GEOapplication/xmlftp://ftp.ncbi.nlm.nih.gov/geo/series/GSE313nnn/GSE313648/primaryOK200TranscriptomicsHomo sapiens OtherExpression profiling by high throughput sequencinghttps://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE313648GEOGSEfalseMultimodal profiling of Her2/neu-specific TCR-T cells reveals clonotype-dependent activation programs underlying differential cytotoxic efficacyAdoptive T cell therapy targeting tumor-associated antigens offers a promising avenue for cancer immunotherapy, yet the mechanisms underlying functional heterogeneity among T cell receptors (TCR) recognizing the same antigen remain poorly understood. Here, we comprehensively profile three Her2/neu-specific TCR constructs using integrated multimodal analysis that combines cytotoxicity assays, single-cell multiomics, bulk transcriptomics, cytokine profiling, and in vivo tumor modeling. Despite identical antigen specificity, the constructs exhibit a functional hierarchy: Construct #3 displays potent cytotoxicity in vitro and potent tumor regression in vivo, whereas Constructs #1 and #2 were less effective. Single-cell proteotranscriptomic analysis shows that tumor cell encounter drives activation of cytotoxic CD8⁺ T effector cells and transdifferentiation of NKT cells into APC-like cells. Interactomics also revealed high communication probability between APC-like NKT cells and CD4⁺ T cells, potentially forming a cooperative activation network that sustains effector function. Bulk transcriptomic and secretomic profiling showed that Construct #3 couples cytolytic gene expression with the secretion of anti-tumor cytokines and effector molecules. Together, these findings establish a mechanistic link between anti-Her2/neu TCR-driven transcriptional programming and therapeutic efficacy, illustrating how multi-omic approaches can inform the rational design of potent TCR therapies for solid tumors.2026/05/04GSE313648GSM937170234281313648Homo sapiens