Zhang Chenglin, Liu Yanyan, Guo Haoran, Peng Ying, Huang Lei, Lu Shuangshuang, Wang Zhimin
National Center for International Research in Cell and Gene Therapy, Sino-British Research Centre for Molecular Oncology, School of Basic Medical Sciences, Academy of Medical Sciences, Zhengzhou University, Zhengzhou 450052, China.
Department of Internal Medicine, Affiliated Cancer Hospital of Zhengzhou University & Henan Cancer Hospital, Zhengzhou 450052, China.
Biomedicines. 2025 Jun 30;13(7):1598. doi: 10.3390/biomedicines13071598.
Esophageal squamous cell carcinoma (ESCC) is a common form of esophageal cancer with a poor prognosis and limited treatment options. Epidermal growth factor receptor (EGFR), an overexpressed oncogenic gene in all ESCC patients, is an attractive target for developing therapies against ESCC. There is an extremely urgent need to develop immunotherapy tools targeting EGFR for the treatment of ESCC. In this study, we developed human Interleukin-21 (hIL-21)-armed, chimeric-antigen-receptor-modified T (CAR-T) cells targeting EGFR as a new therapeutic approach. The CAR contains a variable domain of the llama heavy chain of heavy-chain antibodies (VHHs), also known as nanobodies (Nbs), as a promising substitute for the commonly used single-chain variable fragment (ScFv) for CAR-T development. We show that nanobody-derived, EGFR-targeting CAR-T cells specifically kill EGFR-positive esophageal cancer cells in vitro and in animal models. Human IL-21 expression in CAR-T cells further improved their expansion and antitumor ability and were observed to secrete more interferon-gamma (IFN-γ), tumor necrosis factor alpha (TNF-α), and Interleukin-2 (IL-2) when co-cultured with ESCC cell lines in vitro. More CD8 CAR-T cells and CD3CD8CD45ROCD62L central memory T cells were detected in CAR-T cells expressing hIL-21 cells. Notably, hIL-21-expressing CAR-T cells showed superior antitumor activity in vivo in a KYSE-150 xenograft mouse model. Our results show that hIL-21-armed, nanobody-derived, EGFR-specific CAR-T cell therapy is a highly promising option for treating ESCC patients.
食管鳞状细胞癌（ESCC）是食管癌的一种常见形式，预后较差且治疗选择有限。表皮生长因子受体（EGFR）在所有ESCC患者中均为过度表达的致癌基因，是开发针对ESCC治疗方法的一个有吸引力的靶点。迫切需要开发针对EGFR的免疫治疗工具来治疗ESCC。在本研究中，我们开发了靶向EGFR的人白细胞介素-21（hIL-21）武装的嵌合抗原受体修饰T（CAR-T）细胞作为一种新的治疗方法。该CAR包含重链抗体（VHHs）的羊驼重链可变结构域，也称为纳米抗体（Nbs），作为CAR-T开发中常用的单链可变片段（ScFv）的一种有前景的替代物。我们表明，源自纳米抗体的靶向EGFR的CAR-T细胞在体外和动物模型中能特异性杀伤EGFR阳性食管癌细胞。CAR-T细胞中hIL-21的表达进一步提高了它们的扩增和抗肿瘤能力，并且在体外与ESCC细胞系共培养时观察到它们分泌更多的干扰素-γ（IFN-γ）、肿瘤坏死因子-α（TNF-α）和白细胞介素-2（IL-2）。在表达hIL-21的CAR-T细胞中检测到更多的CD8 CAR-T细胞和CD3CD8CD45ROCD62L中央记忆T细胞。值得注意的是，表达hIL-21的CAR-T细胞在KYSE-150异种移植小鼠模型中在体内表现出卓越的抗肿瘤活性。我们的结果表明，hIL-21武装的、源自纳米抗体的、EGFR特异性CAR-T细胞疗法是治疗ESCC患者的一个极有前景的选择。
