Abstract:
Gene therapy has emerged in the last decade as a widely studied and implemented group of unique therapeutic modalities. Though still young and investigational in nature, it has generated significant traction, with many translational technologies making their way to the clinic as licensed treatments for genetic diseases. For in vivo gene therapies, adeno-associated virus (AAV) is among the most widely studied and promising viral vectors, due to its low immunogenicity and diverse tissue tropism. AAV transduction and hence the efficiency of genetic transfer is influenced by host trafficking and processing machinery, through proteins defined as host factors. This study aims at understanding the role played by one such host factor, the WD-repeat family protein WDR11 in AAV transduction. WDR11 was knocked out in different cell lines using CRISPR/Cas9 and these knockout cells were studied for differences in AAV transduction. Genomic knockout of WDR11 increased AAV transduction by 1.5 fold in HeLa cells, and 3-3.5 fold in eHAP cells, an effect that was reversed by ectopic overexpression of WDR11 in knockout cells. This study also showed that WDR11 KO increases the cell surface expression of AAVR, a receptor shown to be critical for transduction by multiple AAV serotypes. It is hence possible that WDR11 KO causes leads to increased localisation of AAVR at the cell surface, and together assisted by compensatory pathways leads to increased AAV trafficking in the cell. Overall, this study aims to study WDR11 as a potential restriction factor to gain a deeper understanding of AAV biology.
