Murad John P, Christian Lea, Rosa Reginaldo, Ren Yuwei, Buckley Alyssa J, Lee Eric Hee Jun, Lopez Lupita S, Park Anthony K, Yang Jason, Yamaguchi Yukiko, Trac Candi, Adkins Lauren N, Chang Wen-Chung, Martinez Catalina, June Carl H, Forman Stephen J, Ishihara Jun, Lee John K, Stern Lawrence A, Priceman Saul J
Keck School of Medicine (KSOM)/Norris Center for Cancer Cellular Immunotherapy Research (CCCIR), Division of Medical Oncology, Department of Medicine, Keck School of Medicine of USC, Los Angeles, CA, USA.
Department of Hematology and Hematopoietic Cell Transplantation, City of Hope, Duarte, CA, USA.
Nat Biomed Eng. 2025 Oct 1. doi: 10.1038/s41551-025-01509-2.
Chimeric antigen receptor (CAR)-T cell efficacy in solid tumours is limited due in part to the immunosuppressive tumour microenvironment (TME). To improve antitumour responses, we hypothesized that enabling CAR-T cells to secrete bifunctional fusion proteins consisting of a cytokine modifier such as TGFβ, IL-15 or IL-12, combined with an immune checkpoint inhibitor such as αPD-L1, would provide tumour-localized immunomodulation to improve CAR-T cell functionality. Here we engineer CAR-T cells to secrete TGFβ, IL-15 or IL-12 molecules fused to αPD-L1 scFv and assess in vitro functionality and in vivo safety and efficacy in prostate and ovarian cancer models. CAR-T cells engineered with αPD-L1-IL-12 are superior in safety and efficacy compared with CAR-T cells alone and those engineered with αPD-L1 fused with TGFβ or IL-15. Further, αPD-L1-IL-12 engineered CAR-T cells improve T cell trafficking and tumour infiltration, and localize IFNγ production, TME modulation and antitumour responses, with reduced systemic inflammation-associated toxicities. We believe our αPD-L1-IL-12 engineering strategy presents an opportunity to improve CAR-T cell clinical efficacy and safety across multiple solid tumour types.
嵌合抗原受体（CAR）-T细胞在实体瘤中的疗效有限，部分原因是免疫抑制性肿瘤微环境（TME）。为了提高抗肿瘤反应，我们假设使CAR-T细胞分泌由细胞因子调节剂（如转化生长因子β、白细胞介素-15或白细胞介素-12）与免疫检查点抑制剂（如α程序性死亡受体配体1）组成的双功能融合蛋白，将提供肿瘤局部免疫调节，以改善CAR-T细胞的功能。在这里，我们对CAR-T细胞进行工程改造，使其分泌与α程序性死亡受体配体1单链抗体片段（scFv）融合的转化生长因子β、白细胞介素-15或白细胞介素-12分子，并在前列腺癌和卵巢癌模型中评估其体外功能以及体内安全性和疗效。与单独的CAR-T细胞以及用与转化生长因子β或白细胞介素-15融合的α程序性死亡受体配体1改造的CAR-T细胞相比，用α程序性死亡受体配体1-白细胞介素-12改造的CAR-T细胞在安全性和疗效方面更优越。此外，用α程序性死亡受体配体1-白细胞介素-12改造的CAR-T细胞改善了T细胞的转运和肿瘤浸润，并使干扰素γ的产生、TME调节和抗肿瘤反应局部化，同时降低了全身炎症相关的毒性。我们相信，我们的α程序性死亡受体配体1-白细胞介素-12工程策略为提高CAR-T细胞在多种实体瘤类型中的临床疗效和安全性提供了一个机会。
