Exosome-associated microrna as a diagnostic marker

Abstract

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.

What is claimed is: 
     
         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.