RNA sequencing of WT and TrIP KO P14 TCR Tg T cells stimulated in vitro

The protein PI3K-interacting protein (PIK3IP1), or transmembrane inhibitor of PI3K (TrIP), is highly expressed by T cells and can modulate PI3K activity in these cells. Several studies have also revealed that TrIP is rapidly downregulated following T cell activation and can play important roles in T cell differentiation. We have generated mice with CD8-specific TrIP deficiency. Here we provide data detailing that activated TrIP KO CD8 T cells display an increased inflammatory transcriptional profile in the absence of TrIP. Consistent with these effects, we also show that knockout of TrIP specifically in CD8 T cells resulted in reduced growth of syngeneic tumors. When characterizing the tumor-infiltrating cells, we found that TrIP KO led to an increase in the number of tumor-infiltrating T cells, as well as a delay in the acquisition of an exhausted phenotype, based on phenotypic and transcriptomic analyses. Finally, our data suggest that TrIP regulates the diversity of T cell clonal responses to tumors, since we observed an increase in the number of distinct T cell clonotypes responding to a tumor neoantigen. Taken together, we show that TrIP intrinsically restricts the CD8 T cell response to tumors, and that targeting TrIP may augment the anti-tumor response in a way that is distinct from established checkpoint therapies. Overall design: Splenocytes from WT and CD8-specific TrIP knockout P14 mice were cultured for 24 hrs under 3 different conditions; Unstimulated (50 U/ml rIL-2 + 2 ng/ml rIL-12), Stimulated (100 ng/ml HiAff gp33 peptide + 50 U/ml rIL-2 + 2 ng/ml rIL-12), and stimulated + PI3Ki (10 µM IC87114 + 100 ng/ml HiAff gp33 peptide + 50 U/ml rIL-2 + 2 ng/ml rIL-12). After 24 hr stim., CD8 T cells were sorted out and bulk RNAseq was performed.