Exosome-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 RNAs present in cancer-derived exosomes isolated from a biological sample from the subject.
1. A method for evaluating treatment efficacy and/or progression of a cancer in a subject, comprising:
(a) isolating cancer-derived microvesicles from a biological sample of a subject;
(b) determining an amount of one or more microRNAs in the isolated cancer-derived microvesicles; and
(c) determining any measurable change in the amounts of the one or more microRNAs to thereby evaluate treatment efficacy and/or progression of the cancer in the subject.
2. The method of claim 1 , wherein the cancer is selected from the group consisting of ovarian cancer, cervical cancer, breast cancer, endometrial cancer, colon cancer, prostate cancer, lung cancer, melanoma, and pancreatic cancer.
3. The method of claim 1 , wherein the subject is human.
4. The method of claim 1 , wherein the biological sample comprises a first biological sample collected prior to initiation of treatment for the cancer and/or onset of the cancer and a second biological sample from the subject collected after initiation of the treatment or onset.
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 cancer-derived microvesicles comprises using size exclusion chromatography to isolate the cancer-derived microvesicles.
7. The method of claim 6 , wherein isolating the cancer-derived microvesicles further comprises centrifuging a chromatography fraction comprising the cancer-derived microvesicles.
8. The method of claim 7 , wherein the chromatography fraction is a void volume fraction.
9. The method of claim 6 , wherein the cancer-derived microvesicles are separated from non-cancer-derived microvesicles by immunosorbent capture using an antibody to a cell surface antigen.
10. The method of claim 9 , wherein the antibody is an anti-epithelial cell adhesion molecule (anti-EpCAM) antibody.
11. The method of claim 1 , wherein determining the amount of the one or more microRNAs comprises labeling the one or more microRNAs.
12. The method of claim 1 , wherein determining the amount of the one or more microRNAs comprises capturing the one or more microRNAs with one or more polynucleotide probes that each selectively bind the one or more microRNAs.
13. The method of claim 1 , wherein determining the amount of the one or more microRNAs comprises using a real-time polymerase chain reaction to quantitate the amount of the one or more microRNAs.
14. The method of claim 1 , wherein the one or more microRNAs are miR-200c and miR-214.
15. The method of claim 1 , further comprising selecting a treatment or modifying a treatment for the cancer based on the amount of the one or more microRNAs determined.
16. The method of claim 1 , wherein the one or more microRNAs are selected from the group consisting of miR-21, miR-141, miR-200a, miR-200b, miR-200c, miR-203, miR-205, and miR-214.