Dasatinib-resistant universal CAR-T cells proliferate in the presence of host immune cells and exhibit antitumor activity

作者：Yuhang Cheng, Jiayuan Zhang, Wei Mu, Shanwei Ye, Jiali Cheng, Zhu Li, Gaoxiang Wang, Yang Cao, Dengju Li, Guang Hu, Liang Huang, Jue Wang, Jianfeng Zhou · 发表于：Molecular Therapy · 年份：2025 · DOI：10.1016/j.ymthe.2025.02.012 · 被引用次数：9 · 研究领域：CAR-T cell therapy research、Immune Cell Function and Interaction

The universal chimeric antigen receptor T cell (UCAR-T) immunotherapy derived from healthy donors holds great promise in pan-cancer treatment. However, UCAR-T cell therapy faces a challenge in the rapid elimination of allogeneic cells by the host immune system. To address this, we introduced a T316I mutation in the leukocyte-specific protein tyrosine kinase (LCK) locus in CAR-T cells using the cytosine base editor (CBE) system. Concurrently, we disrupted endogenous T cell receptor alpha chain (TRAC) and beta-2 microglobulin (B2M) with the CRISPR-Cas9 system, along with dasatinib to overcome host immune rejection, an Src family kinase (SFK) inhibitor. The resulting LCK mutated UCAR-T (KM UCAR-T) cells exhibited normal phenotypes in activation, proliferation, differentiation, and tumor cytotoxicity in vitro. Moreover, KM UCAR-T cells demonstrated sustained expansion in mixed lymphocyte reactions (MLR) when incubated with T cells or peripheral blood mononuclear cells (PBMCs) from HLA-mismatched donors upon dasatinib treatment. Additionally, we illustrated that KM UCAR-T cells displayed antitumor activity in a xenograft murine model and verified the expansion and cytotoxicity of KM UCAR-T over traditional UCAR-T in the presence of allogeneic PBMCs when treated with dasatinib in vivo. These findings offer a novel strategy for UCAR-T cells to resist host immune rejection and achieve sustained expansion.