NF2肿瘤新抗原特异性TCR挖掘及其介导的肿瘤杀伤活性研究

2型神经纤维瘤病（NF2）为良性多发肿瘤综合征，具有恶性生物学特征，手术不能根治且预后差。新抗原特异TCR基因修饰T细胞（TCR-T）可实现对肿瘤细胞的靶向识别及特异杀伤，并有望扩展运用于各种实体瘤的临床治疗，但在NF2中未见报道。我们前期在1例NF2患者听神经瘤中鉴定出2个肿瘤新抗原，进一步通过测序，鉴定并构建的新抗原特异性TCR-T体外可分泌高水平IFN-γ，提示其应用于NF2治疗的可能。故本项目拟在10例NF2患者中：1利用全外显子及转录组测序技术筛选，细胞共培养实验验证，进一步挖掘NF2肿瘤新抗原；2通过流式技术分选新抗原特异反应性TCR，免疫组库及单细胞测序获得匹配的α及β链序列，构建TCR-T细胞；3研究TCR-T体外杀伤NF2肿瘤细胞的能力及可能机制，同时在移植瘤小鼠模型中评价体内杀伤肿瘤效果。该项目的完成有助于NF2有效免疫治疗方案的确立，并为神经系统肿瘤治疗提供新的思路。

Neurofibromatosis type 2 (NF2) is a systemic multiple benign tumor syndrome with malignant biological characteristics, surgery can not cure it and the prognosis is poor. Neoantigen-specific TCR-engineered T cells (TCR-T), which is expected to cure all kinds of solid tumors, could specifically target and kill the tumor cells. However, there is no report in NF2. In the previous study, we identified 2 tumor neoantigen in NF2-associated schwannoma, and further we construct these neoantigen-specific TCR-T through sequencing and identified one could secret high.level of IFN-γ, suggesting the possibility of its application to NF2 treatment. So in the current project, we will study the following points in 10 NF2 patients: 1 ,Identify the NF2 tumor neoantigen through mining exomic and transcriptome sequencing data and then validate through cell co-culture experiments; 2 ,Sort the neoantigen-specific TCR using FACS, and obtain α and β chain sequence using immune repertoire and single cell sequencing and finally construct the TCR-T cells; 3, Study the cytotoxic potential and probable mechanisms of TCR-T against.NF2-associated tumor cells in vitro, and then investigate the anti-tumor effect in mice xenotransplant model. The completion of the project is conducive to the establishment of an effective NF2 immunotherapy scheme, and provides a new way of thinking for the treatment of nervous system tumors.