Chimeric Antigen Receptor (CAR) T cells achieved remarkable success in treating B-cell malignancies, yet, their long-term efficiency does not hold up to the original promises. Novel designs of CAR-T cells are urgently needed with further genetic editing for enhanced activity and persistence.
We will elucidate the precise function of RASAL3, a negative regulator of T-cell signaling, and will assess its potential for augmenting CAR-T cell activity. We will generate RASAL3 knockout CAR-T cells to evaluate their signaling, activation strength and killing potential against malignant B cell lines in vitro and their activity, persistence and long-term effect in a mouse model. In addition, using a CRISPR/Cas9 knockout library, we will interrogate genes on a genome-wide level that are able to modulate and fully unleash the activity of CAR-T cells. Identified genes will be thoroughly validated and the underlying molecular mechanisms elucidated. They will be used to develop innovative designs of CAR-T cells, whose performance will be evaluated both in vitro and in vivo.