MicroRNA profiles in graft preservation solution are predictive of ischemic-type biliary lesions after liver transplantation.
OBJECTIVE
Ischemic-type biliary lesions (ITBL) are the second most common cause of graft loss after liver transplantation. Though the exact pathophysiology of ITBL is unknown, bile duct injury during graft preservation is considered to be a major cause. Here we investigated whether the release of cholangiocyte-derived microRNAs (CDmiRs) during graft preservation is predictive of the development of ITBL after liver transplantation.


METHODS
Graft preservation solutions (perfusates) and paired liver biopsies collected at the end of cold ischemia were analysed by RT-qPCR for CDmiR-30e, CDmiR-222, and CDmiR-296 and hepatocyte-derived miRNAs (HDmiRs) HDmiR-122 and HDmiR-148a. MicroRNAs in perfusates were evaluated on their stability by incubation and fractionation experiments. MicroRNA profiles in perfusates from grafts that developed ITBL (n=20) and grafts without biliary strictures (n=37) were compared.


RESULTS
MicroRNAs in perfusates were proven to be stable and protected against degradation by interacting proteins. Ratios between HDmiRs/CDmiRs were significantly higher in perfusates obtained from grafts that developed ITBL (p<0.01) and were identified as an independent risk factor by multivariate analysis (p<0.01, HR: 6.89). The discriminative power of HDmiRs/CDmiRs in perfusates was validated by analysis of separate brain death- (DBD) and cardiac death donors (DBD; p ≤ 0.016) and was superior to expression in liver biopsies (C=0.77 in perfusates vs. C<0.50 in biopsies).


CONCLUSIONS
This study demonstrates that differential release of CDmiRs during graft preservation is predictive of the development of ITBL after liver transplantation. This provides new evidence for the link between graft-related bile duct injury and the risk for later development of ITBL.
Department of Surgery, Erasmus MC-University Medical Center, Rotterdam, The Netherlands.
Cornelia J
CJ
Verhoeven
Waqar R R
WR
Farid
Petra E
PE
de Ruiter
Bettina E
BE
Hansen
Henk P
HP
Roest
Jeroen
J
de Jonge
Jaap
J
Kwekkeboom
Herold J
HJ
Metselaar
Hugo W
HW
Tilanus
Geert
G
Kazemier
Luc J W
LJ
van der Laan
D000328:Adult; D000368:Aged; D001652:Bile Ducts / Q000098:blood supply*; D005260:Female; D006801:Humans; D007511:Ischemia / Q000209:etiology*; D016031:Liver Transplantation / Q000009:adverse effects*; D008297:Male; D035683:MicroRNAs / Q000032:analysis*; D008875:Middle Aged; D019188:Organ Preservation Solutions / Q000032:analysis*
D035683:MicroRNAs; D019188:Organ Preservation Solutions
ALT; AST; Biomarker; CDmiRs; CMV; Caenorhabditis elegans miRNA; Cholangiocyte; DCD; ERCP; Graft survival; HAT; HDmiRs; HTK; ITBL; Marginal donors; Non-anastomotic biliary strictures; PNF; PSC; PTC; RNA; RT-PCR; UW; University of Wisconsin solution; alanine aminotransferase; aspartate aminotransferase; cDNA; cel-miR; cholangiocyte-derived miRNAs; complement deoxyribonucleic acid; cytomegalovirus; donation after cardiac death; endoscopic retrograde cholangiopancreatography; hepatic artery thrombosis; hepatocyte-derived miRNAs; histidine-tryptophan-ketoglutarate solution; ischemic-type biliary lesions; miRNAs; microRNAs; percutaneous transhepatic cholangiography; primary non-function; primary sclerosing cholangitis; real time polymerase chain reaction; ribonucleic acid