髓源抑制细胞在舌鳞癌血管生成中的作用

Myeloid-derived suppressor cells (MDSCs) represent a heterogeneous immature cell population, which expanded in cancer patients or tumor-bearing mice, endowing with suppressive functions on immune surveillance, and promotion on tumor growth and angiogenesis. Our recent data suggest MDSCs dramatic increased in tongue squamous cells carcinoma (TSCC) angiogenesis in 4NQO carcinogen induced mice model. In the present project, firstly,we plan to quantify and correlate the expression of MDSCs with TSCC angiogenesis and clinical prognosis. Secondly,we plan to use 4NQO-induced TSCC mice model, observing the dynamic changes of MDSCs in the progression of TSCC, and analyze its relevance with angiogenesis and survival of tumor-bearing mice by immunofluroscence and FACS; Thirdly, by isolating and culturing MDSCs in vitro, we plan to explore the mechanisms TSCC cells promtes MDSCs migration, survival and inhition on autophagy.Explore the possible role of chemokines such as S100A8 /9,CXCR2, CCR5 and their ligands in vitro chemoattract of MDSCs and furthure confirmed by vivo experiment with according neutralized antibodies and lentiviral shRNAs transfection. Finally, we plan to identify the mechanism of MDSCs promoting TSCC-induced angiogenesis via mediating ADAM8 and ADAM28 with angiogenesis models in vitro and in vivo, and further explore the downstream genes. It is expected to reveal novel molecule mechanism and provide innovative molecular targets which MDSCs promote TSCC-induced angigenesis by finish this project.

髓源性抑制细胞（MDSCs）是一群表型异质的髓系来源细胞，在多种恶性肿瘤患者体内大量扩增，具有免疫抑制、促肿瘤生长和转移等作用。本课题组近期实验表明MDSCs参与小鼠舌鳞状细胞癌血管生成。在挖掘前期基因芯片结果基础上，本项目拟从以下几方面重点进行分子机制研究：(1)检测舌鳞癌标本MDSCs的表达，分析其与血管生成及临床预后的关系；(2)构建4NQO舌鳞癌小鼠模型，免疫荧光、流式分选技术检测MDSCs在舌鳞癌发展过程中的动态改变，并分析其与血管生成、生存的关系；(3)体外分选培养MDSCs，验证舌鳞癌细胞通过CXCR2,CCR5相应配体及S100A8/A9对MDSCs趋化、促增殖及抑制自噬的机制，并通过中和性抗体及慢病毒介导的ShRNA策略进行体内阻断研究；(4)利用体外体内血管生成模型验证MDSCs通过ADAM8及ADAM28促进血管生成机制，并明确其下游血管生成靶基因。

髓源性抑制细胞（MDSCs）是一群表型异质的髓系来源细胞，在多种恶性肿瘤患者体内大量扩增，具有免疫抑制、促肿瘤生长和转移等作用。本课题组通过基因表达芯片发现Tgfbr1/Pten双基因敲除小鼠舌癌相对舌粘膜明显上调的一群基因主要与趋化髓系细胞相关。本项目在挖掘前期基因芯片结果基础上为了明确“MDSCs在口腔鳞状细胞癌发生及血管生成中的作用”这一科学问题，从以下几方面重点进行分子机制研究：(1)建立有随访的口腔鳞癌组织芯片，利用连续切片免疫组织化学染色及数字化病理分析，明确人口腔鳞癌标本MDSCs的表达，分析其与相关调控分子、血管生成及临床预后的关系；(2)利用抗体芯片筛选出小鼠口腔鳞癌髓系趋化因子，通过流式细胞术发现荷瘤小鼠外周免疫器官MDSCs增加。通过体内功能性实验，利用小分子或特异性抗体明确HIF-1α，VEGFA，CCL2,CCL3，CXCL1，免疫检查点分子PD-1,CTLA-4，LAG-3, TIM-3, B7-H3, B7-H4, VISTA及腺苷通路CD73，A2aR对MDSCs趋化至口腔鳞癌肿瘤微环境，MDSCs扩增以及MDSCs抑制分化的作用。(3)明确重要髓系转录因子STAT3及CD47/SIRPα信号轴对MDSCs的调控。(4)明确口腔癌小鼠模型EGFR，STAT3, NOTCH1/HES1抑制后MDSCs及血管生成的改变。截止结题时发表第一或通讯作者SCI论文36篇（包括已接收4篇），全部标注本项目资助，其中第一资助17篇，第二资助14篇，一区SCI论文11篇。