“间质-淋巴内皮转化”在肝内胆管癌淋巴管新生中的作用机制研究

Lymph node metastasis of intrahepatic cholangiocarcinoma (ICC) represents one of the typical characteristics different from hepatocellular carcinoma, and remains the vital risk factor which badly affects patients’ prognosis as well. However, the molecular mechanism of lymph node metastasis remains unclear. The applicant previously found that some lymphatic endothelium-like cancer associated fibroblasts (double staining for PDPN and α-SMA) in the tumor microenvironment of ICC are highly associated with poor prognosis and ICC lymphangiogenesis, which indicates cancer associated fibroblasts (CAFs) may be likely adopt lymphatic endothelium phenotype by mesenchymal-to-lymphatic endothelial transition. Taken together, we here plan to (1) identify the potential role of PDPN(+) CAFs for mesenchymal-to-lymphatic endothelial transition in tumor microenviroment; (2) verify the contribution of PDPN(+) CAFs for ICC metastasis; (3) clarify the underlying mechanism for the formation of PDPN(+) CAFs by integrating NGS and cas-9 technologies; (4) prevent metastasis of ICCs by targeting PDPN(+) CAFs in vivo. If successful, it will deepen our understanding of the molecular mechanism of ICC lymph node metastasis, which could provide a new strategy for the future targeted microenvironment intervention.

淋巴结转移是肝内胆管癌（ICC）区别于肝细胞癌的典型特征，也是影响其预后的关键因素，然而调控ICC淋巴结转移的分子机理尚不明确。申请人前期发现ICC肿瘤微环境中存在一群淋巴内皮样癌相关成纤维细胞（淋巴内皮细胞特异标志物PDPN和α-SMA双阳性），该群细胞数目与ICC患者预后不良（特别是淋巴管新生）密切相关，提示癌相关成纤维细胞（CAF）有可能通过间质-内皮转化获得淋巴内皮细胞表型从而诱导淋巴管形成促进ICC淋巴结转移。基于此本研究计划首先明确PDPN+ CAF在ICC肿瘤微环境“间质-淋巴管内皮转化”中的关键生物学功能；其次结合共培养模型分析该群细胞在ICC转移中的核心作用；并应用高通量测序、cas-9等技术阐明该群细胞转化的分子机理；最后结合在体模型探讨干预PDPN+ CAF细胞形成阻断肿瘤转移。项目的实施将加深我们对ICC淋巴结转移分子机理的认识，为今后靶向微环境治疗肿瘤提供新策略。

淋巴结转移是肝内胆管癌（ICC）区别于肝细胞癌的典型特征，也是影响其预后的关键因素，然而调控ICC淋巴结转移的分子机理尚不明确。申请人前期发现特异性癌相关成纤维细胞有可能通过分泌促淋巴管细胞因子从而诱导淋巴管形成促进ICC淋巴结转移。基于此本研究首先明确FGF2-FGFR1通路在调控淋巴管生成中的关键生物学功能；其次发现VEGFC-VEGFR3通过调控HK2表达影响淋巴管新中的核心作用；并应用双荧光素酶报告基因实验、cas-9等技术阐明该群细胞转化的分子机理；最后结合在体模型探讨干预FGF通路及VEGF通路阻断HK2表达从而抑制淋巴管新生。项目的实施将加深我们对ICC淋巴结转移分子机理的认识，为今后靶向微环境治疗肿瘤提供新策略。