Cancer-derived microvesicle-associated microrna as a diagnostic marker
View Patent ↗The presently disclosed subject matter provides methods of diagnosis of cancer or adverse pregnancy outcomes in a subject by measuring amounts of one or more microRNAs present in cancer-derived exosomes isolated from a biological sample from the subject.
1. A method for assessing the presence of one or more microRNAs in microvesicles, comprising isolating a population of cancer-derived microvesicles from a biological sample using a microvesicle surface marker, isolating microRNA from said population of cancer-derived microvesicles and determining a presence of one or more microRNAs in said cancer-derived microvesicles.
2. A method of determining the presence of one or more bio-markers in microvesicles, comprising: isolating a population of cancer-derived extracellular microvesicles from a biological sample; isolating microRNA from said population of cancer-derived extracellular microvesicles and determining an expression profile of one or more microRNA; and comparing the expression profile with a profile from a selected reference sample to determine a presence of one or more biomarkers in the microvesicles.
3. The method of claim 1 , wherein the biological sample is from a human.
4. The method of claim 1 , wherein the biological sample comprises milk, blood, serum, plasma, ascites, cyst fluid, pleural fluid, peritoneal fluid, cerebral spinal fluid, tears, urine, saliva, sputum, or combinations thereof.
5. The method of claim 1 , wherein the surface marker is EpCAM.
6. The method of claim 1 , wherein the surface marker is selected from a group consisting of EpCAM, Fas ligand, PD-1, MICA/B, mdr 1, MMPs, CD44, autoreactive antigens, tetraspanins, and MHC class I molecule.
7. The method of claim 1 , wherein said determining step comprises labeling the one or more microRNAs with a detectable label.
8. The method of claim 1 , wherein said determining comprises capturing the one or more microRNAs with one or more polynucleotide probes that selectively bind each of the one or more microRNAs.
9. The method of claim 1 , wherein said determining comprises using a real-time polymerase chain reaction.
10. The method of claim 1 , wherein the one or more microRNAs comprise one or more microRNAs set forth in Table 1.
11. The method of claim 1 , wherein the one or more microRNAs comprise one or more microRNAs set forth in Table 2.
12. The method of claim 1 , wherein the isolating comprises using size exclusion chromatography, filtration or immunosorbent capture.
13. The method of claim 1 , wherein the microRNA comprises one or more microRNAs selected from the group consisting of miR-21, miR-141, miR-200a, miR-200b, miR-200c, miR-203, miR-205, and miR-214.
14. The method of claim 2 , wherein the biological sample is from a human.
15. The method of claim 2 , wherein the biological sample comprises milk, blood, serum, plasma, ascites, cyst fluid, pleural fluid, peritoneal fluid, cerebral spinal fluid, tears, urine, saliva, sputum, or combinations thereof.
16. The method of claim 2 , wherein said determining step comprises labeling the one or more microRNAs with a detectable label.
17. The method of claim 2 , wherein said determining comprises capturing the one or more microRNAs with one or more polynucleotide probes that selectively bind each of the one or more microRNAs.
18. The method of claim 2 , wherein said determining comprises using a real-time polymerase chain reaction.
19. The method of claim 2 , wherein the one or more microRNAs comprise one or more microRNAs set forth in Table 1.
20. The method of claim 2 , wherein the one or more microRNAs comprise one or more microRNAs set forth in Table 2.
21. The method of claim 2 , wherein the isolating comprises using size exclusion chromatography, filtration or immunosorbent capture.
22. The method of claim 2 , wherein the microRNA comprises one or more microRNAs selected from the group consisting of miR-21, miR-141, miR-200a, miR-200b, miR-200c, miR-203, miR-205, and miR-214.
23. The method of claim 9 , wherein the surface marker is EpCAM.
24. The method of claim 9 , wherein the surface marker is selected from a group consisting of EpCAM, Fas ligand, PD-1, MICA/B, mdr 1, MMPs, CD44, autoreactive antigens, tetraspanins, and MHC class I molecule.