Subclinical Changes in Deceased Donor Kidney Proteomes are Associated with 12-month Allograft Function Posttransplantation – a Preliminary Study

Background Cerebral injury during Donation after Brain Death (DBD) may induce systemic damage affecting long-term kidney function posttransplantation. Conventional evaluation of donor organ quality as a triage for transplantation is of limited utility. Methods We compared donor kidneys yielding opposing extremes of the continuum of posttransplantation outcomes by several common kidney biopsy evaluation techniques including Kidney Donor Profile Index (KDPI) and Remuzzi scoring, and analysed tissue from a minimal sample cohort using Label-Free Quantitation (LFQ) mass spectrometry. Further assessment of the proteomic results was performed by orthogonal quantitative comparisons of selected key proteins by immunoblotting. Results We show that common evaluation techniques of kidney biopsies were not predictive for posttransplantation outcomes. In contrast, despite the limited cohort size, the proteomic analysis was able to clearly differentiate between kidneys yielding extreme posttransplantation outcome differences. Pathway analysis of the proteomic data suggested that outcome-related variance in protein abundance associated with profibrotic, apoptosis and antioxidant proteins. Immunoblotting confirmation further supported this observation. Conclusions We present preliminary data indicating that there is scope for existing evaluation approaches to be supplemented by the analysis of proteomic differences. Furthermore, the observed outcome-related variance in a limited cohort was supported by immunoblotting and is consistent with mechanisms previously implicated in the development of injury and cytoprotection in kidney transplantation. This is an open access article distributed under the Creative Commons Attribution License 4.0 (CCBY), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. + Equal contribution ◊ Equal contribution Corresponding Author: Dr. Maria Kaisar (maria.kaisar@nds.ox.ac.uk), Nuffield Department of Surgical Sciences, University of Oxford, Oxford, OX3 7LJ, United Kingdom AUTHORSHIP PAGE Authorship: MK, BMK and RJP designed this study. MK, AZ, NW as members of the National Management Team and the QUOD Steering Committee participated in the establishment of the QUOD biobank. RJP is the Coordinator of the QUOD consortium. Clinical evaluation of samples was performed by MK. Sample preparation for proteomic study was performed by MK, MLT and HH. Mass spectrometric analysis was performed by MLT and HH. Mass spectrometric data was processed and searched by MK and MLT. Statistical analysis of proteomic quantitation was performed by MK and PDC. Immunoblotting was performed by LvD. Remuzzi scoring was performed by AK. The paper was written by MK, PDC, BMK and RJP with input from the other authors. Disclosure: The authors declare no conflicts of interest Funding: This work was supported by research funding from NHS Blood and Transplant Trust Fund TF031 and Oxford Transplant Foundation award to M.K., a John Fell Fund 133/075 and Wellcome Trust grant 097813/Z/11/Z to B.M.K. and COPE FP7 grant award to R.J.P. Copyright © 2018 Wolters Kluwer Health, Inc. All rights reserved.

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