1. Sleeping Beauty mRNA-LNP enables stable rAAV transgene expression in mouse and NHP hepatocytes and improves vector potency.
- Author
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Zakas PM, Cunningham SC, Doherty A, van Dijk EB, Ibraheim R, Yu S, Mekonnen BD, Lang B, English EJ, Sun G, Duncan MC, Benczkowski MS, Altshuler RC, Singh MJ, Kibbler ES, Tonga GY, Wang ZJ, Wang ZJ, Li G, An D, Rottman JB, Bhavsar Y, Purcell C, Jain R, Alberry R, Roquet N, Fu Y, Citorik RJ, Rubens JR, Holmes MC, Cotta-Ramusino C, Querbes W, Alexander IE, and Salomon WE
- Subjects
- Animals, Mice, Genetic Therapy methods, Humans, Gene Expression, Lipids chemistry, Disease Models, Animal, Gene Transfer Techniques, Ornithine Carbamoyltransferase genetics, Ornithine Carbamoyltransferase metabolism, Liposomes, Dependovirus genetics, Genetic Vectors genetics, Genetic Vectors administration & dosage, Hepatocytes metabolism, Transgenes, Transposases genetics, Transposases metabolism, Nanoparticles chemistry, RNA, Messenger genetics, RNA, Messenger metabolism
- Abstract
Recombinant adeno-associated virus (rAAV) vector gene delivery systems have demonstrated great promise in clinical trials but continue to face durability and dose-related challenges. Unlike rAAV gene therapy, integrating gene addition approaches can provide curative expression in mitotically active cells and pediatric populations. We explored a novel in vivo delivery approach based on an engineered transposase, Sleeping Beauty (SB100X), delivered as an mRNA within a lipid nanoparticle (LNP), in combination with an rAAV-delivered transposable transgene. This combinatorial approach achieved correction of ornithine transcarbamylase deficiency in the neonatal Spf
ash mouse model following a single delivery to dividing hepatocytes in the newborn liver. Correction remained stable into adulthood, while a conventional rAAV approach resulted in a return to the disease state. In non-human primates, integration by transposition, mediated by this technology, improved gene expression 10-fold over conventional rAAV-mediated gene transfer while requiring 5-fold less vector. Additionally, integration site analysis confirmed a random profile while specifically targeting TA dinucleotides across the genome. Together, these findings demonstrate that transposable elements can improve rAAV-delivered therapies by lowering the vector dose requirement and associated toxicity while expanding target cell types., Competing Interests: Declaration of interests All authors except for S.C.C., E.B.v.D., and I.E.A. are or were employees of Tessera Therapeutics and currently receive or previously received salary along with stock options as compensation for their employment. I.E.A. is a consultant of Tessera Therapeutics, and this work was performed under a Tessera Therapeutics-sponsored research agreement., (Copyright © 2024 The American Society of Gene and Cell Therapy. All rights reserved.)- Published
- 2024
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