TY - JOUR T1 - Normothermic Ex-vivo Kidney Perfusion in a Porcine Auto-Transplantation Model Preserves the Expression of Key Mitochondrial Proteins: An Unbiased Proteomics Analysis JF - bioRxiv DO - 10.1101/2020.08.17.253252 SP - 2020.08.17.253252 AU - Caitriona M. McEvoy AU - Sergi Clotet-Freixas AU - Tomas Tokar AU - Chiara Pastrello AU - Shelby Reid AU - Ihor Batruch AU - Adrien A.E. RaoPeters AU - J. Moritz Kaths AU - Peter Urbanellis AU - Sofia Farkona AU - Julie A.D. Van AU - Bradley L. Urquhart AU - Rohan John AU - Igor Jurisica AU - Lisa A. Robinson AU - Markus Selzner AU - Ana Konvalinka Y1 - 2020/01/01 UR - http://biorxiv.org/content/early/2020/08/18/2020.08.17.253252.abstract N2 - Normothermic ex-vivo kidney perfusion (NEVKP) results in significantly improved graft function in porcine auto-transplant models of DCD injury compared to static cold storage (SCS); however, the molecular mechanisms underlying these beneficial effects remain unclear. We performed an unbiased proteomics analysis of 28 kidney biopsies obtained at 3 time points from pig kidneys subjected to 30-minutes of warm ischemia, followed by 8 hours of NEVKP or SCS, and auto-transplantation. 70/6593 proteins quantified were differentially expressed between NEVKP and SCS groups (FDR<0.05). Proteins increased in NEVKP mediated key metabolic processes including fatty acid ß-oxidation, the TCA-cycle and oxidative phosphorylation. Comparison of our findings with external datasets of ischemia-reperfusion, and other models of kidney injury confirmed that 47 of our proteins represent a common signature of kidney injury reversed or attenuated by NEVKP. We validated key metabolic proteins (ETFB, CPT2) by immunoblotting. Transcription factor databases identified PPARGC1A, PPARA/G/D and RXRA/B as the upstream regulators of our dataset, and we confirmed their increased expression in NEVKP with RT-PCR. The proteome-level changes observed in NEVKP mediate critical metabolic pathways that may explain the improved graft function observed. These effects may be coordinated by PPAR-family transcription factors, and may represent novel therapeutic targets in ischemia-reperfusion injury.Competing Interest StatementThe authors have declared no competing interest. ER -