Peter. P. Challita

Ewing sarcoma (EwS) is an aggressive malignancy of children, adolescents, and young adults driven by the EWS::FLI1 fusion oncoprotein with few effective treatment options in the relapsed or refractory setting. We validate six transmembrane epithelial antigen of the prostate 1 (STEAP1) as a fusion-regulated and clinically actionable surface antigen with broad expression across a series of primary EwS tumors. Analysis of EWS::FLI1 perturbation in EwS models demonstrated regulation of STEAP1 transcript and protein expression by oncogenic EWS::FLI1. Second-generation STEAP1 CAR T cells induced complete tumor regression in orthotopic models and exhibited dose-dependent activity in disseminated disease without evidence of antigen escape. Armoring STEAP1 CAR T cells with IL-18 further potentiated antitumor efficacy, achieving complete responses without overt toxicity in approximately 80% of mice at a five-fold lower cell dose where unarmored STEAP1 CAR T cells were ineffective. Notably, adoptive transfer of autologous STEAP1 CAR T cells is currently being evaluated in a first-in-human phase I/II clinical trial in metastatic castration-resistant prostate cancer (NCT06236139) and the compelling preclinical data presented here support clinical translation to relapsed/refractory EwS.

Tissue-resident memory CD8 T (T_RM) cells provide protection from infection at barrier sites. In the small intestine, T_RM cells are found in at least two distinct subpopulations: one with higher expression of effector molecules and another with greater memory potential¹. However, the origins of this diversity remain unknown. Here we proposed that distinct tissue niches drive the phenotypic heterogeneity of T_RM cells. To test this, we leveraged spatial transcriptomics of human samples, a mouse model of acute systemic viral infection and a newly established strategy for pooled optically encoded gene perturbations to profile the locations, interactions and transcriptomes of pathogen-specific T_RM cell differentiation at single-transcript resolution. We developed computational approaches to capture cellular locations along three anatomical axes of the small intestine and to visualize the spatiotemporal distribution of cell types and gene expression. Our study reveals that the regionalized signalling of the intestinal architecture supports two distinct T_RM cell states: differentiated T_RM cells and progenitor-like T_RM cells, located in the upper villus and lower villus, respectively. This diversity is mediated by distinct ligand–receptor activities, cytokine gradients and specialized cellular contacts. Blocking TGFβ or CXCL9 and CXCL10 sensing by antigen-specific CD8 T cells revealed a model consistent with anatomically delineated, early fate specification. Ultimately, our framework for the study of tissue immune networks reveals that T cell location and functional state are fundamentally intertwined.