Ex Vivo Machine Perfusion as a Platform for Lentiviral Gene Delivery in Rat Livers

TL;DR

Researchers used machine perfusion to successfully deliver genes into rat livers, opening new possibilities for organ modification before transplantation.

Contribution

This study demonstrates the feasibility of using ex vivo machine perfusion for lentiviral gene delivery in rodent livers.

Findings

Lentiviral vectors successfully delivered genes to rat livers during 72 hours of machine perfusion.

Perfused livers showed significantly increased luminescence, indicating successful genetic modification.

The study validated ex vivo genetic modification as a platform for future organ manipulation.

Abstract

Developing new strategies for local monitoring and delivery of immunosuppression
is critical to making allografts safer and more accessible. Ex vivo genetic modification
of grafts using machine perfusion presents a promising approach to improve graft function
and modulate immune responses while minimizing risks of off-target effects and systemic
immunogenicity in vivo. This proof-of-concept study demonstrates the feasibility of using
normothermic machine perfusion (NMP) to mimic in vitro conditions for effective gene
delivery. In this study, lentiviral vectors carrying biosensor constructs with Gaussia
Luciferase (GLuc) were introduced to rodent livers during a 72-hour perfusion period, with
a targeted delivery of 3 × 107 infection units (IU). Following the
initial 24-hour exposure required for viral transduction, an additional 48 hours was
necessary to observe gene expression, analogous to in vitro benchmarks. The perfused
livers displayed significantly increased luminescence compared to controls, illustrating
successful genetic modification. These findings validate the ex vivo use of lentiviral
particles in a rodent liver model and lay the groundwork for a broad range of applications
through genetic manipulation of organ systems. Future studies will focus on refining this
technology to enhance precision in gene expression and explore its implications for
clinical transplantation.

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