Normothermic perfusion of human livers for profiling lentiviral vector pharmacokinetics and transduction

Journal article

Lentiviral vectors (LVs) hold significant potential for gene therapy (GT) due to their ability to integrate into non-dividing cells, potentially offering lifelong cures from a single dose. The liver is an attractive GT target due to its role in inherited disorders and as a sink for intravenously delivered therapeutics. However, clinical translation of LV therapy remains challenging due to the poor predictive value of animal models. Normothermic machine perfusion (NMP) maintains human organs under physiological conditions ex vivo, creating an opportunity to reduce reliance on animal studies and de-risk human clinical trials. We used NMP to assess pharmacokinetics and transduction in four human livers dosed with a LV encoding green fluorescent protein (GFP). Perfusion was maintained for up to 74 hours, achieving physiological viability and function. Rapid LV clearance was observed, with less than 1% remaining in perfusate after 10 minutes. Integrated viral copy number per cell (VCN/cell) reached 0.07–0.13, with detectable GFP expression. Transcriptomic analysis revealed dynamic changes in metabolic and inflammatory pathways correlating with liver function and transduction outcomes. This study demonstrates that NMP provides a useful model to assess LV delivery and transduction, supporting its potential as a platform to enhance translation into human clinical applications.

Authors: Nicholls, BRM; Johnson, D; Kulkarni, A; Raposo, RAS; Mitrophanous, KA

• Publication status: Published
• Peer review status: Peer reviewed
• Publisher: Cell Press
• Journal: Molecular Therapy Advances
• Volume: 34
• Issue: 1
• Article number: 201660
• Publication date: 2025-12-26
• Acceptance date: 2025-12-18
• DOI: 10.1016/j.omta.2025.201660
• EISSN: 3117-387X
• Language: English
• Keywords: lentiviral vector; gene therapy; liver-directed gene therapy; normothermic machine perfusion; ex vivo; pharmacokinetics; pharmacodynamics; liver; pre-clinical model