Mitochondrial respiration during normothermic liver machine perfusion predicts clinical outcome

Andras T Meszaros; Julia Hofmann; Madita L Buch; Benno Cardini; Theresia Dunzendorfer-Matt; Florian Nardin; Michael J Blumer; Margot Fodor; Martin Hermann; Bettina Zelger; Giorgi Otarashvili; Melanie Schartner; Annemarie Weissenbacher; Rupert Oberhuber; Thomas Resch; Jakob Troppmair; Dietmar Fner; Heinz Zoller; Herbert Tilg; Erich Gnaiger; Theresa Hautz; Stefan Schneeberger

doi:10.1016/j.ebiom.2022.104311

Mitochondrial respiration during normothermic liver machine perfusion predicts clinical outcome

EBioMedicine. 2022 Nov:85:104311. doi: 10.1016/j.ebiom.2022.104311. Epub 2022 Oct 29.

Authors

Andras T Meszaros¹, Julia Hofmann¹, Madita L Buch¹, Benno Cardini¹, Theresia Dunzendorfer-Matt², Florian Nardin³, Michael J Blumer⁴, Margot Fodor¹, Martin Hermann¹, Bettina Zelger⁵, Giorgi Otarashvili¹, Melanie Schartner¹, Annemarie Weissenbacher¹, Rupert Oberhuber¹, Thomas Resch¹, Jakob Troppmair¹, Dietmar Fner¹, Heinz Zoller⁶, Herbert Tilg⁶, Erich Gnaiger⁷, Theresa Hautz¹, Stefan Schneeberger⁸

Affiliations

¹ Department of Visceral, Transplant and Thoracic Surgery, organLife... Laboratory and Daniel Swarovski Research Laboratory, Center of Operative Medicine, Medical University of Innsbruck, Innsbruck, Austria.
² Institute of Biological Chemistry, Biocenter, Medical University of Innsbruck, Innsbruck, Austria.
³ Department of Visceral, Transplant and Thoracic Surgery, organLife... Laboratory and Daniel Swarovski Research Laboratory, Center of Operative Medicine, Medical University of Innsbruck, Innsbruck, Austria; Department of Anatomy, Histology and Embryology, Division of Clinical and Functional Anatomy, Medical University of Innsbruck, Innsbruck, Austria.
⁴ Department of Anatomy, Histology and Embryology, Division of Clinical and Functional Anatomy, Medical University of Innsbruck, Innsbruck, Austria.
⁵ Institute of Pathology, Neuropathology and Molecular Pathology, Medical University of Innsbruck, Innsbruck, Austria.
⁶ Department of Internal Medicine I, Medical University of Innsbruck, Innsbruck, Austria.
⁷ Oroboros Instruments, Innsbruck, Austria.
⁸ Department of Visceral, Transplant and Thoracic Surgery, organLife... Laboratory and Daniel Swarovski Research Laboratory, Center of Operative Medicine, Medical University of Innsbruck, Innsbruck, Austria. Electronic address: stefan.schneeberger@i-med.ac.at.

Abstract

Background: Reliable biomarkers for organ quality assessment during normothermic machine perfusion (NMP) are desired. ATP (adenosine triphosphate) production by oxidative phosphorylation plays a crucial role in the bioenergetic homeostasis of the liver. Thus, detailed analysis of the aerobic mitochondrial performance may serve as predictive tool towards the outcome after liver transplantation.

Methods: In a prospective clinical trial, 50 livers were subjected to NMP (OrganOx Metra) for up to 24.ßh. Biopsy and perfusate samples were collected at the end of cold storage, at 1.ßh, 6.ßh, end of NMP, and 1.ßh after reperfusion. Mitochondrial function and integrity were characterized by high-resolution respirometry (HRR), AMP, ADP, ATP and glutamate dehydrogenase analysis and correlated with the clinical outcome (L-GrAFT score). Real-time confocal microscopy was performed to assess tissue viability. Structural damage was investigated by histology, immunohistochemistry and transmission electron microscopy.

Findings: A considerable variability in tissue viability and mitochondrial respiration between individual livers at the end of cold storage was observed. During NMP, mitochondrial respiration with succinate and tissue viability remained stable. In the multivariate analysis of the 35 transplanted livers (15 were discarded), area under the curve (AUC) of LEAK respiration, cytochrome c control efficiency (mitochondrial outer membrane damage), and efficacy of the mitochondrial ATP production during the first 6.ßh of NMP correlated with L-GrAFT.

Interpretations: Bioenergetic competence during NMP plays a pivotal role in addition to tissue injury markers. The AUC for markers of outer mitochondrial membrane damage, ATP synthesis efficiency and dissipative respiration (LEAK) predict the clinical outcome upon liver transplantation.

Funding: This study was funded by a Grant from the In Memoriam Dr. Gabriel Salzner Stiftung awarded to SS and the Tiroler Wissenschaftsfond granted to TH.

Keywords: High-resolution respirometry; Liver; Mitochondria; Normothermic machine perfusion; Transplantation.

Publication types

Clinical Trial

MeSH terms

Adenosine Triphosphate / metabolism
Cold Ischemia*
Humans
Liver / metabolism
Mitochondria
Organ Preservation*
Perfusion
Prospective Studies
Respiration

Substances

Adenosine Triphosphate