Human bile contains MicroRNA-laden extracellular vesicles... : Hepatology (original) (raw)

Hepatobiliary Malignancies

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

Li, Ling1,2; Masica, David3; Ishida, Masaharu1; Tomuleasa, Ciprian1,4; Umegaki, Sho5; Kalloo, Anthony N.1; Georgiades, Christos6,7; Singh, Vikesh K.1; Khashab, Mouen1; Amateau, Stuart8; Li, Zhiping1; Okolo, Patrick1; Lennon, Anne-Marie1; Saxena, Payal1; Geschwind, Jean-Francois6; Schlachter, Todd6; Hong, Kelvin6; Pawlik, Timothy M.9; Canto, Marcia1; Law, Joanna1; Sharaiha, Reem10; Weiss, Clifford R.6; Thuluvath, Paul11; Goggins, Michael1; Shin, Eun Ji1; Peng, Haoran1; Kumbhari, Vivek1; Hutfless, Susan1; Zhou, Liya2; Mezey, Esteban1; Meltzer, Stephen J.1; Karchin, Rachel3; Selaru, Florin M.1,12

1_Division of Gastroenterology and Hepatology, Department of Medicine, Beijing, China_

6_Radiology,_

9_Surgery,_

12_Sidney Kimmel Cancer Center, Johns Hopkins Hospital, Baltimore, MD,_

2_Division of Gastroenterology, Third Hospital of Peking University Health Science Center, Beijing, China_

3_Department of Biomedical Engineering and Institute for Computational Medicine Johns Hopkins University, Baltimore, MD,_

4_Center for Genomics and Translational Medicine, Iuliu Hatieganu University of Medicine and Pharmacy, and Department of Hematology, Ion Chiricuta Comprehensive Cancer Center, Cluj Napoca, Romania_

5_Tohoku University School of Medicine, Sendai, Japan_

7_Vascular & Interventional Radiology, American Medical Center, Nicosia, Cyprus_

8_Division of Gastroenterology and Hepatology, University of Colorado, Denver, CO,_

10_Division of Gastroenterology and Hepatology, Weill Cornell Medical College, New York, NY,_

11_The Institute for Digestive Health & Liver Disease at Mercy, Baltimore, MD,_

Received 7 October 2013; Accepted 29 January 2014

Address reprint requests to: Florin M. Selaru, M.D., Johns Hopkins University, 720 Rutland Avenue, Suite 950, Baltimore, MD 21205. E-mail: [email protected]; fax: 410-614-9612.

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

Abstract

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)