肝内胆管癌（Intrahepatic Cholangiocarcinoma,ICC）是人类预后最差的肿瘤之一，由于化疗耐药等原因，中位生存期不足12个月，其发生及发展机制仍不明确。本课题预实验发现5-hmC及其催化酶TET1在ICC中显著上调与患者的预后密切相关，敲减TET1能抑制ICC进展。我们拟在ICC细胞株及肿瘤组织中进行ChIP-seq等表观基因组学实验，挖掘以TET1为核心的ICC表观基因组改变；另外，预实验发现抑制染色质中的RNA能显著下调TET1在染色质中的定位，将进行CLIP-seq及系列功能学实验明确LncRNA在TET1定位中的重要作用；此外，预实验发现TET1相关的染色质三维结构在ICC和正常胆管内皮细胞之间存在显著差异，将进行ChIA-PET等实验阐明TET1相关的三维染色质重构与ICC的关系。本课题将揭示ICC的发生及发展的全新机制，并挖掘新的标记物及治疗靶点。

Intrahepatic Cholangiocarcinoma (ICC) is a common malignancy with poor prognosis (Median survival time: 12 months) due to the Chemoresistance, etc. Currently, the mechanisms of ICC pathogenesis were poorly understood. Our preliminary data showed that the amount of 5-hydromethyl cytocine (5-hmC), a critical intermediate of DNA methylation, was significantly upregulated in the ICC tissues from the patients as compared to normal paratomor control. Moreover, Ten-Eleven Translocation 1(TET1) protein, a critical methylcytosine dioxygenase to catalyze the formation of 5-hmC, was increased concomitantly. Intriguingly, we demonstrated that the TET1 protein expression was significantly correlated with overall survival time of ICC patients. Moreover, our in vitro and in vivo functional studies unraveled that knockdown of TET1 dramatically inhibited the growth and proliferation of ICC, indicating the potential role of TET1 in the pathogenesis and development of ICC. Futhermore, by ChIA-PET approach, we found the great disparity in terms of TET1 assoicated 3D chromatin structure in ICC as comparted to normal cholangiocyte, highlighting the potential TET1 associated 3D chromatin structure in the gene regulation of ICC. Based on these exciting preliminary data, we aim to 1) unravel the uniqueness of TET1 assoicated epigenome in ICC by ChIP-seq, hMeDIP-seq and RNA-seq; 2) to demystify the role of LncRNA in guiding the genomic localization of TET1 in ICC by CLIP-seq and other approaches; 3) to decode the TET1 associated 3D chromatin structure and its influence by LncRNA in gene regulation of ICC by integration of ChIA-PET and aforementioned ChIP-seq, RNA-seq dataset. Taken together, our innovative project will disclose the complete new mechanism of the tumorigenesis and progression of ICC, and uncover novel biomarkers and potential therapeutic targets.

肝内胆管癌（Intrahepatic Cholangiocarcinoma,ICC）是人类预后最差的肿瘤之一，由于化疗耐药等原因，中位生存期不足12个月，其发生及发展机制仍不明确。本课题通过完成ICC细胞株和ICC组织的ChIP-seq、CLIP-seq、ChIA-PET等实验，发现5-hmC及其催化酶TET1在ICC中显著上调与患者的预后密切相关，敲减TET1能抑制ICC进展。我们在ICC细胞株及肿瘤组织中进行ChIP-seq等表观基因组学实验，进一步挖掘以TET1为核心的ICC表观基因组改变；另外，实验发现抑制染色质中的RNA能显著下调TET1在染色质中的定位，进行了CLIP-seq及系列功能学实验。通过上述实验将进行生物信息学分析，前期分析揭示抑制ICC的新生RNA的产生能下调TET1在ATCB基因上的结合，对ICC后续的研究具有重要的研究意义。