Journal Article

Therapeutic homology-independent targeted integration in retina and liver

Llado-Santaeularia, Manel, Cacchiarelli, Davide, Nigro, Vincenzo, Manfredi, Anna, Tornabene, Patrizia, Iuliano, Antonella, Torella, Annalaura, Rossi, Settimio, Musacchia, Francesco, Dell’Anno, Margherita, Centrulo, Miriam, Nusco, Edoardo, Di Filippo, Lucio, Iodice, Carolina, Ferla, Rita, Surace, Enrico Maria, Auricchio, Alberto, Pone, Emanuela, Marrocco, Elena, Minopoli, Renato, Lyubenova, Hristiana, Esposito, Federica, Piluso, Giulio

2022

Overview

Challenges to the widespread application of gene therapy with adeno-associated viral (AAV) vectors include dominant conditions due to gain-of-function mutations which require allele-specific knockout, as well as long-term transgene expression from proliferating tissues, which is hampered by AAV DNA episomal status. To overcome these challenges, we used CRISPR/Cas9-mediated homology-independent targeted integration (HITI) in retina and liver as paradigmatic target tissues. We show that AAV-HITI targets photoreceptors of both mouse and pig retina, and this results in significant improvements to retinal morphology and function in mice with autosomal dominant retinitis pigmentosa. In addition, we show that neonatal systemic AAV-HITI delivery achieves stable liver transgene expression and phenotypic improvement in a mouse model of a severe lysosomal storage disease. We also show that HITI applications predominantly result in on-target editing. These results lay the groundwork for the application of AAV-HITI for the treatment of diseases affecting various organs. Limits of AAV-mediated gene therapy include targeting dominant mutations and inducing long-term transgene expression. Here, the authors show that AAV-HITI results in efficient allele-independent integration of a donor DNA in both retina and liver providing therapeutic benefit in mouse models of either a genetic form of blindness or a lysosomal storage disease, respectively.

• Publisher: Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
• Subject: 13/31 / 14/63 / 38 / 38/1 / 42/44 / 45 / 45/23 / 45/29 / 45/77 / 49 / 64/60 / 692/698/1688/512/2613/1786 / 692/699/317 / 692/700/565/201/2110 / 82/80 / 9/30 / Alleles / Animal models / Animals / Blindness / CRISPR / CRISPR-Cas Systems / Deoxyribonucleic acid / Dependovirus - genetics / DNA / Expression vectors / Gene Editing - methods / Gene therapy / Genetic Vectors - genetics / Homology / Humanities and Social Sciences / Integration / Liver / Lysosomal storage diseases / Medical treatment / Mice / multidisciplinary / Mutation / Neonates / Organs / Photoreceptors / Retina / Retina - metabolism / Retinitis / Retinitis pigmentosa / Science / Science (multidisciplinary) / Swine