Circulating biomarkers for metastatic prostate cancer
Biomarkers can be assessed for diagnostic, therapy-related or prognostic methods to identify phenotypes, such as a condition or disease, or the stage or progression of a disease. Circulating biomarkers from a bodily fluid can be used in profiling of physiological states or determining phenotypes. These include nucleic acids, protein, and circulating structures such as vesicles. Biomarkers can be used for theranostic purposes to select candidate treatment regimens for diseases, conditions, disease stages, and stages of a condition, and can also be used to determine treatment efficacy. The biomarkers can be circulating biomarkers, including vesicles and microRNA.
1. A method of distinguishing metastatic prostate cancer from non-metastatic prostate cancer in a human patient comprising:
a) isolating at least one microvesicle from a bodily fluid sample from the human patient by contacting the sample with at least one binding agent to one or more microvesicle surface antigens;
b) isolating microRNA from the at least one microvesicle isolated in a);
c) detecting the level of a plurality of microRNA within the microRNA isolated in b), wherein the plurality of microRNA comprises hsa-miR-574-3p (miR-574-3p), hsa-miR-200b (miR-200b), hsa-miR-375 (miR-375), hsa-miR-141 (miR-141), hsa-miR-331-3p (miR-331-3p) and hsa-miR-181a (miR-181a);
d) comparing the level of each of the plurality of microRNA detected in c) to a reference; and
e) classifying the sample as metastatic prostate cancer or non-metastatic prostate cancer based on the comparison in d).
2. The method of claim 1 , wherein the plurality of microRNA further comprises at least one of hsa-miR-100, hsa-miR-1236, hsa-miR-1296, hsa-miR-146b-5p, hsa-miR-17*, hsa-miR-20a*, hsa-miR-23a*, hsa-miR-452, hsa-miR-572, hsa-miR-577, hsa-miR-582-3p, hsa-miR-937, miR-10a, miR-134, miR-30a, miR-32, miR-495, miR-564, miR-570, and miR-885-3p.
3. The method of claim 1 , wherein the plurality of microRNA further comprises at least one of miR-495, miR-10a, miR-30a, miR-570, miR-32, miR-885-3p, miR-564, and miR-134.
4. The method of claim 1 , wherein the plurality of microRNA further comprises at least one of hsa-miR-582-3p, hsa-miR-17*, hsa-miR-1296, hsa-miR-20a*, hsa-miR-100, hsa-miR-452, and hsa-miR-577.
5. The method of claim 1 , wherein the plurality of microRNA further comprises at least one of hsa-miR-452, hsa-miR-146b-5p, hsa-miR-1296, hsa-miR-17*, hsa-miR-100, hsa-miR-20a*, hsa-miR-572, hsa-miR-1236, hsa-miR-937, and hsa-miR-23a*.
6. The method of claim 1 , wherein the step of comparing the level of the plurality of microRNA to the reference comprises determining whether any member of the plurality of microRNA is altered relative to the reference.
7. The method of claim 1 , wherein the bodily fluid comprises peripheral blood, sera, plasma, ascites, urine, cerebrospinal fluid (CSF), sputum, saliva, bone marrow, synovial fluid, aqueous humor, cerumen, broncheoalveolar lavage fluid, semen, prostatic fluid, cowper's fluid or pre-ejaculatory fluid, sweat, fecal matter, tears, cyst fluid, pleural fluid, peritoneal fluid, pericardial fluid, lymph, chyme, chyle, bile, interstitial fluid, pus, sebum, vomit, mucosal secretion, stool water, pancreatic juice, lavage fluids from sinus cavities, bronchopulmonary aspirates, blastocyl cavity fluid, or a combination thereof.
8. The method of claim 1 , wherein the bodily fluid comprises urine, whole blood, plasma or sera.
9. The method of claim 1 , wherein the at least one microvesicle has a diameter between 20 nm and 800 nm.
10. The method of claim 1 , wherein the at least one microvesicle has a diameter between 20 nm and 200 nm.
11. The method of claim 1 , wherein isolating the at least one microvesicle further comprises size exclusion chromatography, density gradient centrifugation, differential centrifugation, nanomembrane ultrafiltration, immunoabsorbent capture, affinity purification, affinity capture, immunoassay, microfluidic separation, flow cytometry or combinations thereof.
12. The method of claim 1 , wherein the at least one binding agent comprises an antibody, antibody fragment, aptamer, or a combination thereof.
13. The method of claim 1 , wherein the at least one surface antigen comprises at least one of CD9, CD63, CD81, PSMA, PCSA, B7H3 and EpCam.
14. The method of claim 1 , wherein the at least one surface antigen comprises at least one of a tetraspanin, CD9, CD63, CD81, CD63, CD9, CD81, CD82, CD37, CD53, Rab-5b, Annexin V, MFG-E8, or a protein in Table 3.
15. The method of claim 1 , wherein the at least one surface antigen comprises at least one of CD9, PSMA, PCSA, CD63, CD81, B7H3, IL 6, OPG-13, IL6R, PA2G4, EZH2, RUNX2, SERPINB3, and EpCam.