利用带内源标签的小鼠研究SREBP通路调控肝癌发生发展的分子机制

Many studies demonstrate that SREBP (sterol regulatory element-binding protein) pathway is closely associated with cancer development, and hepatocellular carcinoma (HCC) is a high mortality and high incidence worldwide disease. Hence, studying the SREBP pathway in HCC has great significance in cancer prevention and treatment. As our previous study has demonstrated inhibition of SREBP pathway suppresses HCC through repressing inflammation in mice, this project will generate tagging crucial genes in SREBP pathway in androgenetic haploid embryonic stem cells (AG-haESCs) using CRISPR/Cas9 technique. Then establish the SREBP pathway crucial genes knockin mice. Together with different doses of Diethylnitrosamine (DEN) and a chemical compound called betulin blocking SREBP maturation, we will establish HCC mouse models with these knockin mice to mimic cancer developing progress. We will also use tag antibody to analyze the protein components binding to SREBP pathway, and dissect the underlying mechanism to regulate inflammation during carcinoma development. Furthermore, we also plan to focus how the STAT3 (signal transducer and activator of transcription 3) pathway regulates SREBP pathways during this process. We believe, from our proposed study, we will characterize the essential molecular events that SREBP pathway regulates HCC development, which provides a new promising strategy to control HCC and basis for developing new anticancer drugs.

研究表明固醇调节元件结合蛋白(SREBP)调控通路与癌症的发生密切相关，而肝癌是发病率和致死率较高的恶性肿瘤，研究SREBP通路在肝癌发生发展中的分子机制具有重大意义。本申请项目在前期已发表的工作基础上，利用CRISPR/Cas9技术在孤雄单倍体胚胎干细胞上进行原位打靶，快速构建带内源标签的SREBP通路关键基因的敲入小鼠。并运用不同剂量的二乙基亚硝胺(DEN)诱导体内肝癌的发生发展，结合该通路的抑制剂，利用标签抗体研究肝癌发生发展各阶段与SREBP通路关键蛋白相互作用的分子蛋白复合物，全方位系统地揭示SREBP通路参与调控肝癌发生发展中炎症反应的动态分子网络。并重点关注此过程中STAT3信号通路与SREBP通路的互作机制。通过本项目的实施，以期阐明SREBP通路调控肝癌发生发展的重要分子机制，发现新的肿瘤治疗靶点，为开发新型抗癌药物提供重要依据。

肝癌在全球范围内严重影响人们的健康，越来越多的研究证实SREBP代谢调控通路与肝癌的发生发展密切相关，研究SREBP通路在肝癌发生发展中的作用分子机制，对肝癌的预防和治疗具有重大意义。本项目以前期已发表的工作为基础，利用单倍体胚胎干细胞技术，建立了复杂多基因修饰的小鼠模型研究体系，为快速准确构建内源带标签的SREBP通路关键基因的敲入小鼠提供了方法基础。基于基因标签小鼠，通过诱导肝脏肿瘤的发生发展，结合多种分子生物学、组织化学等手段，深入研究SREBP通路影响炎症发生进而影响肝癌发生发展的分子机制，绘制肝癌发生过程中SREBP通路调控炎症反应的动态分子网路。分析临床数据库显示，SREBP通路关键基因在肝癌进展的各个阶段都非常重要，且SREBP通路及其调控分子在人肝癌组织中的表达成特异性、协同性的特点，与肝癌诊断、预后有重要相关性，可以作为肝癌早期诊断与预后的潜在指标。另外，深入研究也揭示了SREBP通路与IL-17炎症途径密切相关，调控机制相互影响，相关的关键分子也可以作为抗癌药物研发的候选靶点。该项目的实施获得的一系列研究结果为肝癌的诊断和开发新的治疗策略提供了依据。