RELB Reprograms Exhausted Tumor-Infiltrating Lymphocytes for Improved Adoptive Cell Therapy

Abstract Tumor-infiltrating lymphocytes (TILs) are a promising autologous cell therapy to treat solid tumors. TILs are manufactured by expanding and reinfusing tumor-reactive T cells from tumor biopsies. Efficacy of TIL therapies has been limited by the heterogeneity of expanded TIL products and the high prevalence of dysfunctional exhausted CD8+ T cells (TEX). While a subset of CD8+ TILs co-expressing CD103 and CD39 are enriched for tumor-reactive TILs across multiple cancer types, these cells are often in the TEX state with low proliferative potential. To identify regulators of human TIL proliferation, we screened an open reading frame library encoding for all human transcription factors (TFs). RELB emerged as the dominant driver of human TIL expansion with a skew towards CD8+ cells. TCR diversity was maintained after multiple days of in vitro expansion driven by RELB. Transcriptome profiling of multiple RELB-expressing TIL subtypes revealed a shift towards a memory/costimulatory-like phenotype. Using a HER2-targeting CAR and tumor co-culture model, RELB conferred improved persistence after multiple tumor challenges in vitro and improved solid tumor control in mouse xenografts in vivo. Finally, co-culture of RELB-overexpressing TILs with patient-matched tumor organoids showed an increase in TIL product polyfunctionality, tumor reactivity, and tumor killing. Collectively these results support promoting RELB expression as a strategy for broadly enabling TIL therapy for treating solid tumors. Competing Interest Statement C.A.G. is a co-founder of Tune Therapeutics, Sollus Therapeutics, and Locus Biosciences and is an advisor to Tune Therapeutics, Sarepta Therapeutics, and Pappas Capital, LLC. S.A. is an advisor to Immutep and a co-founder of Cellepus Therapeutics. C.D.M.A., S.R.J., and C.A.G. are inventors on patents or patent applications related to CRISPR editing technologies, methods for high-throughput screening, and engineering of T cell therapies.