Dataset Information

Doxycycline alters porcine renal proteome and degradome during hypothermic machine perfusion

ABSTRACT: Ischemia/reperfusion injury (IRI) is a hallmark of tissue injury in donation after circulatory death (DCD) kidneys. The implementation of hypothermic machine perfusion (HMP) provides a platform for repair of DCD kidneys. Doxycycline administration has shown protective effects during IRI. Here, we explore the impact of doxycycline on proteolytic degradation mechanisms and the urinary proteome of perfused kidney grafts. Porcine kidneys underwent 30 min of warm ischemia, 24 h of oxygenated HMP (control/doxycycline) and 240 min of ex-vivo reperfusion. A high-efficiency undecanal-based N-termini enrichment workflow (HUNTER) was performed to identify the tissue degradome. In addition, a proteomic analysis was applied on urine samples collected during reperfusion using label-free quantitation (LFQ) mass spectrometry. Hierarchical clustering showed distinctive clustering profiles between urine samples collected at T15 min (metabolism) and T240 min (complement and coagulation). At T10, significantly more degraded proteins were observed in perfusates of the control group. At T240, significantly more degraded proteins were observed in the doxycycline group, indicating that doxycycline alters protein degradation during HMP. In conclusion, doxycycline administration during HMP led to significant proteomic differences including protective effects by attenuating urinary NGAL levels. Ultimately, we unravelled multiple pathways that undergo alterations during machine perfusion that can be targeted to attenuate IRI induced injury.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Sus Scrofa Domesticus (domestic Pig)

TISSUE(S): Kidney

SUBMITTER: Leonie van Leeuwen

LAB HEAD: Prof. Dr. B.M. Kessler

PROVIDER: PXD029599 | Pride | 2022-08-12

REPOSITORIES: Pride

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			Action	DRS
	BackgroundLinePos.ami	Other
	BackgroundProfPos.ami	Other
	DensViewPos.ami	Other
	DensViewPosBgnd.ami	Other
	Doxy_proteomics_combined__TMT_labels_only__PeptideGroups.txt	Txt

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Doxycycline Alters the Porcine Renal Proteome and Degradome during Hypothermic Machine Perfusion.

van Leeuwen Leonie L Venema Leonie H LH Heilig Raphael R Leuvenink Henri G D HGD Kessler Benedikt M BM

Current issues in molecular biology 20220123 2

Ischemia-reperfusion injury (IRI) is a hallmark for tissue injury in donation after circulatory death (DCD) kidneys. The implementation of hypothermic machine perfusion (HMP) provides a platform for improved preservation of DCD kidneys. Doxycycline administration has shown protective effects during IRI. Therefore, we explored the impact of doxycycline on proteolytic degradation mechanisms and the urinary proteome of perfused kidney grafts. Porcine kidneys underwent 30 min of warm ischemia, 24 h ...[more]

PMID: 35723325

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