Yi Fei, Cohen Tal, Zimmerman Natalie, Dündar Friederike, Zumbo Paul, Eltilib Razan, Brophy Erica J, Arkin Hannah, Feucht Judith, Gormally Michael V, Hackett Christopher S, Kropp Korbinian N, Etxeberria Inaki, Chandran Smita S, Zhao Zeguo, Cai Winson, Daniyan Anthony F, Park Jae H, Lareau Caleb A, Hsu Katharine C, Sadelain Michel, Betel Doron, Klebanoff Christopher A
Immuno-Oncology Program, Memorial Sloan Kettering Cancer Center (MSKCC), New York, NY, USA.
Department of Pediatric Hematology/Oncology, Weill Cornell Medical College, New York, NY, USA.
Nat Cancer. 2025 Jul 22. doi: 10.1038/s43018-025-01009-x.
Chimeric antigen receptor (CAR)-engineered lymphocytes treat B cell malignancies; however, limited persistence can restrain the full therapeutic potential of this approach. FAS ligand (FAS-L)/FAS interactions govern lymphocyte homeostasis. Knowledge of which cells express FAS-L in patients with cancer and whether these sources compromise CAR persistence remains incomplete. Here, we constructed a single-cell atlas of diverse cancers to identify cellular subsets expressing FASLG, the gene encoding FAS-L. We discovered that FASLG expression is limited primarily to endogenous T cells, natural killer (NK) cells and CAR-T cells, while tumor and stromal cell expression is minimal. To establish whether CAR-T and CAR-NK cell survival is FAS-L regulated, we performed competitive fitness assays using FAS-dominant negative receptor (ΔFAS)-modified lymphocytes. Following transfer, ΔFAS-expressing CAR-T/CAR-NK cells became enriched, a phenomenon that mechanistically was reverted through FASLG knockout. By contrast, FASLG was dispensable for CAR-mediated tumor killing. In multiple models in female mice, ΔFAS coexpression enhanced antitumor efficacy. Together, these findings reveal that CAR-engineered lymphocyte persistence is governed by a FAS-L/FAS autoregulatory circuit.
嵌合抗原受体（CAR）工程化淋巴细胞可治疗B细胞恶性肿瘤；然而，有限的持久性会限制这种方法的全部治疗潜力。FAS配体（FAS-L）/FAS相互作用控制淋巴细胞的稳态。关于癌症患者中哪些细胞表达FAS-L以及这些来源是否会影响CAR的持久性，目前仍不完全清楚。在这里，我们构建了一个多种癌症的单细胞图谱，以识别表达FASLG（编码FAS-L的基因）的细胞亚群。我们发现FASLG的表达主要限于内源性T细胞、自然杀伤（NK）细胞和CAR-T细胞，而肿瘤细胞和基质细胞的表达极少。为了确定CAR-T和CAR-NK细胞的存活是否受FAS-L调节，我们使用FAS显性负性受体（ΔFAS）修饰的淋巴细胞进行了竞争性适应性分析。转移后，表达ΔFAS的CAR-T/CAR-NK细胞富集，这一现象通过FASLG基因敲除在机制上得以逆转。相比之下，FASLG对于CAR介导的肿瘤杀伤是可有可无的。在雌性小鼠的多个模型中，共表达ΔFAS增强了抗肿瘤疗效。总之，这些发现揭示了CAR工程化淋巴细胞的持久性受FAS-L/FAS自调节回路的控制。
