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 of detecting a plurality of miRNAs in a biological sample, comprising:
i) isolating exosomes from the biological sample;
ii) isolating miRNA from the isolated exosomes; and
iii) determining the amount of the plurality of miRNAs in the isolated miRNA, wherein the plurality of miRNAs comprises miR-17-3p, miR-21, miR-106a, miR-146, miR-155, miR-191, miR-192, miR-203, miR-205, miR-210, miR-212, and miR-214, thereby detecting the plurality of miRNAs in the biological sample.
2 . The method of claim 1 , wherein the biological sample is from a human subject.
3 . The method of claim 2 , wherein the subject has a cancer.
4 . The method of claim 3 , wherein the cancer comprises lung cancer.
5 . 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.
6 . The method of claim 1 , wherein isolating the exosomes comprises using size exclusion chromatography.
7 . The method of claim 6 , wherein isolating the exosomes further comprises centrifuging a chromatography fraction comprising the exosomes.
8 . The method of claim 7 , wherein the chromatography fraction is a void volume fraction.
9 . The method of claim 1 , wherein isolating the exosomes comprises using immunosorbent capture.
10 . The method of claim 9 , wherein the immunosorbent capture comprises using antibodies to capture the exosomes.
11 . The method of claim 10 , wherein the antibodies comprise antibodies to at least one of EpCAM, Fas ligand, PD-1, MICA/B, mdr1, an MMP, CD44, an autoreactive antigen, a tetraspanin, and an MHC class I molecule.
12 . The method of claim 1 , wherein determining the amount of the plurality of miRNAs comprises labeling the members of the plurality of miRNAs.
13 . The method of claim 1 , wherein determining the amount of the plurality of miRNAs comprises capturing members of the plurality of miRNAs with a plurality of polynucleotide probes that each selectively bind members of the plurality of miRNAs.
14 . The method of claim 1 , wherein determining the amount of the plurality of miRNAs comprises using a real-time polymerase chain reaction or a microarray to quantitate the amount of the members of the plurality of miRNAs.
15 . The method of claim 1 , further comprising providing a diagnosis or prognosis of a cancer based on the amount of the members of the plurality of miRNAs determined.
16 . The method of claim 1 , further comprising selecting a treatment or modifying a treatment for a cancer based on the amount of the members of the plurality of miRNAs determined.
17 . The method of claim 15 , wherein the cancer comprises lung cancer.
18 . The method of claim 16 , wherein the cancer comprises lung cancer.