2022 Annual Meeting
Improving Gene Therapy Treatments Via Inhibition of IRF1
Gene therapy advances have paved the way for treatment of many different diseases from cancer to cardiovascular issues. With all of the benefits, gene therapy does have some limitations, including IRF1. IRF1 induces specific protein coding genes that are responsible for inhibiting delivery, transcription, and translation of transgenes. Preliminary experiments with multiple cell lines reveal that in PC-3 cells and primary T-cells, IRF1 transfection efficiency with Lipofectamine and IRF1 expression have an inverse correlation. The hypothesis that drives this work is that transgene expression will increase if IRF1 is inhibited, preventing other host cell genes from expressing. Methods for inhibiting IRF1 include ongoing Cas9 knockouts, small molecule inhibitors, and short hairpin RNAs. A panel of inhibitors have been tested at a range of concentrations to identify the inhibitor(s) that can provide the highest increase in transgene expression with a minimal decrease in cell viability. Inhibitor X has shown preliminary success in 10nM, 100nM, 1uM, and 10uM concentrations by increasing transfection efficiency (i.e., %GFP+ cells) from around 5% without the drug, to around 20% with Inhibitor X added. As an additional approach, shRNA plasmids have been designed and produced to inhibit the translation of IRF1 and preliminary trials have been performed.