KR-102961344-B1 - Biomarker composition for diagnosing blood coagulation disorder comprising miRNA in extracellular vesicle

Abstract

The present invention relates to a biomarker for diagnosing blood coagulation disorders comprising miRNA within extracellular vesicles. It was experimentally confirmed that the miRNA within extracellular vesicles according to the present invention exhibits a specific expression pattern in patients with blood coagulation disorders compared to a normal group. Accordingly, this implies that the miRNA within extracellular vesicles of the present invention can be utilized as a biomarker for blood coagulation disorders and can be applied in various ways in the field of blood coagulation disorder diagnosis.

Inventors

• 방오영
• 김은희
• 성지희
• 신은경
• 정종원
• 서우근
• 김경문
• 안명주

Assignees

• (주)에스엔이바이오
• 사회복지법인 삼성생명공익재단
• 주식회사 제노헬릭스

Dates

Publication Date

20260511

Application Date

20230320

Priority Date

20220318

Claims (13)

1. A biomarker composition for diagnosing cancer-related stroke comprising one or more microvesicle-associated miRNAs selected from the group consisting of miR-205-5p, miR-645, and miR-646.
2. In paragraph 1, the cancer-related stroke diagnosis is i) Diagnosis of cancer-related stroke compared to the control group; ii) Diagnosis of cancer-related stroke compared to a group of stroke patients without cancer; and iii) a cancer-related stroke diagnosis; a biomarker composition comprising one or more diagnoses selected from a group consisting of cancer-related stroke diagnosis compared to a group of cancer patients without stroke.
3. A biomarker composition for diagnosing cancer-related stroke according to claim 1, wherein the composition comprises all of miR-205-5p, miR-645, and miR-646.
4. delete
5. A biomarker composition for diagnosing cancer-related stroke, wherein, in claim 1, the microvesicles are microvesicles in blood or serum.
6. delete
7. A biomarker composition for diagnosing cancer-related stroke according to claim 1, wherein miR-205-5p, miR-645, and miR-646 are highly expressed when cancer-related stroke occurs.
8. A composition for diagnosing cancer-related stroke comprising a preparation for measuring the expression level of one or more microvesicle-derived mRNAs selected from the group consisting of miR-205-5p, miR-645, and miR-646.
9. A composition for diagnosing cancer-related stroke according to claim 8, wherein the preparation is a primer or probe that specifically binds to one or more miRNAs selected from the group consisting of miR-205-5p, miR-645, and miR-646.
10. A cancer-related stroke diagnostic kit comprising the composition for diagnosing cancer-related stroke according to claim 8.
11. (a) a step of isolating microvesicles from a biological sample of the subject; and (b) a step of comparing the expression level of one or more miRNAs selected from the group consisting of miR-205-5p, miR-645, and miR-646 within the microvesicles with the expression level of the corresponding miRNA in a control sample; comprising A method for providing information for diagnosing cancer-related stroke, wherein step (b) determines that the subject has cancer-related stroke if the expression level of one or more miRNAs selected from the group consisting of miR-205-5p, miR-645, and miR-646 is high compared to the comparison control group.
12. In Clause 11, the above comparison control group is A method for providing information for the diagnosis of cancer-related stroke, comprising one or more types selected from a group consisting of a normal group, a group of stroke patients without cancer, and a group of cancer patients without stroke.
13. A method for providing information for the diagnosis of cancer-related stroke, wherein, in paragraph 11, the biological sample is blood or serum.

Description

Biomarker composition for diagnosing blood coagulation disorder comprising miRNA in extracellular vesicle The present invention relates to a biomarker for diagnosing blood coagulation disorders comprising miRNA within extracellular vesicles. Among the causes of death in patients with malignant tumors, in addition to death caused by the cancer itself, coagulation disorders resulting from various procoagulants secreted by cancer cells—such as cerebral infarction or pulmonary embolism—are the primary causes of death and disability. Various anticoagulants have been developed, and their use can effectively prevent and correct coagulation disorders. However, due to the lack of methods to predict these issues in cancer patients, the condition is currently diagnosed only after coagulation disorders have already occurred. Furthermore, since anticoagulants carry a risk of bleeding, it is difficult to recommend prophylactic administration in cancer patients who have not yet developed thrombosis. Therefore, while it is crucial to predict these issues in advance or diagnose them rapidly, no efficient preventive method has yet been identified to predict the risk of blood coagulation in cancer patients or to diagnose and prevent it early. Although various clinical predictive methods and blood D-dimer levels are utilized in clinical practice, they are non-specific and have low diagnostic value. Studies have been reported on diagnostic methods using blood microRNAs (miRNAs) to predict prognosis, including mortality, in patients with malignant tumors. However, since blood cell-free miRNAs are destroyed by blood RNases, a method is needed to measure more consistent miRNA levels. Extracellular vesicles are classified into exosomes and microvesicles based on their size; exosomes range in diameter from 30 to 150 nm, while microvesicles range from 100 to 1,000 nm. Extracellular vesicles are portions of cell membranes that have detached and entered the bloodstream. Containing both proteins and nuclear components, they are known to mediate intercellular communication. Since extracellular vesicles have been established to exist in most bodily fluids—such as blood, urine, saliva, breast milk, pleural fluid, ascites, and cerebrospinal fluid—either normally or during disease, their clinical utility is extremely high. Consequently, active international efforts are currently underway to develop diagnostic methods for various diseases, including cancer, by isolating and analyzing extracellular vesicles from bodily fluids. The miRNAs contained within extracellular vesicles are protected from circulating RNases by a lipid bilayer, resulting in consistently high levels of miRNAs compared to cell-free miRNAs. It is known that exosomes and microvesicles have different miRNA profiles regarding the miRNAs contained in extracellular vesicles. Figure 1 is a figure showing the overall study design. Figure 2A is a diagram showing the separation process of microvesicles and exosomes from extracellular vesicles in a patient's blood. Figure 2B shows the results of confirming the size distribution of extracellular vesicles through NanoSight Tracking Analysis. Figure 2C is a figure showing the results of confirming the morphology of extracellular vesicles using an electron microscope. Figure 2D shows the results of detecting extracellular vesicle positive and negative markers through Western blotting. Figure 3 shows the results of deriving candidate miRNAs associated with cancer-related coagulation disorders by measuring blood microvesicle and exosome miRNA levels in pooling samples from cancer patients with and without cerebral infarction (CC: cancer-control, CS: cancer-stroke). Figure 4a shows the results of comparing miRNA levels in microvesicles in samples from a normal person, cancer patients without cerebral infarction, and cancer patients with cerebral infarction. Figure 4b shows the ROC (Receiver operating characteristics) curve for cancer-stroke due to cancer-related coagulation disorders in cancer patients, based on the results of comparing miRNA levels within microvesicles in each sample. Figure 5A shows the results of comparing the levels of miR-205-5p, miR-645, and miR-646 in microvesicles of a group of stroke patients with cancer-related coagulation disorders (cancer-stroke) and a group of stroke patients without cancer (stroke-control, acute stroke). Figure 5B shows the ROC (Receiver operating characteristics) curve for cancer-stroke due to cancer-related coagulation disorders in cancer patients, based on the results of comparing the levels of miR-205-5p, miR-645, and miR-646 in microvesicles in each sample. Figure 6 shows the results of comparing mRNA expression using a Xeno-sensor technique in an independent cohort for external validation to diagnose cerebral infarction due to cancer-related coagulation disorders (A, B) and the results of diagnosing cerebral infarction due to cancer-related coagulation disorders (C). Figure 7A shows the