一种靶向CAT/ERp57的CAR-NK细胞的抗肿瘤作用研究

嵌合抗原受体修饰的自然杀伤细胞（CAR-NK）是一种新型的肿瘤过继免疫疗法，由于其具备CAR-T没有的诸多优点而备受关注。然而，实体瘤组织浸润差和CAR-NK靶向的肿瘤抗原非特异性表达导致CAR-NK在实体瘤的治疗效果不佳及副作用大，阻碍了其在临床上的应用。寻求更好的CAR-NK肿瘤靶向抗原和提升其对实体瘤的浸润能力是CAR-NK研究的重要问题。研究表明，一些化疗药低剂量不仅可以诱导抗肿瘤微环境同时也能诱导癌细胞发生免疫原性细胞死亡(ICD)，导致原本在内质网表达的多种蛋白转位至癌细胞膜表面，本课题旨在探索ICD转位至癌细胞表面的CRT和ERp57可否作为CAR-NK的靶标？拟进行如下研究：1)筛选鉴定靶向CRT和ERp57的纳米抗体，2）构建基于纳米抗体的CAR-NK-92细胞，3）通过体内外实验评估诱导ICD后其抗肿瘤效果。本研究将为新型CAR-NK过继疗法研发提供理论和实验依据。

Chimeric antigen receptor engineered NK cells (CAR-NK) is a new type of tumor adoptive immunotherapy, which has attracted much attention owing to its advantages that CAR-T do not have. However, the low tissue infiltration of CAR-NK in solid tumors and the non-specific expression of tumor-associated antigens targeted by CAR-NK lead to poor therapeutic effects of CAR-NK in solid tumors and large side effects, which hinders its clinical application. Searching for better CAR-NK targets and improving its ability to infiltrate into solid tumors are important issues in CAR-NK research. Studies have shown that low dose of some chemotherapeutics can not only induce the anti-tumor microenvironment, but also can result in immunogenic cell death (ICD) in cancer cells, causing a panel of proteins originally expressed in the endoplasmic reticulum to translocate to the cancer cell membrane. Therefore, this study aims to explore whether CRT and ERp57, which translocate to the cancer cell membrane during ICD, can be used as CAR-NK targets. The following studies will be conducted: 1) Screening and identifying nanobodies targeting CRT and ERp57, 2) Constructing CAR-NK-92 cells modified with identified nanobodies, 3) In vivo and in vitro experiments to evaluate the anti-tumor effect after induction of ICD. This project will provide theoretical and experimental basis for the development of a new CAR-NK adoptive therapy.