纳米抗体靶向 MSLN CAR-T 细胞治疗在转移性实体瘤中抗肿瘤效应的时序图谱

Studies on the dynamic changes occurring in the tumor microenvironment (TME) following CAR-T cell therapy have been confounded by host lymphodepletion, multiple dosing and immunodeficient models.

Here, a nanobody-based, mouse mesothelin-targeting CAR-T cell (A101) was developed, achieving effective primary tumor suppression, metastasis reduction, and improved survival after a single dose in immunocompetent, syngeneic mouse models without lymphodepletion. Temporal tumor profiling using RNA sequencing revealed initial downregulation of cell proliferation genes followed by upregulation of inflammation, epithelial-to-mesenchymal-transition (EMT) and extracellular matrix (ECM) modification genes in the CAR-T-treated tumors relative to mock-T-treated controls.

This phenotype was reversed at a later timepoint which coincided with downregulation of immunosuppressive Cd274 + Lcn2 + neutrophils and upregulation of anti-tumor P2rx1 + Nrf2 - neutrophils. At the same time, upregulation of Ccl2 + in fibroblasts and a more immunomodulatory macrophage phenotype was observed in CAR-T-treated tumors, indicating a tumor adaptation mechanism.

This study demonstrates complex dynamic changes in the TME, and highlights time-dependent responses of solid tumors to CAR-T cell therapy. It further highlights Lcn2 + neutrophils and Ccl2 + fibroblasts as potential therapeutic targets for improving CAR-T cell anti-tumor efficacy.