T cell responses to hub gene knockout in in-vitro propagated Jurkat Cells: Application to Systems Biology (141.50)

Abstract Systems biology is unraveling complex gene interconnections. T cell genes are upregulated during rejection. Suppression of hub and node genes in the T cell genome could result in T cell hyporesponsiveness. Objective: To knockdown hub or node genes via RNA interference in Jurkat cells and study their effect on T cell proliferation. Methods: A molecular interaction network was built with Cytoscape using microarray data on Jurkat cells to identify upregulated hub genes: ATP5C1, IL6ST, PRKCZ, MYC, and MAPK1 that served as targets for RNAi knockdown. Jurkat cell lines were lipotransfected with siRNAs for the selected hub genes. Cell proliferation was measured by ATP release bioluminescence assay and BRDU labeling. Results: At 12 hours following incubation with siRNA MAPK1, there was a reduction in ATP production compared to controls. At 24 hours, siRNAs incorporating genes for ATP5C1, IL6ST, PRKCZ, and MYC showed reduction in Jurkat cell proliferation compared to controls. No difference was observed in MAPK1 group and in the group transfected with a combination of siRNAs for all the identified hub genes. Conclusions: Downregulation of hub genes by siRNA transfection is possible and could open opportunities to target these genes and modify T cell responses. This will help us in integrating systems biology to identify genes that could be potential therapeutic targets.