Human bile contains MicroRNA-laden extracellular vesicles that can be used for cholangiocarcinoma diagnosis



This article is corrected by:

  1. Errata: Correction Volume 60, Issue 6, 2135, Article first published online: 24 November 2014

  • Potential conflict of interest: Dr. Kaloo owns stock and has intellectual property rights in Apollo. He consults for Checkmed and Pentax. Dr. Saxena consults for and received grants from Boston Scientific and also received grants from Cook Medical. Dr. Geschwind consults for and received grants from Biocompatibles/BTG, Bayer, Guerbet, Nordion/BTG, and Phillips. He consults for Jennerex and received grants from Theshold. He is the founder and CEO of PreScience Labs, LLC. Dr. Thuluvath advises, is on the speakers' bureau for, and received grants from Vertex and Gilead. He advises Janssen and is on the speakers' bureau for Onyx. He received grants from Boehringer Ingelheim, Novartis, Bristol-Myers Squibb, Eisai, and Salix.

  • This study was supported by a K08 Award (DK090154-01) from the National Institutes of Health (NIH; to F.M.S.) and by an Early Research and Detection Network (EDRN) Associate Membership supported by an U01 Award (CA086402) from the NIH. Dr. Meltzer is an American Cancer Society Clinical Research Professor.

  • See Editorial on Page 782


Cholangiocarcinoma (CCA) presents significant diagnostic challenges, resulting in late patient diagnosis and poor survival rates. Primary sclerosing cholangitis (PSC) patients pose a particularly difficult clinical dilemma because they harbor chronic biliary strictures that are difficult to distinguish from CCA. MicroRNAs (miRs) have recently emerged as a valuable class of diagnostic markers; however, thus far, neither extracellular vesicles (EVs) nor miRs within EVs have been investigated in human bile. We aimed to comprehensively characterize human biliary EVs, including their miR content. We have established the presence of extracellular vesicles in human bile. In addition, we have demonstrated that human biliary EVs contain abundant miR species, which are stable and therefore amenable to the development of disease marker panels. Furthermore, we have characterized the protein content, size, numbers, and size distribution of human biliary EVs. Utilizing multivariate organization of combinatorial alterations (MOCA), we defined a novel biliary vesicle miR-based panel for CCA diagnosis that demonstrated a sensitivity of 67% and specificity of 96%. Importantly, our control group contained 13 PSC patients, 16 with biliary obstruction of varying etiologies (including benign biliary stricture, papillary stenosis, choledocholithiasis, extrinsic compression from pancreatic cysts, and cholangitis), and 3 with bile leak syndromes. Clinically, these types of patients present with a biliary obstructive clinical picture that could be confused with CCA. Conclusion: These findings establish the importance of using extracellular vesicles, rather than whole bile, for developing miR-based disease markers in bile. Finally, we report on the development of a novel bile-based CCA diagnostic panel that is stable, reproducible, and has potential clinical utility. (Hepatology 2014;60:896–907)