{Reference Type}: Journal Article
{Title}: Sleeping Beauty mRNA-LNP enables stable rAAV transgene expression in mouse and NHP hepatocytes and improves vector potency.
{Author}: 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;Salomon WE;
{Journal}: Mol Ther
{Volume}: 0
{Issue}: 0
{Year}: 2024 Jul 2
{Factor}: 12.91
{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.