scRNA+ TCR-seq revealed dual TCR T cells antitumor response in the TME of NSCLC

Abstract

The intricate origins, subsets, and characteristics of TCR (T Cell Receptor) s, along with the mechanisms underpinning the antitumor response of tumor-infiltrating T lymphocytes within the tumor microenvironment (TME) remain enigmatic. Recently, the advent of single-cell RNA+TCR-sequencing (scRNA+TCR seq) has revolutionized TME analysis, providing unprecedented insight into the origins, cell subsets, TCR CDR3 compositions, and the expression patterns of response/depletion factors within individual tumor-infiltrating T lymphocytes. Our analysis of the shared scRNA+TCR seq dataset revealed a substantial presence of dual TCR T cells, characterized by clonal hyperplasia and remarkable migratory prowess across various tissues, including blood, normal, peritumoral, and tumor tissues in non-small cell lung cancer patients. Notably, dual TCR CD8+T cells predominantly fell within the CXCL13+subset, displaying potent antitumor activity and a strong preference for tumor tissue residency. Conversely, dual TCR CD4+T cells were predominantly classified as CD5+ or LMNA+subsets, exhibiting a more even distribution across diverse tissue types. By harnessing scRNA+TCR seq and other cutting-edge technologies, we can delve deeper into the effects and mechanisms that regulate the antitumor response or tolerance of dual TCR T cells. This innovative approach holds immense promise in offering fresh perspectives and avenues for advancing research on TIL (Tumor infiltrating lymphocyte)s within the TME.

Keywords: Lung Cancer; T cell Receptor - TCR; Tumor infiltrating lymphocyte - TIL; Tumor microenvironment - TME.

Figure 1. Single-cell RNA+TCR-sequencing (scRNA+TCR seq) revealed dual TCR T cells antitumor response in the TME of NSCLC. (A) Dual TCR CD4+T and dual TCR CD8+T ratios in blood, normal, peri, and tumor tissue of each patient. (B) Dual TCR CD4+T CDR3 overlap and dual TCR CD8+T CDR3 overlap in blood, normal, peri, tumor tissue of each patient. (C) TRAV, TRAJ, TRBV, TRBJ usage of single TCR T cells and dual TCR T cells. (D) Dual TCR T cells accounted for the proportion of total dual TCR T cells in blood, normal, peri, and tumor tissue. (E) Ratios of dual TCR CD4+T and dual TCR CD8+T in each subset. (F) Single TCR T and dual TCR T diversity (1/DS). (G) Single TCR T cells and dual TCR T cells clonality (clone count:2–5, 6–9,≥10). (Original clones proliferating to cell numbe r≥2 in online supplemental table 2), including ‘single TCR T cell+dual TCR T cell’ ≥2 clones. For instance, P1_TCR_286 clone type (5 cells) in CD4+T cells in online supplemental table 2)=single TCR CD4+T (1 cell)+dual TCR CD4+T (4 cells), but in our analysis, single TCR and dual TCR were separated for comparative analysis. Therefore, single TCR or dual TCR T with cell number 1 was not analyzed in clonal amplification analysis.) (H) Number, sequence composition, source patient and tissue, and subsets of dual TCR CD4+T and dual TCR CD8+T from the top five high-frequency clones. TCR, T Cell Receptor; TRAV, T cell receptor alpha chain Variable gene; TRAJ, T cell receptor alpha chain Jointing gene; TRBV, T cell receptor beta chain Variable gene; TRBJ, T cell receptor beta chain Jointing gene; NSCLC, non-small cell lung cancer; TME, tumor microenvironment.