Molecular features of response and resistance to glofitamab, a T-cell engager for treatment of large B-cell lymphoma.

T-cell engagers (TCEs) have recently transformed the therapeutic landscape of hematological malignancies, including relapsed or refractory (R/R) B-cell non-Hodgkin lymphoma (B-NHL). However, the variability in patient responses underscores the need for a deeper mechanistic understanding of the factors driving efficacy. Immune cell composition and T-cell functional states are emerging as critical determinants of immunotherapy outcomes. Recent advances in single-cell RNA sequencing (scRNA-seq) technologies have enabled high-resolution characterization of T-cell states, revealing a spectrum from highly activated effectors to exhausted or dysfunctional subsets within the tumor microenvironment. In this study, we conducted longitudinal scRNA-seq analyses and functional assessments of peripheral blood immune cells (peripheral blood mononuclear cells [PBMCs]) from patients with glofitamab-treated R/R B-NHL, achieving complete metabolic response, or with progressive metabolic disease. Our findings reveal that the maintenance of naïve-like ("fresher") T-cell states (particularly the fresher cytotoxic T cells) at early time points is associated with clinical efficacy. In line with molecular data, T cells from responders exhibited enhanced functional activity compared with nonresponders. Furthermore, the analysis of patient PBMCs and intratumor T cells from preclinical tumor models after consecutive glofitamab treatments revealed sustained functional activity, underscoring the long-term durability of T-cell responses. Combination of glofitamab with 4-1BB costimulation translated into increased proportions of intratumor T cells having a fresher, naïve-like phenotype, ultimately leading to stronger antitumor efficacy. Taken together, our findings underscore the therapeutic relevance of fresher, naïve-like T-cell states and the potential of leveraging 4-1BB costimulation to overcome TCE resistance and improve clinical responses in aggressive lymphomas.