RNA去甲基化酶FTO介导的lncRNA-m6A修饰对肝癌细胞代谢重编程的调控作用及机制研究

Recently, the energy metabolism reprogram is recognized as one of the hallmarks of cancer cells.Therefore, investigating the molecular mechanism of HCC energy metabolism reprograms has important theoretical guidance and clinical significance. It has been reported that N6-Methyladenosine (m6A) modification has been implicated in many biological processes and also plays an important role in tumor progression. In this research, we demonstrated that m6A modifications are decreased in HCC patients of metabolic disorders and that FTO (Alpha-Ketoglutarate Dependent Dioxygenase) is the main factor involved in aberrant m6A modification. We also found that FTO is highly associated with cell proliferation and energy metabolism reprogram of HCC. On the other side, long noncoding RNA (lncRNA) is proved to be associated with tumor energy metabolism reprogram and m6A modification. In the proposal, we aim to perform a high-throughput screening (RNA-seq and MeRIP-seq) to screen the m6A modified lncRNAs which is implicated in HCC energy metabolism reprogram. Next, we will construct the regulation network of FTO-m6A-lncRNAs in HCC energy metabolism reprogram and further investigate the detailed molecular mechanism of the FTO-m6A-lncRNAs in regulating HCC cell metabolism and hepatic carcinogenesis. In addition, we will examine the expression level of the FTO and candidate lncRNAs in a large sample of HCC patient liver tissues to determine the correlation between candidate lncRNAs expression and energy metabolism, prognosis, recurrence and survival of HCC patients. Finally, we will design and synthesize the inhibitors of FTO and it will benefit for the discovery of novel therapeutic targets for the treatment of HCC. Together, we aim to investigate the function and mechanism of FTO mediated lncRNA m6A modification in HCC energy metabolism reprogram and offer possible targets for HCC therapy and prediction.

目前肿瘤细胞能量代谢重编程已公认是肿瘤细胞的十大特征之一，因此探讨肝癌能量代谢重编程的分子机理，具有重要的理论指导意义和临床价值。有研究表明RNAm6A修饰在肿瘤进展中发挥重要作用，但其在肝癌细胞代谢重编程中的机制尚不清楚。本项目前期研究发现RNA去甲基化酶FTO是代谢异常肝癌中m6A修饰紊乱的关键调控基因，其介导的m6A修饰可促进肝癌细胞增殖能力及代谢水平，另lncRNA也参与调控了肝癌细胞代谢重编程且受到m6A修饰。因此本项目拟利用MeRIP-seq技术筛选肝癌细胞代谢重编程中受到m6A调控的lncRNA，构建FTO-m6A-lncRNA的肝癌细胞代谢重编程的调控网络，并阐明其在肝癌细胞代谢调节中的具体分子机制。同时本项目将在大规模肝癌样本中检测FTO及核心lncRNA的表达水平，分析其与肝癌患者预后及代谢等临床病理特征之间的相关性，并将设计合成FTO抑制剂，为肝癌的诊断治疗提供参考。

目前肿瘤细胞能量代谢重编程已公认是肿瘤细胞的十大特征之一，因此探讨肝癌能量代谢重编程的分子机理，具有重要的理论指导意义和临床价值。有研究表明m6A修饰在肿瘤进展中发挥重要作用，但其在肝癌细胞代谢重编程中的机制尚不清楚。本项目利用MeRIP-seq及RNA-seq等高通量测序技术、生物信息学分析、分子生物学等方法筛选得到FTO介导m6A修饰的lncRNA-MDAL，并且利用实验证实lncRNA-MDAL参与调控了肝癌细胞糖类及脂类代谢的过程且受到RNA m6A修饰。下一步将构建FTO-m6A-lncRNA-MDAL的肝癌细胞代谢重编程的调控网络，并揭示其在肝癌细胞代谢调节中的具体分子机理:lncRNA-MDAL参与调控了肝癌糖类代谢过程且受到m6A修饰。并构建了FTO-m6A-lncRNA-MDAL的肝癌细胞糖类代谢的调控网络，通过实验证实LncRNA-MDAL可与SAF-A蛋白结合调控了Pentose Phosphate Pathway促进了肝癌细胞糖类代谢水平。同时本项目在大样本肝癌组织中检测了FTO及核心lncRNA-MDAL的表达水平，结合临床资料分析证实其与肝癌代谢、预后等临床指标相关。并进一步设计合成了FTO抑制剂，检测其对肝癌细胞代谢的影响及其对肝癌的治疗作用。