Bioengineering novel AAV9-mGULO-GT for multi-disease gene therapy: Targeting mutated GULO expression to cure scurvy and brain diseases.

Abstract Current clinical breakthroughs in gene therapy have brought adeno-associated virus (AAV) vectors to the forefront of gene delivery systems. Vitamin C deficiency due to GULO mutations is a genetic disorder affecting guinea pigs and humans. In our study, we used AAV9-mGULO-GT to deliver the mouse GULO gene to guinea pigs and restore Vc synthesis in affected tissues, including the liver and brain. AAV9-mGULO-GT treatment significantly improved survival rates and bone health compared to non-treated and Vc-treated groups. Dot blot analysis confirmed restored Vc content in various parts of the brain. Additionally, micro-CT imaging showcased significant enhancements in bone mineral density, content, width, and cortical thickness. Further, RNA sequencing and immunological studies of organs validated the successful restoration of Vc synthesis. These findings highlight the potential of AAV9-mGULO-GT as a therapeutic option for GULO-related scurvy and other genetic disorders. The success of our study underscores the importance of advanced targeting and gene rescue systems in developing effective therapies for genetic disorders in clinical applications.