Inducing Hepatitis C Virus Resistance After Pig Liver Transplantation—A Proof of Concept of Liver Graft Modification Using Warm Ex Vivo Perfusion

Abstract

Normothermic ex vivo liver perfusion (NEVLP) offers the potential to optimize graft function prior to liver transplantation (LT). Hepatitis C virus (HCV) is dependent on the presence of miRNA(microRNA)-122. Miravirsen, a locked-nucleic acid oligonucleotide, sequesters miR-122 and inhibits HCV replication. The aim of this study was to assess the efficacy of delivering miravirsen during NEVLP to inhibit miR-122 function in a pig LT model. Pig livers were treated with miravirsen during NEVLP or cold storage (CS). Miravirsen absorption, miR-122 sequestration, and miR-122 target gene derepression were determined before and after LT. The effect of miravirsen treatment on HCV infection of hepatoma cells was also assessed. NEVLP improved miravirsen uptake versus CS. Significant miR-122 sequestration and miR-122 target gene derepression were seen with NEVLP but not with CS. In vitro data confirmed miravirsen suppression of HCV replication after established infection and prevented HCV infection with pretreatment of cells, analogous to the pretreatment of grafts in the transplant setting. In conclusion, miravirsen delivery during NEVLP is a potential strategy to prevent HCV reinfection after LT. This is the first large-animal study to provide "proof of concept" for using NEVLP to modify and optimize liver grafts for transplantation.

Improvement of Normothermic Ex Vivo Machine Perfusion of Rat Liver Grafts by Dialysis and Kupffer Cell Inhibition With Glycine

“…Normothermic conditions offer the chance for pharmacologic interventions that are dependent on an active metabolism of the graft . For example, Goldaracena et al demonstrated the feasibility of anti‐inflammatory treatment and inhibition of hepatitis C virus replication during NEVLP of porcine liver grafts . However, large animal models of NEVLP are cost intensive and comprise logistic implications.…”

“…(11) For example, Goldaracena et al demonstrated the feasibility of anti-inflammatory treatment and inhibition of hepatitis C virus replication during NEVLP of porcine liver grafts. (23,25) However, large animal models of NEVLP are cost intensive and comprise logistic implications. Standardized small animal models for basic research on machine perfusion of the liver and large sets of dose-response and timing studies on pharmacological conditioning are not available in the literature.…”

Bioengineering approaches to organ preservation ex vivo

“…For example, bioengineering organs through decellularization and repopulation with specific cell types have been explored in ex vivo MP and bioreactor platforms by our group and others, 58,111,[116][117][118] though the current timeline of ex vivo MP does not allow us to effectively assess cell engraftment and function. Extended preservation time could enable development of improved cell-based therapies and allow for cell uptake and engraftment prior to transplantation.…”

Control of Ischemia-Reperfusion Injury in Liver Transplantation: Potentials for Increasing the Donor Pool

“…The main potential of MP is not only simple resuscitation of donor organs but also the possibility to administer organ-specific therapies with the avoidance of systemic side effects, which is in contrast to all previously described direct donor and recipient preconditioning methods. In LT, efforts were reported in experimental settings to inactivate miRNA-122 utilizing normothermic MP (NMP) in order to prevent hepatitis C virus (HCV) infection after transplantation of HCV-positive organs [37], whereas these are of minor importance since the advent of the new, highly effective anti-HCV treatment options.…”