Delineating the Metabolic Phenotype of Biopsy-Derived Kidney Cells

Masataka Kawamura; Catherine Parmentier; Lisa A. Robinson; Markus Selzner; Caitriona M. McEvoy; Julia M. Murphy; Maya G. Bozzo-Rey; Sarah Q. Crome; Ana Konvalinka; Sergi Clotet-Freixas

doi:10.3791/65457

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Method Article

Delineating the Metabolic Phenotype of Biopsy-Derived Kidney Cells

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DOI:

10.3791/65457

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June 17th, 2025

Masataka Kawamura¹^,²^,³ , Catherine Parmentier¹^,³ , Lisa A. Robinson²^,⁴^,⁵ , Markus Selzner¹^,³ , Caitriona M. McEvoy⁶^,⁷^,⁸ , Julia M. Murphy¹^,⁶^,⁹ , Maya G. Bozzo-Rey¹⁰ , Sarah Q. Crome¹^,⁶^,⁹ , Ana Konvalinka¹^,⁴^,⁶^,¹¹^,¹² , Sergi Clotet-Freixas¹⁰

¹Soham and Shaila Ajmera Family Transplant Centre, University Health Network, ²Division of Nephrology, The Hospital for Sick Children, ³Division of General Surgery, University Health Network, ⁴Institute of Medical Science, University of Toronto, ⁵Program in Cell Biology, The Hospital for Sick Children Research Institute, ⁶Toronto General Hospital Research Institute, University Health Network, ⁷Division of Nephrology, Tallaght University Hospital, ⁸Trinity Kidney Centre, Trinity College Dublin, ⁹Department of Immunology, University of Toronto, ¹⁰Division of Nephrology, McMaster University and St. Joseph's Healthcare, ¹¹Division of Nephrology, Department of Medicine, University Health Network, ¹²Department of Laboratory Medicine and Pathobiology, University of Toronto

Summary

This protocol describes a unique method to conduct metabolic profiling of biopsy-derived kidney cells. The approach presented was optimized in healthy adult male pigs and has the potential to enhance the real-time assessment of kidney allograft viability. This method may also help uncover new biology across multiple metabolism-based kidney diseases.

Abstract

Kidney transplantation is the optimal treatment for end-stage kidney disease; however, transplanted kidneys are often lost prematurely, with up to 50% graft loss at 10 years post-transplant. One of the major causes of premature graft loss is the injury sustained by the graft at the time of transplantation, known as ischemia-reperfusion injury (IRI). Delayed graft function (DGF), defined as the need for dialysis in the first week post-transplant, is a manifestation of severe IRI that shows functional and histologic features of acute kidney injury (AKI). While the mechanisms driving AKI remain unclear, accumulating evidence suggests that altered metabolic function in the allograft mediates AKI and may be the reason for DGF. Thus, deciphering and monitoring the metabolic underpinnings of IRI will improve our capacity to diagnose and prevent AKI. This article describes a unique method to assess mitochondrial respiration (by means of oxygen consumption rate), glycolysis (extracellular acidification rate), and intracellular ATP levels in needle biopsy-derived kidney cell suspensions. The methodology has been optimized in healthy adult male pigs and validated in a porcine model of auto-transplantation. The approach presented has the potential to enhance the real-time assessment of kidney allograft viability in the clinic. Profiling metabolism in patient-derived biopsies may also uncover new biology in other metabolism-based kidney diseases.

Introduction

Kidney transplantation is the optimal treatment for end-stage kidney disease¹^,²^,³^,⁴; however, transplanted kidneys are often lost prematurely, with up to 50% graft loss at 10 years post-transplant⁵. Affected patients have increased morbidity and mortality and pose a major economic burden on healthcare systems⁶. A major cause of premature graft loss is the injury sustained by the graft at the time of transplantation, known as ischemia-reperfusion injury (IRI). IRI is an unavoidable injury caused....