Mitochondrial injury during normothermic regional perfusion (NRP) and hypothermic oxygenated perfusion (HOPE) in a rodent model of DCD liver transplantation

Rebecca Panconesi; Mauricio Flores Carvalho; Janina Eden; Marilena Fazi; Fariha Ansari; Leandro Mancina; Nadia Navari; Richard Xavier Sousa Da Silva; Daniele Dondossola; Lucia Bautista Borrego; Matthias Pietzke; Adriano Peris; David Meierhofer; Paolo Muiesan; Alexander Galkin; Fabio Marra; Philipp Dutkowski; Andrea Schlegel

doi:10.1016/j.ebiom.2023.104861

Mitochondrial injury during normothermic regional perfusion (NRP) and hypothermic oxygenated perfusion (HOPE) in a rodent model of DCD liver transplantation

EBioMedicine. 2023 Dec:98:104861. doi: 10.1016/j.ebiom.2023.104861. Epub 2023 Nov 3.

Authors

Rebecca Panconesi¹, Mauricio Flores Carvalho², Janina Eden³, Marilena Fazi⁴, Fariha Ansari⁵, Leandro Mancina³, Nadia Navari², Richard Xavier Sousa Da Silva³, Daniele Dondossola⁶, Lucia Bautista Borrego³, Matthias Pietzke⁷, Adriano Peris⁸, David Meierhofer⁷, Paolo Muiesan⁹, Alexander Galkin⁵, Fabio Marra¹⁰, Philipp Dutkowski³, Andrea Schlegel¹¹

Affiliations

¹ Hepatobiliary Unit, Careggi University Hospital, University of Florence, Florence, Italy; Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy; Department of Surgery, A.O.U. Città della Salute e della Scienza di Torino, University of Turin, 10124, Turin, Italy; Department of Visceral Surgery and Transplantation, University Hospital Zurich, Swiss HPB and Transplant Center, Zurich, Switzerland.
² Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy.
³ Department of Visceral Surgery and Transplantation, University Hospital Zurich, Swiss HPB and Transplant Center, Zurich, Switzerland.
⁴ Hepatobiliary Unit, Careggi University Hospital, University of Florence, Florence, Italy; Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy.
⁵ Feil Family Brain and Mind Research Institute, Weill Cornell Medicine, 407 East 61st Street, New York, NY, 10065, USA.
⁶ General and Liver Transplant Surgery Unit, Fondazione IRCCS Ca' Granda, Ospedale Maggiore Policlinico and University of Milan, Centre of Preclinical Research, 20122, Italy.
⁷ Max Planck Institute for Molecular Genetics, Mass Spectrometry Facility, 14195, Berlin, Germany.
⁸ Tuscany Regional Transplant Authority, Centro Regionale Allocazione Organi e Tessuti (CRAOT), Florence, Italy.
⁹ Hepatobiliary Unit, Careggi University Hospital, University of Florence, Florence, Italy; Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy; Feil Family Brain and Mind Research Institute, Weill Cornell Medicine, 407 East 61st Street, New York, NY, 10065, USA.
¹⁰ Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy; Center for Research, High Education and Transfer DENOThe, University of Florence, Florence, Italy.
¹¹ Hepatobiliary Unit, Careggi University Hospital, University of Florence, Florence, Italy; Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy; Department of Visceral Surgery and Transplantation, University Hospital Zurich, Swiss HPB and Transplant Center, Zurich, Switzerland; General and Liver Transplant Surgery Unit, Fondazione IRCCS Ca' Granda, Ospedale Maggiore Policlinico and University of Milan, Centre of Preclinical Research, 20122, Italy; Transplantation Center, Digestive Disease and Surgery Institute and Department of Immunology, Lerner Research Institute, Cleveland Clinic, Cleveland, OH, USA. Electronic address: schlega4@ccf.org.

Abstract

Background: Normothermic regional perfusion (NRP) and hypothermic-oxygenated-perfusion (HOPE), were both shown to improve outcomes after liver transplantation from donors after circulatory death (DCD). Comparative clinical and mechanistical studies are however lacking.

Methods: A rodent model of NRP and HOPE, both in the donor, was developed. Following asystolic donor warm ischemia time (DWIT), the abdominal compartment was perfused either with a donor-blood-based-perfusate at 37 °C (NRP) or with oxygenated Belzer-MPS at 10 °C (donor-HOPE) for 2 h. Livers were then procured and underwent 5 h static cold storage (CS), followed by transplantation. Un-perfused and HOPE-treated DCD-livers (after CS) and healthy livers (DBD) with direct implantation after NRP served as controls. Endpoints included the entire spectrum of ischemia-reperfusion-injury.

Findings: Healthy control livers (DBD) showed minimal signs of inflammation during 2 h NRP and achieved 100% posttransplant recipient survival. In contrast, DCD livers with 30 and 60 min DWIT suffered from greater mitochondrial injury and inflammation as measured by increased perfusate Lactate, FMN- and HMGB-1-levels with subsequent Toll-like-receptor activation during NRP. In contrast, donor-HOPE (instead of NRP) led to significantly less mitochondrial-complex-I-injury and inflammation. Results after donor-HOPE were comparable to ex-situ HOPE after CS. Most DCD-liver recipients survived when treated with one HOPE-technique (86%), compared to only 40% after NRP (p = 0.0053). Following a reduction of DWIT (15 min), DCD liver recipients achieved comparable survivals with NRP (80%).

Interpretation: High-risk DCD livers benefit more from HOPE-treatment, either immediately in the donor or after cold storage. Comparative prospective clinical studies are required to translate the results.

Funding: Funding was provided by the Swiss National Science Foundation (grant no: 32003B-140776/1, 3200B-153012/1, 320030-189055/1, and 31IC30-166909) and supported by University Careggi (grant no 32003B-140776/1) and the OTT (grant No.: DRGT641/2019, cod.prog. 19CT03) and the Max Planck Society. Work in the A.G. laboratory was partially supported by the NIH R01NS112381 and R21NS125466 grants.

Keywords: Donation after circulatory death; Hypothermic oxygenated perfusion; Ischemia reperfusion injury; Liver transplantation; Mitochondria; Normothermic regional perfusion.

MeSH terms

Animals
Graft Survival
Humans
Inflammation
Liver
Liver Transplantation* / adverse effects
Liver Transplantation* / methods
Organ Preservation / methods
Perfusion / methods
Prospective Studies
Rodentia
Tissue Donors

Abstract

MeSH terms

Grants and funding