Characterization of the Humanized FRG Mouse Model and Development of an AAV-LK03 Variant with Improved Liver Lobular Biodistribution

Overview

Journal Mol Ther Methods Clin Dev

Publisher Cell Press

Date 2023 Jan 26

PMID 36700121

Authors

Marti Cabanes-Creus

Renina Gale Navarro

Sophia H Y Liao

Suzanne Scott

Rodrigo Carlessi

Ramon Roca-Pinilla

Maddison Knight

Grober Baltazar

Erhua Zhu

Matthew Jones

Elena Denisenko

Alistair R R Forrest

Ian E Alexander

Janina E E Tirnitz-Parker

Leszek Lisowski

Affiliations

Soon will be listed here.

Abstract

Recent clinical successes have intensified interest in using adeno-associated virus (AAV) vectors for therapeutic gene delivery. The liver is a key clinical target, given its critical physiological functions and involvement in a wide range of genetic diseases. In the present study, we first investigated the validity of a liver xenograft mouse model repopulated with primary hepatocytes using single-nucleus RNA sequencing (sn-RNA-seq) by studying the transcriptomic profile of human hepatocytes pre- and post-engraftment. Complementary immunofluorescence analyses performed in highly engrafted animals confirmed that the human hepatocytes organize and present appropriate patterns of zone-dependent enzyme expression in this model. Next, we tested a set of rationally designed HSPG de-targeted AAV-LK03 variants for relative transduction performance in human hepatocytes. We used immunofluorescence, next-generation sequencing, and single-nucleus transcriptomics data from highly engrafted FRG mice to demonstrate that the optimally HSPG de-targeted AAV-LK03 displayed a significantly improved lobular transduction profile in this model.

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