Yi Yan, Chai Xiaoshan, Zheng Liping, Zhang Yongjing, Shen Jiankai, Hu Biliang, Tao Guangshi
The Second Xiangya Hospital of Central South University, Changsha, Hunan, 410011, China.
Hunan Key Laboratory of Tumor Models and Individualized Medicine, Changsha, Hunan, 410011, China.
Cell Discov. 2021 Apr 27;7(1):27. doi: 10.1038/s41421-021-00255-4.
Revolutionary CART therapy still faces the challenge of severe cytokine release syndrome (CRS). While IL6 and IL1 have been demonstrated as essential contributors, GM-CSF is one of the most abundant inflammatory cytokines secreted by CART and has also been suggested in contributing to CRS. To minimize GM-CSF production from CART to reduce its associated toxicity, we conducted a pilot study (ChiCTR2000032124) of CRISPR-edited GM-CSF knockout (KO) in CART secreting anti-IL6 scFv and IL1RA, with additional TCR KO for tracing edited CART. The initial results of three patients (1 Non-Hodgkin lymphoma (NHL) and 2 multiple myelomas (MMs)) are summarized as: 3/3 complete response, 2/3 none CRS, 1/3 grade 2 CRS, and 0/3 neurotoxicity. The analysis revealed low levels of GM-CSF, IL6 and IL1B at the time of interferon-gamma (IFNG) peaks, and elevated IL1RA. We also observed significant expansion of CD3 CART during treatment and no aberrant expansion of CD3 CART in the follow-up. Re-expansion of CD3 CART was observed in two patients while recurring CD19 cells were eradicated in the patient with NHL. In summary, our study supported the safety and durable potency of CRISPR-edited CART in patients, providing a novel platform for developing autologous or allogeneic CART to minimize GM-CSF-associated toxicity in addition to autonomous IL6/IL1 blockade.
革命性的嵌合抗原受体T细胞（CART）疗法仍面临严重细胞因子释放综合征（CRS）的挑战。虽然白细胞介素6（IL6）和白细胞介素1（IL1）已被证明是CRS的重要促成因素，但粒细胞-巨噬细胞集落刺激因子（GM-CSF）是CART分泌的最丰富的炎性细胞因子之一，也被认为与CRS有关。为了尽量减少CART产生的GM-CSF以降低其相关毒性，我们开展了一项初步研究（ChiCTR2000032124），对分泌抗IL6单链抗体片段（scFv）和IL1受体拮抗剂（IL1RA）的CART进行CRISPR编辑的GM-CSF基因敲除（KO），并额外进行T细胞受体（TCR）基因敲除以追踪编辑后的CART。三名患者（1例非霍奇金淋巴瘤（NHL）和2例多发性骨髓瘤（MM））的初步结果总结如下：3/3完全缓解，2/3无CRS，1/3为2级CRS，0/3无神经毒性。分析显示，在干扰素-γ（IFNG）峰值时GM-CSF、IL6和白细胞介素1β（IL1B）水平较低，而IL1RA升高。我们还观察到治疗期间CD3 CART显著扩增，随访中CD3 CART无异常扩增。两名患者观察到CD3 CART再次扩增，而NHL患者中复发的CD19细胞被清除。总之，我们的研究支持了CRISPR编辑的CART在患者中的安全性和持久效力，除了自主阻断IL6/IL1外，还为开发自体或异体CART以尽量减少GM-CSF相关毒性提供了一个新平台。
