Figure 4.

Chronically stimulated stem-like CAR-T_RM cells sustain a resident memory phenotype in association with epigenetic reprogramming

(A) Principal-component analysis of ATAC-seq data.

(B) Volcano plot demonstrating differentially accessible peaks between CAR-T_RM and CAR-T_CONV CD8⁺ cells.

(C–G) GSEA showing enrichment of (C) CD69⁺CD103⁺CD8⁺ and (D) CD69⁻CD103⁻CD8⁺ T cell signatures. ATAC-seq tracks of genes associated with (E) resident memory T cells (ITGAE, ITGAV, ENTPD1, IRF4), (F) circulating T cells (KLF2), and (G) stemness (TCF7, IL7R). Differentially accessible regions are highlighted in blue.

(H) M5 CAR T cells were repetitively challenged with AsPC1 cells, and frequencies of resident memory (CD103⁺CD39⁺CD8⁺) and stem-like resident memory (CD103⁺IL7R⁺CD8⁺) T cells are shown over time. Bulk ATAC-seq experiments were performed using manufactured CD8⁺ CAR T cells from n = 2 healthy donors as biological replicates. Restimulation assays were performed with CAR T cells generated from n = 3 distinct healthy subjects, also serving as biological replicates. Results from one representative donor are shown.