Sleeping Beauty mRNA-LNP enables stable rAAV transgene expression in mouse and NHP hepatocytes and improves vector potency

  • Mol Ther. 2024 Jul 2:S1525-0016(24)00403-9. doi: 10.1016/j.ymthe.2024.06.021.
Philip M Zakas  1 ,  Sharon C Cunningham  2 ,  Ann Doherty  1 ,  Eva B van Dijk  2 ,  Raed Ibraheim  1 ,  Stephanie Yu  1 ,  Befikadu D Mekonnen  1 ,  Brendan Lang  1 ,  Elizabeth J English  1 ,  Gang Sun  1 ,  Miles C Duncan  1 ,  Matthew S Benczkowski  1 ,  Robert C Altshuler  1 ,  Malvenderjit Jagjit Singh  1 ,  Emily S Kibbler  1 ,  Gulen Y Tonga  1 ,  Zi Jun Wang  1 ,  Z Jane Wang  1 ,  Guangde Li  1 ,  Ding An  1 ,  James B Rottman  1 ,  Yashvi Bhavsar  1 ,  Cormac Purcell  1 ,  Rachit Jain  1 ,  Ryan Alberry  1 ,  Nathaniel Roquet  1 ,  Yanfang Fu  1 ,  Robert J Citorik  1 ,  Jacob R Rubens  1 ,  Michael C Holmes  1 ,  Cecilia Cotta-Ramusino  1 ,  William Querbes  1 ,  Ian E Alexander  3 ,  William E Salomon  4
Affiliations
  • 1. Tessera Therapeutics, Inc., Somerville, MA 02143, USA.
  • 2. Gene Therapy Research Unit, Children's Medical Research Institute, Faculty of Medicine and Health, The University of Sydney and Sydney Children's Hospitals Network, Westmead, NSW 2145, Australia.
  • 3. Gene Therapy Research Unit, Children's Medical Research Institute, Faculty of Medicine and Health, The University of Sydney and Sydney Children's Hospitals Network, Westmead, NSW 2145, Australia; Discipline of Child and Adolescent Health, University of Sydney, Westmead, NSW 2145, Australia. Electronic address: [email protected].
  • 4. Tessera Therapeutics, Inc., Somerville, MA 02143, USA. Electronic address: [email protected].
PMID: 38981468 DOI: 10.1016/j.ymthe.2024.06.021
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 Spfash 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.

Keywords
Sleeping Beauty transposase; adeno-associated virus vector; gene addition; lipid nanoparticle; mRNA liver delivery; macaque; ornithine transcarbamylase; partial hepatectomy; transposition.
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