肿瘤浸润淋巴细胞（TIL）疗法已成为近来肿瘤免疫治疗的热点，被多个欧美临床中心证实在皮肤黑色素瘤患者中可获得超过40%的客观反应率。在中国相对高发的黏膜黑色素瘤亚型未有相关研究报道和临床突破。在前期研究中，我们的对大规模的口腔黏膜黑色素瘤（OMM）进行了遗传学图谱描绘和分子分型。基于此，本项目拟对不同分子分型的OMM进行免疫谱系分析，通过流式细胞术和单细胞测序技术探讨肿瘤浸润免疫细胞的谱系分群和功能分群，探索可提高TIL体外扩增成功率且缩短扩增时间的优选方案。同时，对比分析不同分子分型OMM病人T细胞的扩增潜能及扩增T细胞与外周血T细胞的克隆库差异，并利用OMM患者组织来源的细胞模型及可表达人源化IL-2的NOG小鼠构建的PDX模型评估OMM特异性T细胞的抗肿瘤有效性，从而探讨TIL疗法的可行性。本项目有望为TIL疗法在OMM及黏膜黑色素瘤的临床转化提供新的理论依据和实验基础。

Multiple clinical trials have proven the effectiveness of Tumor Infiltrating Lymphocytes (TIL) in melanoma patients with more than 40% of overall responsive rate (ORR). However, the breakthrough for TIL therapy in Chinese melanoma patients, especially one major subtype - oral mucosal melanoma (OMM) patients, has yet been reported. Previously, we have characterized the genomic underpinnings of a large group of OMMs and delineated the disease molecular subtypes. However, the detailed immune populations within OMM samples is poorly understood. With the aid of flow-cytometry and single cell sequencing technology, we plan to define the immune cells heterogeneity and the relevant functionality in OMM tumor microenvironment. Moreover, different expansion protocols for TIL culture will be explored to increase manufacturing success rate and shorten the manufacturing time. Through analyzing the proliferation capacity and comparing PBMC and TIL product TCR clonal types, a series of preferred OMM TIL expansion protocols would be selected. The anti-tumor efficacy of expanded T cells will be evaluated in molecularly defined OMM patient derived cells and patient derived xenograft (PDX) models established with humanized IL-2 NOG mice, to explore potential feasibility of TIL therapy in treating OMM. The proposed study would serve as a solid theoretical and practical foundation for the potential TIL treatments in Chinese OMM patients.