LAYN在肝细胞肝癌中介导肿瘤特异性CTL耗竭的研究

前期我们对肝癌病人5063个T淋巴细胞进行了深度单细胞转录组测序，已鉴定出与T细胞功能耗竭相关的潜在新基因LAYN。该基因在耗竭型CD8+T淋巴细胞中表达显著升高。据此提出假说：LAYN是导致CD8+T细胞功能耗竭的新基因，LAYN表达增高可导致CD8+T淋巴细胞功能耗竭，从而抑制其抗肿瘤免疫反应。本课题中，拟研究LAYN基因能否导致肿瘤抗原特异性CD8+T淋巴细胞的表型、增殖和杀伤功能、各负性调控和转录因子的表达及DNA甲基化水平等特征，向功能耗竭状态转化；通过敲除LAYN基因或通过RNA干扰抑制LAYN基因的表达是否能够逆转T细胞功能耗竭状态，并增强其对肿瘤的杀伤功能。这将有助于深入了解T淋巴细胞耗竭的机制，并有助于将LAYN作为潜在干预靶点，为原发性肝癌患者提供一种新的免疫治疗方法。

In previous studies we performed deep single-cell RNA sequencing on 5063 single T lymphocytes isolated from patients with hepatocellular carcinoma and identified a potential novel T cell exhaustion-related gene LAYN, which was highly expressed in exhausted CD8+T lymphocytes. We hypothesize that LAYN is a new exhaustion-related gene, and its increasing expression may lead to the functional exhaustion of CD8+T cells, thereby inhibiting their anti-tumor immune response. In this study, we will explore whether the increased expression of LAYN could induce the characteristics of tumor antigen-specific T lymphocytes, including their phenotypes, proliferation and killing abilities, expression of various negative regulatory and transcription factors, and DNA methylation level, into functional exhaustion status. By knocking out LAYN gene, or inhibiting LAYN expression with SiRNA, we will further investigate whether these procedures can reverse T cell’s exhaustion status and promote their anti-tumor abilities. This study will provide deep insight into the mechanism of T lymphocyte exhaustion and help us to develop new immunotherapeutic strategy for patients with primary liver cancer by using the LAYN pathway as potential target.