Multiplex PCR Screening of MicroRNAs in Graft Preservation Fluid during Liver Transplantation for Biomarker Discovery

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J.W. Selten, H.P. Roest, A.J.M. Gillis, L.C.J. Dorssers, J. de Jonge, L.H.J. Looijenga, J.N.M Ijzermans, L.J.W. van der Laan

Chair(s): Drs. K.C.A. Geneugelijk & prof. dr. C. van Kooten

Thursday 9 march 2017

15:18 - 15:30h at Hendrik Marsmanzaal

Categories: Parallel - Basaal

Parallel session: Parallelsessie XV - Basaal - Moleculaire analyses en het adaptieve immuunsysteem in orgaantransplantatie

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Introduction:
MicroRNAs (miRNAs) have been extensively investigated in recent years as biomarkers in liver transplantation. A variety of miRNAs in serum, tissue and bile have been demonstrated to correlate to rejection, early allograft dysfunction (EAD) and biliary complications after transplantation. However, the global miRNAs profiles in graft perfusion fluids during liver preservation have not been reported. In this study we aimed to identify perfusate miRNA profiles and investigate their potential to predict graft outcomes after transplantation.

Material & methods:
Cell-free preservation fluids of 32 liver grafts at the end of cold storage were analyzed for miRNA content using Taqman microRNA array card A. 50% of grafts were from donation after brain death (DBD) and 50% from donation after circulatory death (DCD). For both donor types 8 grafts resulting in EAD and 8 resulting in non-EAD were included. Bioinformatics analysis was performed using the R-package HTqPCR on Ath miR-159a normalized datasets.

Results:
220 miRNAs were reliably detectable in perfusates. A difference in miRNA levels was seen between DCD and DBD livers for miR-523-3p, miR-525-5p, miR-382, miR-7a and miR-200a (p<0.01). Furthermore, 11 miRNAs were identified as significantly different between EAD and non-EAD grafts (miR-491-5p, miR-200c, miR-382, miR-220, miR-221, miR-510, miR-542, miR-518b, miR-379 miR-204 and miR-122, p<0.01), known to be liver abundant. Perfusates of liver grafts which developed biliary lesions after liver transplantation showed six new miRNAs (miR-455-5p, miR-191, miR-324-5p, miR-142, miR-302, miR-410, p<0.01).

Conclusion:
In this discovery study we have identified several new miRNAs in graft preservation solutions of liver grafts related to donor type and post transplantation graft function. Further research is ongoing to validate the use of these miRNAs to assess graft quality during preservation in a larger cohort.