通过对耳蜗中Lgr5阳性干细胞转录组的分析，调控促进其再生毛细胞的研究

感音性耳聋主要是由耳蜗毛细胞的不可逆损失造成的。研究表明Lgr5阳性细胞是耳蜗干细胞，通过调节Wnt和Notch信号能促进其增殖分化并再生毛细胞，但耳蜗干细胞的调控机制及其信号通路网络尚不清楚。本项目围绕耳蜗干细胞调控信号通路的研究，比较耳蜗顶圈和底圈Lgr5阳性干细胞增殖分化及毛细胞再生能力的差异，分析其转录组差异并进行信号通路归类，验证并揭示促进耳蜗干细胞增殖分化的信号通路及其之间的协同作用，以期通过信号通路调节能部分恢复听觉功能。拟实现以下研究目标：一、研究耳蜗顶圈和底圈中Lgr5阳性干细胞增殖分化及再生毛细胞能力的差异；二、分析耳蜗顶圈和底圈中Lgr5阳性干细胞转录组的差异，进而分析可能的促进耳蜗干细胞增殖分化的信号通路；三、在离体培养条件下验证这些信号通路的功能，并研究如何通过这些信号通路的协同调控促进毛细胞再生。本项目将为临床上应用耳蜗干细胞治疗听力损失奠定实验基础。

The main reason for sensorineural hearing loss is the irreversible loss of sensory hair cells. Previous studies have reported that Lgr5+ cells are cochlear stem cells and Wnt and Notch signal could induce their proliferation, differentiation and hair cell regeneration. However, the mechanism of cochlear stem cell regulation and the signaling net work are still not clear. This project has been designed to systematically investigate the signaling net work of cochlear stem cell by comparing the ability of hair cell regeneration and the transcriptomes of apex and base Lgr5+ stem cells, with the goal to promote hair cell regeneration from these Lgr5+ cochlear stem cell by regulating these signaling pathway, thus partially recover the hearing function. In this project we propose to fulfill the following aims: Aim1:we will characterize the difference of proliferation, differentiation and hair cell regeneration of Lgr5+ cells in cochlear apex and base; Aim2: we will characterize the transcriptomes of apex and base Lgr5+ cells and analyze the possible signaling pathways that could regulate the cochlear stem cell proliferation and differentiation. Aim3: we will validate the role of these signaling pathways in the in vitro cell culture system, and study the synergetic role between these signaling pathways. The goal of this study is to establish the experimental foundation for the clinical treatment of sensorineural hearing loss by stem cell therapy.