Giudice Anna Maria, Matlaga Stephanie, Roth Sydney L, Pascual-Pasto Guillem, Schürch Patrick M, Rouin Geoffrey, McIntyre Brendan, Grothusen Grant P, Cresswell-Clay Evan, Shraim Rawan, Groff David, Zecchino Vincent, Lombardi Simona, Martinez Daniel, Spruce Lynn A, Brown Elizabeth M, Fazelinia Hossein, Henrickson Sarah E, Nance Jonas, Reynolds C Patrick, Bosse Kristopher R
Division of Oncology and Center for Childhood Cancer Research, Children's Hospital of Philadelphia, Philadelphia, PA 19104, USA.
Department of Biomedical and Health Informatics, Children's Hospital of Philadelphia, Philadelphia, PA 19104, USA.
Sci Transl Med. 2025 Nov 19;17(825):eads4214. doi: 10.1126/scitranslmed.ads4214.
Glypican-2 (GPC2) and the disialoganglioside GD2 are validated CAR T cell targets in neuroblastoma, but durable clinical responses remain limited. This modest chimeric antigen receptor T cell (CAR T cell) efficacy is in part due to suboptimal T cell persistence, antigen down-regulation, and a hostile tumor microenvironment, which includes immune cell-modulating extracellular vesicles (EVs). Neuroblastoma-derived EVs may contain CAR targets or other immunoregulatory elements that can modulate CAR T cell antitumor activity. Thus, we first profiled the surfaceome of neuroblastoma EVs and assessed their impact on both GPC2 and GD2 CAR T cell function. Neuroblastoma EVs displayed GPC2 and GD2, with minimal expression of programmed death-ligand 1 (PD-L1), and were detected in blood from tumor-bearing mice and patients. These EVs directly activated paired CAR T cells, suggesting a role for a peripheral source of CAR antigen. To exploit this therapeutically, we engineered nontumor-derived GPC2+ synthetic EVs (SyntEVs) as CAR T cell enhancers and armored them with either albumin-binding domains or GD2-binding domains. In mice harboring human neuroblastoma cell line-derived or patient-derived xenografts, serial infusion of armored SyntEVs after GPC2 CAR T cells enhanced tumor control by boosting peripheral CAR T cell persistence. Moreover, GD2-targeting SyntEVs decorated low-antigen tumor cells with GPC2, circumventing antigen down-regulation. This SyntEV platform offers a versatile system to address the therapeutic limitations of CAR T cells in solid tumors.
磷脂酰肌醇蛋白聚糖-2（GPC2）和双唾液酸神经节苷脂GD2是经证实的神经母细胞瘤嵌合抗原受体T细胞（CAR T细胞）靶点，但持久的临床反应仍然有限。这种CAR T细胞疗效欠佳部分是由于T细胞持久性欠佳、抗原下调以及恶劣的肿瘤微环境，其中包括可调节免疫细胞的细胞外囊泡（EV）。神经母细胞瘤来源的EV可能含有CAR靶点或其他免疫调节元件，可调节CAR T细胞的抗肿瘤活性。因此，我们首先分析了神经母细胞瘤EV的表面组，并评估了它们对GPC2和GD2 CAR T细胞功能的影响。神经母细胞瘤EV表达GPC2和GD2，程序性死亡配体1（PD-L1）表达极少，在荷瘤小鼠和患者的血液中均可检测到。这些EV直接激活配对的CAR T细胞，提示CAR抗原的外周来源发挥了作用。为了将此用于治疗，我们构建了非肿瘤来源的GPC2+合成EV（SyntEV）作为CAR T细胞增强剂，并用白蛋白结合域或GD2结合域对其进行武装。在携带人神经母细胞瘤细胞系来源或患者来源异种移植物的小鼠中，在GPC2 CAR T细胞后连续输注武装的SyntEV，通过提高外周CAR T细胞持久性增强了肿瘤控制。此外，靶向GD2的SyntEV用GPC2修饰低抗原肿瘤细胞，规避了抗原下调。这个SyntEV平台提供了一个通用系统，以解决CAR T细胞在实体瘤中的治疗局限性。
