Systemic delivery of AAV5, AAV8, and AAV9 packaging a C5-12-microdystrophin-FLAG expression cassette in non-human primates

Systemic delivery of AAV5, AAV8, and AAV9 packaging a C5-12-microdystrophin-FLAG expression cassette in non-human primates

Safely achieving therapeutic expression levels with adeno-associated virus (AAV) gene therapy is a significant challenge for treating the large muscle mass in humans. Non-human primates (NHPs) provide a more accurate assessment of the feasibility of achieving an effective and safe dose than rodents....

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Main Authors: Mengping Liu, Erica Cook, Yanshan Dai, Erich Ehlert, Francois du Plessis, Jacek Lubelski, Bogdan G. Sleczka, Petia Shipkova, Zhuyin Li, Joshua Gamse, David Gordon, Leonard P. Adam, Paul C. Levesque, Glen B. Banks
Format: Article
Language:English
Published: Elsevier 2025-03-01
Series:Molecular Therapy: Methods & Clinical Development
Subjects:
adeno-associated virus
promoter
systemic delivery
non-human primate
dystrophin
AAV
Online Access:http://www.sciencedirect.com/science/article/pii/S2329050125000063
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Summary:Safely achieving therapeutic expression levels with adeno-associated virus (AAV) gene therapy is a significant challenge for treating the large muscle mass in humans. Non-human primates (NHPs) provide a more accurate assessment of the feasibility of achieving an effective and safe dose than rodents. Here, we compared a single systemic administration of AAV5, AAV8, or AAV9 in NHPs, each packaging the C5-12-microdystrophin-FLAG expression cassette. At 1 month post-dose, we compared tissue vector genomes, mRNA, and microdystrophin-FLAG protein levels by meso-scale discovery-enzyme-linked immunosorbent assay, liquid chromatography-mass spectrometry, and immunofluorescence. The C5-12 promoter was highly selective for heart and skeletal muscles, when compared to off-target tissues such as peripheral blood mononuclear cells, lung, liver, and kidney. AAV8 led to higher levels of microdystrophin-FLAG mRNA and protein in the cardiac ventricles and skeletal muscles when compared to AAV5 or AAV9. The AAV8-microdystrophin-FLAG led to ∼20% of wild-type NHP dystrophin protein expression levels and was located on the sarcolemma of ∼40% of skeletal muscles fibers and ∼15% of left ventricular cardiomyocytes. Hematology, serum chemistry, and pathology were unremarkable. Thus, a systemic dose of ∼1.18 × 1014 vector genomes/kg AAV8 is predicted to be safe and efficacious for treating Duchenne muscular dystrophy (DMD) but has significant room for improvement.
ISSN:2329-0501

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