The impact of normothermic and hypothermic preservation methods on kidney lipidome-comparative study using chemical biopsy with microextraction probes

Natalia Warmuzińska; Kamil Łuczykowski; Iga Stryjak; Hernando Rosales-Solano; Peter Urbanellis; Janusz Pawliszyn; Markus Selzner; Barbara Bojko

doi:10.3389/fmolb.2024.1341108

The impact of normothermic and hypothermic preservation methods on kidney lipidome-comparative study using chemical biopsy with microextraction probes

Front Mol Biosci. 2024 May 9:11:1341108. doi: 10.3389/fmolb.2024.1341108. eCollection 2024.

Authors

Natalia Warmuzińska¹, Kamil Łuczykowski¹, Iga Stryjak¹, Hernando Rosales-Solano², Peter Urbanellis³, Janusz Pawliszyn², Markus Selzner^{3

4}, Barbara Bojko¹

Affiliations

¹ Department of Pharmacodynamics and Molecular Pharmacology, Faculty of Pharmacy, Nicolaus Copernicus University in Torun, Collegium Medicum in Bydgoszcz, Bydgoszcz, Poland.
² Department of Chemistry, University of Waterloo, Waterloo, ON, Canada.
³ Ajmera Transplant Center, Department of Surgery, Toronto General Hospital, University Health Network, Toronto, ON, Canada.
⁴ Department of Medicine, Toronto General Hospital, Toronto, ON, Canada.

Abstract

Introduction: Normothermic ex vivo kidney perfusion (NEVKP) is designed to replicate physiological conditions to improve graft outcomes. A comparison of the impact of hypothermic and normothermic preservation techniques on graft quality was performed by lipidomic profiling using solid-phase microextraction (SPME) chemical biopsy as a minimally invasive sampling approach.

Methods: Direct kidney sampling was conducted using SPME probes coated with a mixed-mode extraction phase in a porcine autotransplantation model of the renal donor after cardiac death, comparing three preservation methods: static cold storage (SCS), NEVKP, and hypothermic machine perfusion (HMP). The lipidomic analysis was done using ultra-high-performance liquid chromatography coupled with a Q-Exactive Focus Orbitrap mass spectrometer.

Results: Chemometric analysis showed that the NEVLP group was separated from SCS and HMP groups. Further in-depth analyses indicated significantly (p < 0.05, VIP > 1) higher levels of acylcarnitines, phosphocholines, ether-linked and longer-chain phosphoethanolamines, triacylglycerols and most lysophosphocholines and lysophosphoethanolamines in the hypothermic preservation group. The results showed that the preservation temperature has a more significant impact on the lipidomic profile of the kidney than the preservation method's mechanical characteristics.

Conclusion: Higher levels of lipids detected in the hypothermic preservation group may be related to ischemia-reperfusion injury, mitochondrial dysfunction, pro-inflammatory effect, and oxidative stress. Obtained results suggest the NEVKP method's beneficial effect on graft function and confirm that SPME chemical biopsy enables low-invasive and repeated sampling of the same tissue, allowing tracking alterations in the graft throughout the entire transplantation procedure.

Keywords: LC-MS; SPME; graft quality assessment; kidney perfusion; kidney transplantation; lipidomics; solid-phase microextraction.

Grants and funding

The authors declare that financial support was received for the research, authorship, and/or publication of this article. This study was funded by National Science Centre, grant Opus No. 2017/27/B/NZ5/01013.