Table_1_Tenascin-C can Serve as an Indicator for the Immunosuppressive Microenvironment of Diffuse Low-Grade Gliomas.doc

PurposeThe development and progression of glioma are associated with the tumor immune microenvironment. Diffuse low-grade gliomas (LGGs) with higher immunosuppressive microenvironment tend to have a poorer prognosis. The study aimed to find a biological marker that can reflect the tumor immune microenvironment status and predict prognosis of LGGs.
MethodsThe target gene tenascin-C (TNC) was obtained by screening the Chinese Glioma Genome Atlas (CGGA) and the Cancer Genome Atlas (TCGA) databases. Then samples of LGGs were collected for experimental verification with immunohistochemistry, immunofluorescence, immunoblotting, quantitative real-time PCR. ELISA was employed to determine the content of TNC in serum and examine its relationship with the tumor immune microenvironment. Eventually, the sensitivity of immunotherapy was predicted on the basis of the content of TNC in LGGs.
ResultsIn the high-TNC subgroup, the infiltration of immunosuppressive cells was increased (MDSC: r=0.4721, Treg: r=0.3154, etc.), and immune effector cells were decreased [NKT, γδT, etc. (p<0.05)], immunosuppressive factors were elevated [TGF-β, IL10, etc. (p<0.05)], immunostimulatory factors, such as NKG2D, dropped (p<0.05), hypoxia scores increased (p<0.001), and less benefit from immunotherapy (p<0.05). Serum TNC level could be used to assess the status of tumor immune microenvironment in patients with grade II (AUC=0.8571; 95% CI: 0.6541-1.06) and grade III (AUC=0.8333; 95% CI: 0.6334-1.033) glioma.
ConclusionsOur data suggested that TNC could serve as an indicator for the immunosuppressive microenvironment status and the prognosis of LGGs. Moreover, it could also act as a predictor for the effect of immunotherapy on LGG patients.

### 研究目的
胶质瘤的发生发展与肿瘤免疫微环境（tumor immune microenvironment）密切相关。免疫抑制微环境程度更高的弥漫性低级别胶质瘤（Diffuse low-grade gliomas, LGGs）往往预后较差。本研究旨在筛选可反映肿瘤免疫微环境状态、并能预测LGGs患者预后的生物学标志物。

### 研究方法
本研究通过筛选中国胶质瘤基因组图谱（Chinese Glioma Genome Atlas, CGGA）与癌症基因组图谱（Cancer Genome Atlas, TCGA）数据库，筛选得到目标基因腱生蛋白C（tenascin-C, TNC）。随后收集LGGs患者样本，采用免疫组化、免疫荧光、免疫印迹、定量实时PCR进行实验验证；通过酶联免疫吸附试验（ELISA）检测血清中TNC的含量，并分析其与肿瘤免疫微环境的相关性。最终基于LGGs组织中TNC的表达水平预测免疫治疗敏感性。

### 研究结果
在TNC高表达亚组中，免疫抑制细胞浸润水平升高（髓系来源抑制细胞（MDSC）：r=0.4721，调节性T细胞（Treg）：r=0.3154 等），免疫效应细胞浸润水平降低[自然杀伤T细胞（NKT）、γδT细胞等，p<0.05]；免疫抑制因子表达上调[转化生长因子-β（TGF-β）、白细胞介素10（IL10）等，p<0.05]，免疫刺激因子如自然杀伤细胞2族成员D（NKG2D）表达下调（p<0.05）；缺氧评分升高（p<0.001），患者从免疫治疗中获益更少（p<0.05）。血清TNC水平可用于评估Ⅱ级（AUC=0.8571；95%置信区间CI：0.6541~1.06）与Ⅲ级（AUC=0.8333；95%CI：0.6334~1.033）胶质瘤患者的肿瘤免疫微环境状态。

### 研究结论
本研究数据表明，TNC可作为评估LGGs免疫抑制微环境状态与患者预后的指标，同时也可作为预测LGG患者免疫治疗疗效的预测因子。