
==== Front
Res Sq
ResearchSquare
Research Square
2693-5015
American Journal Experts

10.21203/rs.3.rs-4784505/v1
10.21203/rs.3.rs-4784505
preprint
1
Article
Ex Vivo Machine Perfusion as a Platform for Lentiviral Gene Delivery in Rat Livers
Uygun Korkut
von Reiterdank Irina Filz https://orcid.org/0000-0002-0234-2704

Mojoudi Mohammadreza
Bento Raphaela
Taggart McLean
Dinicu Antonia
Wojtkiewicz Gregory
Coert J.
van der Molen Aebele Mink
Weissleder Ralph https://orcid.org/0000-0003-0828-4143

Parekkadan Biju
13 9 2024
rs.3.rs-4784505https://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License, which allows reusers to distribute, remix, adapt, and build upon the material in any medium or format, so long as attribution is given to the creator. The license allows for commercial use.
https://www.researchsquare.com/article/rs-4784505/v1
nihpp-rs4784505v1.pdf
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 x 10 7 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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