Hypothermic oxygenated perfusion protects from mitochondrial injury before liver transplantation.
Animals
Biomarkers
Cellular Reprogramming
Electron Transport Chain Complex Proteins
/ chemistry
Energy Metabolism
Humans
Hypothermia, Induced
/ methods
Liver
/ metabolism
Liver Function Tests
Liver Transplantation
/ methods
Mitochondria
/ metabolism
Models, Animal
NAD
NADP
Oxygen
/ metabolism
Perfusion
/ methods
Rats
Reperfusion Injury
/ prevention & control
Structure-Activity Relationship
Temperature
Transplantation Conditioning
/ methods
Complex I
FMN
Hypothermic oxygenated perfusion
Liver transplantation
Normothermic oxygenated perfusion
Journal
EBioMedicine
ISSN: 2352-3964
Titre abrégé: EBioMedicine
Pays: Netherlands
ID NLM: 101647039
Informations de publication
Date de publication:
Oct 2020
Oct 2020
Historique:
received:
06
06
2020
revised:
06
09
2020
accepted:
07
09
2020
pubmed:
27
9
2020
medline:
20
7
2021
entrez:
26
9
2020
Statut:
ppublish
Résumé
Mitochondrial succinate accumulation has been suggested as key event for ischemia reperfusion injury in mice. No specific data are however available on behavior of liver mitochondria during ex situ machine perfusion in clinical transplant models. We investigated mitochondrial metabolism of isolated perfused rat livers before transplantation. Livers were exposed to warm and cold ischemia to simulate donation after circulatory death (DCD) and organ transport. Subsequently, livers were perfused with oxygenated Belzer-MPS for 1h, at hypothermic or normothermic conditions. Various experiments were performed with supplemented succinate and/or mitochondrial inhibitors. The perfusate, liver tissues, and isolated mitochondria were analyzed by mass-spectroscopy and fluorimetry. Additionally, rat DCD livers were transplanted after 1h hypothermic or normothermic oxygenated perfusion. In parallel, perfusate samples were analysed during HOPE-treatment of human DCD livers before transplantation. Succinate exposure during rat liver perfusion triggered a dose-dependent release of mitochondrial Flavin-Mononucleotide (FMN) and NADH in perfusates under normothermic conditions. In contrast, perfusate FMN was 3-8 fold lower under hypothermic conditions, suggesting less mitochondrial injury during cold re-oxygenation compared to normothermic conditions. HOPE-treatment induced a mitochondrial reprogramming with uploading of the nucleotide pool and effective succinate metabolism. This resulted in a clear superiority after liver transplantation compared to normothermic perfusion. Finally, the degree of mitochondrial injury during HOPE of human DCD livers, quantified by perfusate FMN and NADH, was predictive for liver function. Mitochondrial injury determines outcome of transplanted rodent and human livers. Hypothermic oxygenated perfusion improves mitochondrial function, and allows viability assessment of liver grafts before implantation. detailed information can be found in Acknowledgments.
Sections du résumé
BACKGROUND
BACKGROUND
Mitochondrial succinate accumulation has been suggested as key event for ischemia reperfusion injury in mice. No specific data are however available on behavior of liver mitochondria during ex situ machine perfusion in clinical transplant models.
METHODS
METHODS
We investigated mitochondrial metabolism of isolated perfused rat livers before transplantation. Livers were exposed to warm and cold ischemia to simulate donation after circulatory death (DCD) and organ transport. Subsequently, livers were perfused with oxygenated Belzer-MPS for 1h, at hypothermic or normothermic conditions. Various experiments were performed with supplemented succinate and/or mitochondrial inhibitors. The perfusate, liver tissues, and isolated mitochondria were analyzed by mass-spectroscopy and fluorimetry. Additionally, rat DCD livers were transplanted after 1h hypothermic or normothermic oxygenated perfusion. In parallel, perfusate samples were analysed during HOPE-treatment of human DCD livers before transplantation.
FINDINGS
RESULTS
Succinate exposure during rat liver perfusion triggered a dose-dependent release of mitochondrial Flavin-Mononucleotide (FMN) and NADH in perfusates under normothermic conditions. In contrast, perfusate FMN was 3-8 fold lower under hypothermic conditions, suggesting less mitochondrial injury during cold re-oxygenation compared to normothermic conditions. HOPE-treatment induced a mitochondrial reprogramming with uploading of the nucleotide pool and effective succinate metabolism. This resulted in a clear superiority after liver transplantation compared to normothermic perfusion. Finally, the degree of mitochondrial injury during HOPE of human DCD livers, quantified by perfusate FMN and NADH, was predictive for liver function.
INTERPRETATION
CONCLUSIONS
Mitochondrial injury determines outcome of transplanted rodent and human livers. Hypothermic oxygenated perfusion improves mitochondrial function, and allows viability assessment of liver grafts before implantation.
FUNDING
BACKGROUND
detailed information can be found in Acknowledgments.
Identifiants
pubmed: 32979838
pii: S2352-3964(20)30390-X
doi: 10.1016/j.ebiom.2020.103014
pmc: PMC7519249
pii:
doi:
Substances chimiques
Biomarkers
0
Electron Transport Chain Complex Proteins
0
NAD
0U46U6E8UK
NADP
53-59-8
Oxygen
S88TT14065
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
103014Subventions
Organisme : NINDS NIH HHS
ID : R01 NS112381
Pays : United States
Informations de copyright
Copyright © 2020 The Author(s). Published by Elsevier B.V. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of Interests This study was conducted at the University Hospital Zurich (USZ). Raw data and laboratory analysis results were extracted at the laboratory of the Department of Visceral Surgery and Transplantation of the USZ. Data analysis was carried out at the laboratory of the Department of Visceral Surgery and Transplantation of the USZ. The authors have declared that no competing interests exist. The study was principally funded by a main research grant from the swiss national foundation (SNF), awarded to Professor Philipp Dutkowski (Ref: 32003B-140776/1, 3200B-153012/1, 31IC30-166909). Mitochondrial and tissue analysis was further supported by the Max Planck Society (Dr. David Meierhofer) and the NIH grant: R01NS112381 awarded to Dr. Alexander Galkin. There are no patents involved or affecting this study.
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