Philip M Zakas, Sharon C Cunningham, Ann Doherty, Eva B van Dijk, Raed Ibraheim, Stephanie Yu, Befikadu D Mekonnen, Brendan Lang, Elizabeth J English, Gang Sun, Miles C Duncan, Matthew S Benczkowski, Robert C Altshuler, Malvenderjit Jagjit Singh, Emily S Kibbler, Gulen Y Tonga, Zi Jun Wang, Z Jane Wang, Guangde Li, Ding An, James B Rottman, Yashvi Bhavsar, Cormac Purcell, Rachit Jain, Ryan Alberry, Nathaniel Roquet, Yanfang Fu, Robert J Citorik, Jacob R Rubens, Michael C Holmes, Cecilia Cotta-Ramusino, William Querbes, Ian E Alexander, William E Salomon
{"title":"睡美人 mRNA-LNP 可使 rAAV 转基因在小鼠和 NHP 肝细胞中稳定表达,并提高载体效力。","authors":"Philip M Zakas, Sharon C Cunningham, Ann Doherty, Eva B van Dijk, Raed Ibraheim, Stephanie Yu, Befikadu D Mekonnen, Brendan Lang, Elizabeth J English, Gang Sun, Miles C Duncan, Matthew S Benczkowski, Robert C Altshuler, Malvenderjit Jagjit Singh, Emily S Kibbler, Gulen Y Tonga, Zi Jun Wang, Z Jane Wang, Guangde Li, Ding An, James B Rottman, Yashvi Bhavsar, Cormac Purcell, Rachit Jain, Ryan Alberry, Nathaniel Roquet, Yanfang Fu, Robert J Citorik, Jacob R Rubens, Michael C Holmes, Cecilia Cotta-Ramusino, William Querbes, Ian E Alexander, William E Salomon","doi":"10.1016/j.ymthe.2024.06.021","DOIUrl":null,"url":null,"abstract":"<p><p>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<sup>ash</sup> 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.</p>","PeriodicalId":19020,"journal":{"name":"Molecular Therapy","volume":null,"pages":null},"PeriodicalIF":12.1000,"publicationDate":"2024-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Sleeping Beauty mRNA-LNP enables stable rAAV transgene expression in mouse and NHP hepatocytes and improves vector potency.\",\"authors\":\"Philip M Zakas, Sharon C Cunningham, Ann Doherty, Eva B van Dijk, Raed Ibraheim, Stephanie Yu, Befikadu D Mekonnen, Brendan Lang, Elizabeth J English, Gang Sun, Miles C Duncan, Matthew S Benczkowski, Robert C Altshuler, Malvenderjit Jagjit Singh, Emily S Kibbler, Gulen Y Tonga, Zi Jun Wang, Z Jane Wang, Guangde Li, Ding An, James B Rottman, Yashvi Bhavsar, Cormac Purcell, Rachit Jain, Ryan Alberry, Nathaniel Roquet, Yanfang Fu, Robert J Citorik, Jacob R Rubens, Michael C Holmes, Cecilia Cotta-Ramusino, William Querbes, Ian E Alexander, William E Salomon\",\"doi\":\"10.1016/j.ymthe.2024.06.021\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>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<sup>ash</sup> 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.</p>\",\"PeriodicalId\":19020,\"journal\":{\"name\":\"Molecular Therapy\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":12.1000,\"publicationDate\":\"2024-07-02\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Molecular Therapy\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.1016/j.ymthe.2024.06.021\",\"RegionNum\":1,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"BIOTECHNOLOGY & APPLIED MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Molecular Therapy","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1016/j.ymthe.2024.06.021","RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"BIOTECHNOLOGY & APPLIED MICROBIOLOGY","Score":null,"Total":0}
Sleeping Beauty mRNA-LNP enables stable rAAV transgene expression in mouse and NHP hepatocytes and improves vector potency.
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.
期刊介绍:
Molecular Therapy is the leading journal for research in gene transfer, vector development, stem cell manipulation, and therapeutic interventions. It covers a broad spectrum of topics including genetic and acquired disease correction, vaccine development, pre-clinical validation, safety/efficacy studies, and clinical trials. With a focus on advancing genetics, medicine, and biotechnology, Molecular Therapy publishes peer-reviewed research, reviews, and commentaries to showcase the latest advancements in the field. With an impressive impact factor of 12.4 in 2022, it continues to attract top-tier contributions.