免疫治疗在肝癌中的应用已初现端倪，但如何发现抗原靶点并调动免疫反应仍然值得探究。在本研究中，申请人基于前期利用免疫多肽组学展示MHC-I抗原呈递谱以及肝癌蛋白基因组学数据集的基础上，全面表征了肝癌的肿瘤-睾丸（CT）抗原谱特征，并发现CT抗原POTEE基因在肝癌中特异性高表达且多次被MHC-I所呈递，在后续研究中将进一步探索通过MHC-I对其呈递的多肽谱、寻找潜在免疫原性多肽、抗原体外刺激T细胞并筛选特异性TCR基因座、CRISPR工程化改造生成POTEE特异性T细胞并进行功能验证。并将探究POTEE抗原暴露是否可间接增强肿瘤免疫原性成为免疫“热肿瘤”，以便于与PD-1等现有免疫治疗的联合策略。本课题期望构建可惠及30%以上POTEE阳性肝癌患者的TCR-T细胞免疫治疗管线，并解析POTEE功能调节机制及免疫原性形成机制，为肝癌个体化细胞免疫治疗提供基础。

The application of immunotherapy in liver cancer has begun to emerge, but how to find antigen targets and mobilize the immune response is still worth exploring. In this study, we has fully characterized the cancer-testis (CT) antigen profile of liver cancer, based on the major histocompatibility complex-I (MHC-I) immunopeptideomics and proteogenomics database to display the specific CT antigen presentation spectrum in liver cancer, and found that the gene POTEE is specifically overexpressed in liver cancer and presented by MHC-I. In subsequent studies, we will further explore the peptide profile presented by MHC-I, search potential immunogenic peptides, stimulate T cells to expand by peptides in vitro, and generate POTEE-specific T cells by specific TCR loci. We will also explore whether the exposure of POTEE antigen can indirectly enhance tumor immunogenicity and become an immune "hot tumor", so as to facilitate the combined strategy with existing immunotherapy such as PD-1. We expect to construct a TCR-T cell immunotherapy pipeline that can benefit more than 30% of patients with POTEE-positive liver cancer, and analyze the mechanism of POTEE regulation and immunogenicity mechanism to provide the basis for individualized cell immunotherapy.