Shukla Divanshu, Manne Sasikanth, Jiang Shuguang, Ruella Marco, Wherry E John, Riley James L
Department of Microbiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Institute for Immunology and Immune Health, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
Institute for Immunology and Immune Health, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
Mol Ther. 2025 Aug 5. doi: 10.1016/j.ymthe.2025.08.002.
We explore whether T cells expressing two chimeric antigen receptors (CARs) with distinct signaling motifs (dual CARs) improve CART cell (CART) efficacy against leukemia and lymphoma. Moreover, we investigate whether dual CARTs targeting two antigens (multi-targeted) are superior to dual CARTs targeting a single antigen (single targeted). Functional assays revealed that multi-targeted dual CARTs targeting both CD19 and CD22 were more potent than single-targeted dual CARTs targeting only CD19 or CD22. RNA expression profiling demonstrated that single-targeted dual CARTs augmented canonical nuclear factor κB (NF-κB), non-canonical NF-κB, and Th17 differentiation pathways equivalently upon target engagement. Interestingly, the transcriptional profile of multi-targeted dual CARTs favored the co-stimulatory domain linked to the binder of the more robustly tumor-expressed CD19 rather than one linked to the binder of the less tumor-expressed CD22. In vivo and T cell exhaustion assays found that multi-targeted dual CARTs led to greater durable control of B-ALL than single-targeted dual CARTs, with T cells co-expressing CD19.BBζ and CD22.28ζ being the most potent. These data indicate that optimal pairing of CAR binder domain with signaling cassette bolsters anti-tumor efficacy.
我们探究表达具有不同信号基序的两种嵌合抗原受体（CARs）的T细胞（双特异性CAR T细胞）是否能提高CAR T细胞（CART）对抗白血病和淋巴瘤的疗效。此外，我们研究靶向两种抗原的双特异性CART细胞（多靶点）是否优于靶向单一抗原的双特异性CART细胞（单靶点）。功能分析显示，靶向CD19和CD22的多靶点双特异性CART细胞比仅靶向CD19或CD22的单靶点双特异性CART细胞更有效。RNA表达谱分析表明，单靶点双特异性CART细胞在与靶点结合后同等程度地增强了经典核因子κB（NF-κB）、非经典NF-κB和Th17分化途径。有趣的是，多靶点双特异性CART细胞的转录谱有利于与肿瘤表达更强烈的CD19结合剂相连的共刺激结构域，而不是与肿瘤表达较弱的CD22结合剂相连的共刺激结构域。体内和T细胞耗竭试验发现，多靶点双特异性CART细胞比单靶点双特异性CART细胞能更持久地控制B-ALL，其中共表达CD19.BBζ和CD22.28ζ的T细胞最为有效。这些数据表明，CAR结合结构域与信号盒的最佳配对可增强抗肿瘤疗效。
