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A Proteomic Analysis of Kidneys Subjected to Normothermic Ex Vivo Kidney Perfusion (NEVKP) Demonstrates That Metabolism is an Important Determinant of Kidney Function Following Warm Ischemia

A. Konvalinka,1,4 T. Tokar,2 S. Reid,4 P. Urbanellis,4 M. Hamar,4 M. Kaths,4 I. Jurisica,2 L. Robinson,4,3 M. Selzner.4,5

1Medicine/Nephrology, University Health Network, Toronto, ON, Canada
2Medical Biophysics/Computer Science, University of Toronto, Toronto, ON, Canada
3Pediatrics/Nephrology, Hospital for Sick Children, Toronto, ON, Canada
4Institute of Medical Science, University of Toronto, Toronto, ON, Canada
5Surgery/General Surgery, University Health Network, Toronto, ON, Canada.

Meeting: 2018 American Transplant Congress

Abstract number: B17

Keywords: Kidney transplantation, Pig, Preservation, Warm ischemia

Session Information

Session Name: Poster Session B: Acute and Chronic Graft Injury

Session Type: Poster Session

Date: Sunday, June 3, 2018

Session Time: 6:00pm-7:00pm

 Presentation Time: 6:00pm-7:00pm

Location: Hall 4EF

Kidney transplantation reduces mortality compared to staying on dialysis, incentivizing the use of marginal kidneys in face of organ shortage. These marginal kidneys after cardiocirculatory death are damaged by warm ischemia and have inferior outcomes. Graft repair prior to transplant would increase the number of available kidneys and improve recipient outcomes. We developed a novel NEVKP system in a porcine kidney transplant model. Eight hours of NEVKP compared to static cold storage (SCS) improved graft function following transplantation. However, the molecular mechanisms responsible for the beneficial effect of NEVKP are not known.

Porcine kidneys were subjected to 30 minutes of warm ischemia followed by 8 hours of NEVKP or SCS, with subsequent auto-transplantation and nephrectomy of the contralateral kidney. Serum creatinine was measured daily, until POD3. Kidney biopsies were taken at time of explant, 30 minutes post-reperfusion and on POD3 (n=30 biopsies). Biopsy tissue was subjected to unbiased proteomic analysis.

We quantified 6354 proteins. Serum creatinine was significantly (p<2.23e-15) lower in NEVKP compared to SCS kidneys, with the largest difference on POD3. Principal component analysis segregated POD3 biopsy proteins from proteins at other time points. 69 proteins were differentially expressed (p<0.05; 2-way ANOVA/Tukey's HSD) between NEVKP and SCS groups and across 3 time points. Proteins increased in SCS compared to NEVKP kidneys at POD3 mediate fibrosis, oxidative stress and inflammation (THBS1, XRCC6, ALG2, LIPA, CLPTM1L, HPCAL1). At 30 minutes post-reperfusion, CYP1A1 was found exclusively in NEVKP biopsies. CYP1A1 is a metabolic protein previously associated with protection against ischemic injury. Metabolism was a key enriched biological process.

NEVKP significantly improved kidney function compared to SCS. We have identified groups of proteins that may be involved in the beneficial effects of NEVKP. Further studies will delineate the function of these proteins in ischemia reperfusion injury.

CITATION INFORMATION: Konvalinka A., Tokar T., Reid S., Urbanellis P., Hamar M., Kaths M., Jurisica I., Robinson L., Selzner M. A Proteomic Analysis of Kidneys Subjected to Normothermic Ex Vivo Kidney Perfusion (NEVKP) Demonstrates That Metabolism is an Important Determinant of Kidney Function Following Warm Ischemia Am J Transplant. 2017;17 (suppl 3).

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To cite this abstract in AMA style:

Konvalinka A, Tokar T, Reid S, Urbanellis P, Hamar M, Kaths M, Jurisica I, Robinson L, Selzner M. A Proteomic Analysis of Kidneys Subjected to Normothermic Ex Vivo Kidney Perfusion (NEVKP) Demonstrates That Metabolism is an Important Determinant of Kidney Function Following Warm Ischemia [abstract]. https://atcmeetingabstracts.com/abstract/a-proteomic-analysis-of-kidneys-subjected-to-normothermic-ex-vivo-kidney-perfusion-nevkp-demonstrates-that-metabolism-is-an-important-determinant-of-kidney-function-following-warm-ischemia/. Accessed June 6, 2025.

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