TGFβ-STAT3信号轴通过调节中性粒细胞表型和功能影响肝豆状核变性疾病进展的作用及机理研究

肝豆状核变性是我国高发的遗传代谢性疾病之一，然而，目前关于该疾病如何进展的分子和细胞调控机理研究不足。中性粒细胞表型和功能是否影响该疾病进展目前还没有报道。申请人前期通过活体成像、细胞轨迹追踪、流式分选及外源添加小分子抑制剂等斑马鱼模型的实验，充分提示了中性粒细胞表型和功能对该疾病进展的重要性，且TGFβ可能参与调控该过程。数据库比对分析发现，STAT3是中性粒细胞表型和功能改变的必需因子。依据前期基础及文献分析，我们提出TGFβ-STAT3信号轴通过调节中性粒细胞表型和功能促进肝豆状核变性疾病进展的假说。本项目拟从小鼠模型入手，结合条件敲除转基因鱼和双敲除小鼠的使用，以及骨髓中性粒细胞与模型细胞株共培养的体外实验，采用流式分析、免疫印迹、活体成像、细胞轨迹追踪、siRNA干扰和荧光素酶报告分析等多种技术手段，从现象、功能和机理三个层次阐明研究假说。本研究将为肝豆状核变性的治疗提供新靶点。

Wilson's disease (OMIM#277900) is one of the most common inherited metabolic diseases. The mechanism of its progression has not been clarified. Whether the phenotype and function of neutrophils influence the progression of Wilson's disease has not been reported. Our previous studies in zebrafish models suggested that the phenotype and function of neutrophils changed during development of the disease, and TGFβ was involved in this process. STAT3 is an essential factor for the change of neutrophil phenotype and function based on the comparison of microarray data from neutrophil subtypes (GSE101584) and transcription factor database (TF checkpoint database). Based on our previous experiments and literature review, we hypothesize that TGFβ-STAT3 signaling axis accelerates the development of Wilson's disease by regulating neutrophil phenotype and function. To confirm this, our project will first clarify the neutrophil phenotype and function in the progression of mouse models. Secondly, the roles of neutrophil phenotype and function in the progression of the disease will elucidate in conditional knockout transgenic fish and double knockout mice. Finally, the molecular mechanism of TGFβ-STAT3 signaling axis in regulating the phenotype and function of neutrophils will decipher through the in vivo transfusion of small molecular compounds in mice and co-culture of isolated bone marrow neutrophils with ATP7B-/- HepG2. This study will provide a new target for the treatment of Wilson's disease.