EXOSOME-ASSOCIATED MICRORNA AS A DIAGNOSTIC MARKER
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 biomarkers from microvesicles, comprising isolating a population of extracellular microvesicles from a biological sample using a microvesicle surface marker, isolating microRNA from said population of microvesicles, and determining a presence of one or more microRNA in said isolated microRNA.
2 . The method of claim 1 , wherein the surface marker is selected from a group consisting of EpCAM, Fas ligand, PD-1, MICA/B, mdr1, MMPs, CD44, autoreactive antigens, a tetraspanin, and an MHC class I molecule.
3 . The method of claim 1 , wherein the microRNA is selected from miRNA set forth in Table 1.
4 . The method of claim 1 , wherein the microRNA comprises one or more microRNA selected from the group consisting of miR-016, miR-019b, miR-020a, miR-21, miR-024, miR-026a, miR-32, miR-126, miR-141, miR-146a, miR-146b, miR-191, miR-200a, miR-200b, miR-200c, miR-203, miR-205, miR-214, miR-222, miR-223, and miR-484.
5 . A method of determining a microRNA expression profile of microvesicles, comprising: isolating a population of extracellular microvesicles from a biological liquid sample; isolating microRNA from said isolated microvesicles and determining an expression profile of one or more microRNA in said isolated microRNA; and
comparing the expression profile with a profile from a reference sample to determine the microRNA expression profile for said microvesicles.
6 . The method of claim 5 , wherein the biological liquid sample comprises milk, blood, serum, plasma, ascites, cyst fluid, pleural fluid, peritoneal fluid, cerebral spinal fluid, tears, urine, saliva, sputum, or combinations thereof.
7 . The method of claim 5 , wherein said determining step comprises labeling the one or more microRNA with a detectable label.
8 . The method of claim 5 , wherein said determining comprises capturing the one or more microRNA with one or more polynucleotide probe that selectively binds each of the one or more microRNA.
9 . The method of claim 5 , wherein said determining comprises using a real-time polymerase chain reaction or a nucleic acid array.
10 . The method of claim 5 , wherein isolating the microvesicle population comprises using size exclusion chromatography, filtration or immunosorbent capture.
11 . The method of claim 5 , wherein the microRNA is selected from miRNA set forth in Table 1.
12 . The method of claim 5 , wherein the microRNA is selected from miRNA set forth in Table 1. The method of claim 1 or 2 , wherein the microRNA comprises one or more microRNA selected from the group consisting of miR-016, miR-019b, miR-020a, miR-21, miR-024, miR-026a, miR-32, miR-126, miR-141, miR-146a, miR-146b, miR-191, miR-200a, miR-200b, miR-200c, miR-203, miR-205, miR-214, miR-222, miR-223, and miR-484.