High Throughput Proteomic Exploration of Hypothermic Preservation Reveals Active Processes within the Cell Associated with Cold Ischemia Kinetic

Int J Mol Sci. 2021 Feb 27;22(5):2384. doi: 10.3390/ijms22052384.

Abstract

The demand for organs to be transplanted increases pressure on procurement centers, to the detriment of organ quality, increasing complications. New preservation protocols are urgently needed, requiring an in-depth understanding of ischemia-reperfusion mechanisms. We performed a proteomic analysis using LC-MS/MS-TOF data analyzed through R software and Cytoscape's ClueGO application, comparing the proteome of kidney endothelial cells, key cell type, subjected to 3, 6, 12, 19, and 24 h of cold ischemia and 6 h reperfusion. Critical pathways such as energy metabolism, cytoskeleton structure/transport system, and gene transcription/translation were modulated. Important time windows were revealed: a-during the first 3 h, central proteins were upregulated within these pathways; b-the majority of these upregulations were maintained until 12 h cold ischemia time (CIT); c-after that time, the overall decrease in protein expression was observed; d-at reperfusion, proteins expressed in response to cold ischemia were all downregulated. This shows that cold ischemia is not a simple slowing down of metabolism, as deep changes take place within the proteome on major pathways. Time-sensitive expression of key protein reveals possible quality biomarkers as well as potential targets for new strategies to maintain or optimize organ quality.

Keywords: cellular mechanism; ischemia-reperfusion; metabolism; organ preservation; proteomics.

MeSH terms

  • Chromatography, Liquid
  • Cold Ischemia / adverse effects*
  • Cryopreservation / methods*
  • Endothelial Cells / metabolism*
  • Endothelial Cells / pathology
  • High-Throughput Screening Assays
  • Humans
  • Kidney / metabolism*
  • Kidney / pathology
  • Organ Preservation Solutions
  • Proteome / analysis
  • Proteome / metabolism*
  • Reperfusion Injury / etiology
  • Reperfusion Injury / metabolism*
  • Reperfusion Injury / pathology
  • Tandem Mass Spectrometry

Substances

  • Organ Preservation Solutions
  • Proteome