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


CONCLUSIONS
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.
Division of Gastrenterology and Hepatology, Department of Medicine, Johns Hopkins Hospital, Baltimore, Maryland; USA.
Ling
L
Li
David
D
Masica
Masaharu
M
Ishida
Ciprian
C
Tomuleasa
Sho
S
Umegaki
Anthony N
AN
Kalloo
Christos
C
Georgiades
Vikesh K
VK
Singh
Mouen
M
Khashab
Stuart
S
Amateau
Zhiping
Z
Li
Patrick
P
Okolo
Anne-Marie
AM
Lennon
Payal
P
Saxena
Jean-Francois
JF
Geschwind
Todd
T
Schlachter
Kelvin
K
Hong
Timothy M
TM
Pawlik
Marcia
M
Canto
Joanna
J
Law
Reem
R
Sharaiha
Clifford R
CR
Weiss
Paul
P
Thuluvath
Michael
M
Goggins
Eun Ji
EJ
Shin
Haoran
H
Peng
Vivek
V
Kumbhari
Susan
S
Hutfless
Liya
L
Zhou
Esteban
E
Mezey
Stephen J
SJ
Meltzer
Rachel
R
Karchin
Florin M
FM
Selaru
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001646:Bile / Q000737:chemistry*; D001650:Bile Duct Neoplasms / Q000175:diagnosis* / Q000378:metabolism; D001653:Bile Ducts, Intrahepatic; D015415:Biomarkers / Q000032:analysis; D016022:Case-Control Studies; D018281:Cholangiocarcinoma / Q000175:diagnosis* / Q000378:metabolism; D022162:Cytoplasmic Vesicles / Q000737:chemistry / Q000378:metabolism; D005260:Female; D006801:Humans; D008297:Male; D035683:MicroRNAs / Q000032:analysis* / Q000378:metabolism; D008875:Middle Aged; D060388:Support Vector Machine; D055815:Young Adult
D015415:Biomarkers; D035683:MicroRNAs