Fam60a-Autophagy通路调控肝再生的作用机制研究

肝脏具有强大的内源性再生能力。然而在许多肝脏疾病中，肝再生过程常常受损，不足以补偿肝细胞乃至整个肝脏的功能损伤。临床上许多肝病治疗依赖于有效的肝脏再生，因此亟需寻找全新的促进肝再生的药物靶点。我们前期基于SCTE-CRISPR高通量筛选技术发现Fam60a是一个调控细胞快速增殖的重要因子；在肝再生模型中Fam60a表达显著上升；Fam60a肝条件性敲除小鼠肝切除后肝再生能力下降；转录组分析表明Fam60a调节肝再生的具体机制可能与自噬有关；Fam60a结合在自噬的重要调节因子Bcl-2的启动子区，并且两者表达存在显著负相关。为进一步揭示Fam60a在肝再生中的调控作用及其机制，本项目采用重组腺病毒、Co-IP等技术，通过原代肝细胞、基因敲除鼠和ALPPS等临床肝脏样品，进一步阐明肝再生中Fam60a调控自噬的具体机制，为今后Fam60a作为促进肝脏再生的潜在药物靶点奠定坚实的理论基础。

Liver has obvious endogenous regeneration ability. However, in many liver diseases, the process of liver regeneration is often damaged, so it is not enough to compensate for the functional damage of hepatocytes and even the whole liver. In clinical practices, many liver diseases depend on effective liver regeneration, so it is urgent to find new drug targets that promote liver regeneration. Based on SCTE-CRISPR high-throughput screening technology, we found that Fam60a is an important factor in regulating rapid proliferation of HepG2 cells; The expression of Fam60a is increased significantly during liver regeneration; The liver regeneration ability is declined after hepatectomy when Fam60a is conditionally knocked out in mice liver; Transcriptome analysis indicates that the mechanism of Fam60a regulating liver regeneration may be related to autophagy; Fam60a binds to the promoter region of Bcl-2, an important regulator of autophagy, and there is a significant negative correlation between the Fam60a and Bcl-2. In order to further clarify the specific mechanism of Fam60a regulating autophagy in liver regeneration, recombinant adenovirus, Co-IP and other technologies will be employed to clarify the function of autophagy in abnormal liver regeneration caused by Fam60a deletion through primary hepatocytes separation and culture, gene knockout mice and clinical liver disease samples. All in all, our research will provide solid references for Fam60a will be one of the potential drug targets of promoting liver regeneration.