Circulating microrna as a marker for hepatocellular carcinoma

Abstract

Provided herein are methods for the diagnosis, or management of liver diseases, e.g., hepatocellular carcinoma, using profiles of the miRNAs determined from cellular or acellular body fluids.

1 . A method for diagnosing hepatocellular carcinoma (HCC) in a subject, the method comprising: determining, in a body fluid sample from the subject, the level of one or more miRNAs selected from the group consisting of: miR-16, miR-195, and miR-199a, and diagnosing the subject as having hepatocellular carcinoma when a difference in the level of the one or more miRNAs compared to a reference level indicates hepatocellular carcinoma in the subject. 
     
     
         2 . The method of  claim 1 , wherein the body fluid is an acellular body fluid. 
     
     
         3 . The method of  claim 1 , wherein the acellular body fluid is selected from the group consisting of serum and plasma. 
     
     
         4 . The method of  claim 1 , wherein the reference level is the level in a comparable sample from one or more healthy individuals. 
     
     
         5 . The method of  claim 1 , wherein the reference level is the level in a comparable sample from one or more individuals with chronic liver disease. 
     
     
         6 . The method of  claim 1 , wherein the level of one or more miRNAs is compared to a cutoff value determined from the level of one or more miRNAs present in a comparable sample or samples from healthy individuals or individuals with chronic liver disease, and wherein an increase or decrease in the subject value relative to the cutoff value is used to determine a diagnosis for the subject. 
     
     
         7 . The method of  claim 1 , wherein the level of one or more miRNAs is normalized to an internal control RNA level in the sample from the subject. 
     
     
         8 . The method of  claim 7 , wherein the internal control RNA is U6 RNA. 
     
     
         9 . The method of  claim 1 , wherein the one or more miRNAs is miR-16, and a decreased level in the subject compared to a reference level of one or more healthy individuals or individuals with chronic liver disease indicates a diagnosis of HCC in the subject. 
     
     
         10 . The method of  claim 9 , wherein a ΔCt cutoff value of about 6 or greater for miR-16 indicates HCC in the subject, and wherein the ΔCt cutoff value is calculated from the distribution of miR-16 ΔCt values in HCC patients versus the distribution of ΔCt values from a control RNA from a population of chronic liver disease patients. 
     
     
         11 . The method of  claim 1 , wherein the one or more miRNAs is miR-199a, and a decreased level of miR-199a in the subject compared to a reference level of one or more healthy individuals or individuals with chronic liver disease indicates a diagnosis of HCC in the subject. 
     
     
         12 . The method of  claim 11 , wherein a ΔCt cutoff value of about 10 or greater for miR-199a indicates HCC in the subject, wherein the ΔCt cutoff value is calculated from the distribution of miR-199a ΔCt values in HCC patients versus the distribution of ΔCt values from a control RNA from a population of chronic liver disease patients. 
     
     
         13 . The method of  claim 1  further comprising assaying the level of one or more of α-fetoprotein (AFP), AFP-L3%, and des-gamma-carboxyprothrombin (DCP) in the sample. 
     
     
         14 . The method of  claim 13 , wherein the level of miR-16, AFP, AFP-L3%, and DCP are determined in the sample. 
     
     
         15 . A method of diagnosing hepatocellular carcinoma (HCC) in a subject, the method comprising:
 (a) assaying the level of one or more miRNAs selected from the group consisting of: miR-16 and miR-199a in a test sample from a subject;   (b) assaying the level of one or more of AFP, AFP-L3%, and DCP in a test sample from the subject;   (c) determining a score for the subject based on the results obtained in steps (a) and (b); and   (d) comparing the score to a cut-off value in order to determine the presence of HCC in the subject.   
     
     
         16 . The method of  claim 15 , wherein the test sample from step a and b are from the same tissue. 
     
     
         17 . The method of  claim 15 , wherein the test sample from a and b are from different tissues. 
     
     
         18 . The method of  claim 15 , wherein the test ample from a and b comprises hepatic tissue. 
     
     
         19 . The method of  claim 15 , wherein the test sample from a and h is selected from the group consisting of blood, serum, and plasma. 
     
     
         20 . The method of  claim 15 , wherein the level of each of the miR-16, AFP, AFP-L3%, and DCP are assayed. 
     
     
         21 . The method of  claim 15 , wherein the test sample from a and b are one in the same. 
     
     
         22 . The method of  claim 15 , wherein the level of one or more miRNAs is normalized to an internal control RNA level in the sample from the subject. 
     
     
         23 . The method of  claim 22 , wherein the internal control RNA is U6 RNA.