Characterization of the humanized FRG mouse model and development of an AAV-LK03 variant with improved liver lobular biodistribution

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 investig...

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Vydané v:Molecular therapy. Methods & clinical development Ročník 28; s. 220 - 237
Hlavní autori: Cabanes-Creus, Marti, Navarro, Renina Gale, Liao, Sophia H.Y., Scott, Suzanne, Carlessi, Rodrigo, Roca-Pinilla, Ramon, Knight, Maddison, Baltazar, Grober, Zhu, Erhua, Jones, Matthew, Denisenko, Elena, Forrest, Alistair R.R., Alexander, Ian E., Tirnitz-Parker, Janina E.E., Lisowski, Leszek
Médium: Journal Article
Jazyk:English
Vydavateľské údaje: United States Elsevier Inc 09.03.2023
American Society of Gene & Cell Therapy
Elsevier
Predmet:
adeno-associated vector
gene therapy
humanized FRG
liver lobule
liver zonation
lobular transduction
Original
single-nucleus RNA sequencing
liver lobule
liver zonation
single-nucleus RNA sequencing
gene therapy
lobular transduction
humanized FRG
adeno-associated vector
ISSN:2329-0501, 2329-0501
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Shrnutí: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. [Display omitted] Using single-nucleus RNA sequencing and complementary immunofluorescence analyses, the authors confirmed that human hepatocytes engrafted in xenograft mice organize and present appropriate patterns of liver-zone-dependent enzyme expression. Having confirmed this, the authors successfully performed targeted bioengineering of a clinical AAV variant, AAV-LK03, to improve its lobular transduction profile.
Bibliografia:ObjectType-Article-1
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ISSN:2329-0501
2329-0501
DOI:10.1016/j.omtm.2022.12.014