SYK影响肿瘤相关巨噬细胞极化促进胆管癌进展的机制研究

胆管细胞癌恶性程度高，肿瘤相关巨噬细胞与其进展密切相关。前期研究发现：大/小鼠胆管癌演变进程中SYK表达增高，伴随着M2型巨噬细胞浸润数目增多；人胆管癌组织中SYK高表达，与肿瘤大小呈正相关，提示患者的预后不良；SYK抑制剂显著延缓裸鼠皮下移植瘤和大鼠胆管癌的肿瘤生长，伴肿瘤M2型巨噬细胞浸润减少。据此推测：SYK通过影响肿瘤相关巨噬细胞向M2型极化促进胆管癌的进展。本项目拟通过体内外实验研究SYK在胆管癌中对肿瘤相关巨噬细胞极化的影响及其生物学功能；利用肝脏原代细胞分离、转录组学测序及生信分析，探索SYK介导肿瘤相关巨噬细胞影响胆管癌发生发展的调控机制；对大/小鼠胆管癌及裸鼠肝脏原位移植瘤等模型进行抗肿瘤治疗，验证SYK靶向抑制治疗的临床价值。本项目旨在探索胆管癌中SYK促进肿瘤相关巨噬细胞极化的分子机制，及SYK靶向抑制治疗的生物学效应，为胆管癌的靶向治疗提供理论依据。

Tumor-associated macrophages have been shown to correlate with the progression of cholangiocarcinoma (CCA), which have highly malignant and invasive..In our previous study, we found that SYK expression began to up-regulate during the stage of bile duct hyperplasia, and gradually increased with the evolution of CCA. SYK was significantly increased in CCA tissues compared with paratumor tissues, and was positively related with tumor size. On the basis of the Kaplan-Meier survival estimates, the overall survival in CCA patients with high SYK expression is lower in our cohort. The small-molecule inhibitor GS-9973 of SYK can effectively inhibit the tumor growth and infiltration of M2 macrophages in subcutaneous xenografts of nude mice and rat progressive models of CCA..We hypothesized that SYK inhibitor as an effective therapy to attenuate development of cholangiocarcinoma by inhibiting tumor-associated macrophages polarized into M2-phenotype..In this project, the cytological experiment and subcutaneous xenograft in nude mice were used to explore the role of SYK in CCA progression and metastasis and of tumor-associated macrophages phenotype shift induced by SYK. Furthermore, this project makes use of mouse primary hepatocyte isolation, RNA-sequencing, protein mass spectrometric analysis and other technologies to explore the potential mechanism by which SYK-mediation in tumor development via tumor-associated macrophages. The small-molecule inhibitor GS-9973 of SYK was used to treat CCA cells, the subcutaneously transplanted tumors and rat/mouse progressive models of CCA, respectively, to investigate the antitumor efficacy of SYK inhibitors on CCA in vitro and in vivo..The study in this grant proposal will help to elucidate the mechanism of SYK-mediated crosstalk between TAMs and CCA cells and the biological effects of SYK-targeted therapy, which would offer new evidence of targeted therapies for CCA.