membrane cellulaire lorsqu'elle est exprimée. Cela signifie que des cellules souches cancéreuses peuvent être détectées avec une sensibilité et une précision plus élevées lorsque le biomarqueur permettant la détection de cellules souches cancéreuses selon la présente invention est utilisé, et ainsi la présente invention peut être utilisée de diverses manières dans les domaines du diagnostic, de la prédiction de pronostic du cancer et du criblage de médicaments contre le cancer.
PCT/KR2021/009800 2020-07-28 2021-07-28 Composition de biomarqueur destinée à une détection de cellules souches cancéreuses WO2022025625A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR10-2020-0093533 2020-07-28
KR1020200093533A KR102400464B1 (ko) 2020-07-28 2020-07-28 암 줄기세포 검출용 바이오 마커 조성물

Publications (1)

Publication Number Publication Date
WO2022025625A1 true WO2022025625A1 (fr) 2022-02-03

Family

ID=80035852

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2021/009800 WO2022025625A1 (fr) 2020-07-28 2021-07-28 Composition de biomarqueur destinée à une détection de cellules souches cancéreuses

Country Status (2)

Country Link
KR (1) KR102400464B1 (fr)
WO (1) WO2022025625A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114814023A (zh) * 2022-04-24 2022-07-29 江苏省中医院 脂质分子在作为胃癌化疗药耐药的预测性标志物中的应用

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20150035821A (ko) * 2012-06-15 2015-04-07 해리 스타일리 질환 또는 병태를 검출하는 방법
JP5702366B2 (ja) * 2009-05-11 2015-04-15 セレクター，インコーポレイティド 蛍光リン脂質エーテル化合物、組成物、及びその使用
KR20170041192A (ko) * 2014-06-25 2017-04-14 텔 하쇼머 메디컬 리서치 인프라스트럭쳐 앤드 서비시스 리미티드． 암 줄기세포 마커의 동정 및 진단 및 치료를 위한 그의 사용
KR20180010938A (ko) * 2016-07-22 2018-01-31 성균관대학교산학협력단 영상화용 발광 입자체, 이를 이용한 세포 영상화 방법, 및 이의 제조방법
KR20190037226A (ko) * 2016-06-14 2019-04-05 셀렉타 바이오사이언시스, 인코퍼레이티드 순환 종양 세포의 식별 및 단리를 위한 인지질 에테르 유사체

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2281578A3 (fr) 2002-07-15 2011-04-13 Board of Regents, The University of Texas System Anticorps et peptides de duramycine sélectionnés se fixant à des phospholipides et des aminophospholipides anioniques et leur utilisation dans le traitement d'infections virales et du cancer

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP5702366B2 (ja) * 2009-05-11 2015-04-15 セレクター，インコーポレイティド 蛍光リン脂質エーテル化合物、組成物、及びその使用
KR20150035821A (ko) * 2012-06-15 2015-04-07 해리 스타일리 질환 또는 병태를 검출하는 방법
KR20170041192A (ko) * 2014-06-25 2017-04-14 텔 하쇼머 메디컬 리서치 인프라스트럭쳐 앤드 서비시스 리미티드． 암 줄기세포 마커의 동정 및 진단 및 치료를 위한 그의 사용
KR20190037226A (ko) * 2016-06-14 2019-04-05 셀렉타 바이오사이언시스, 인코퍼레이티드 순환 종양 세포의 식별 및 단리를 위한 인지질 에테르 유사체
KR20180010938A (ko) * 2016-07-22 2018-01-31 성균관대학교산학협력단 영상화용 발광 입자체, 이를 이용한 세포 영상화 방법, 및 이의 제조방법

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114814023A (zh) * 2022-04-24 2022-07-29 江苏省中医院 脂质分子在作为胃癌化疗药耐药的预测性标志物中的应用

Also Published As

Publication number Publication date
KR102400464B1 (ko) 2022-05-19
KR20220014003A (ko) 2022-02-04

Similar Documents

Publication Publication Date Title
US11959838B2 (en) Representative diagnostics
WO2017020617A1 (fr) Procédé et dispositif de détection de cellule tumorale circulante
US11768137B2 (en) Size-based separation of dissociated fixed tissues
Berardocco et al. RNA-seq reveals distinctive RNA profiles of small extracellular vesicles from different human liver cancer cell lines
US20130288248A1 (en) Cancer stem cell mass and process for production thereof
US20070196345A1 (en) Antibody recognizing proliferative human liver cells, proliferative human liver cells and functional human liver cells
CN111073846B (zh) 一种分离组织特异来源细胞外囊泡的方法及其试剂盒
WO2022025625A1 (fr) Composition de biomarqueur destinée à une détection de cellules souches cancéreuses
WO2020206891A1 (fr) Hexokinase 2 utilisée à des fins de détection et kit de test pour des cellules néoplasiques rares dans un échantillon de liquide corporel
CN106771185A (zh) 一种鼻咽癌循环肿瘤细胞检测试剂盒
Esparza-López et al. Deriving primary cancer cell cultures for personalized therapy
Lu et al. Evaluation of a new magnetic bead as an integrated platform for systematic CTC recognition, capture and clinical analysis
Zhang et al. Clinical significance of Fusobacterium nucleatum infection and regulatory T cell enrichment in esophageal squamous cell carcinoma
Dang et al. Cancer-associated fibroblasts are key determinants of cancer cell invasion in the earliest stage of colorectal cancer
WO2019035565A2 (fr) Procédé de dépistage d'un patient atteint d'un cancer de la prostate basé sur un variant de récepteur d'androgènes
CN111344570A (zh) 分离和检测癌症干细胞的方法
CN113939584A (zh) 外泌体液体活检样本的制备装置和制备方法以及由此制备的外泌体液体活检样本的分析方法
Wei et al. Epithelial-mesenchymal transition classification of circulating tumor cells predicts clinical outcomes in progressive nasopharyngeal carcinoma
WO2022102885A1 (fr) Technique de diagnostic du stress associé à la dermatite atopique utilisant des miarn dérivés d'exosomes
Hu et al. Evaluation of Circulating Tumor Cells-DNA Methylated Separation Nano-Polylactic-co-Glycolic Acid Magnetic Beads with Epithelial Cell Adhesion Molecul/Epidermal Growth Factor Receptor Double Antibody Modification and Its Application in Lung Cancer Diagnosis
US20190317075A1 (en) INTERTUMORAL HOMOGENEITY DETERMINED BY MiCK ASSAY
WO2019035623A2 (fr) Méthode de diagnostic d'un cancer au moyen de nanoparticules liées à la lectine
WO2021194145A1 (fr) Composition pour l'extraction d'adn acellulaire comprenant un complexe alginate-polydopamine-silice
Zhou et al. Liposome–exosome hybrids for in situ detection of exosomal miR-1246 in breast cancer
US20240085306A1 (en) Method for enriching cells or cell nuclei

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 21848691

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 21848691

Country of ref document: EP

Kind code of ref document: A1