Effect of LEF1 overexpression on gene expression of human Natural Killer T cells

Va24-invariant natural killer T cells (NKTs) possess innate antitumor properties that can be exploited for cancer immunotherapy. We have shown that the CD62L+ central memory-like subset drives NKT in vivo anti-tumor activity, but molecular mediators of NKT central memory differentiation remain unknown. Here, we demonstrate that CD62L+ NKTs express Wnt/ß-catenin transcription factor LEF1 and maintain active Wnt/ß-catenin signaling. CRISPR/Cas9-mediated LEF1 knockout reduced CD62L+ frequency after antigenic stimulation, while Wnt/ß-catenin activator Wnt3a ligand increased CD62L+ frequency. LEF1 overexpression promoted NKT expansion and limited exhaustion following serial tumor challenge and was sufficient to induce a central memory-like transcriptional program in NKTs. In mice, NKTs expressing a GD2-specific chimeric antigen receptor (CAR) with LEF1 demonstrated superior control of neuroblastoma xenograft tumors compared to CAR-NKTs. These results identify LEF1 as a transcriptional activator of the NKT central memory program and advance development of NKT cell-based immunotherapy. Overall design: Comparative gene expression profiling analysis of RNA-seq data for NKTs with LEF1 overexpression versus control luciferase overexpression