SIRT6通过IRS2介导的糖酵解调控成牙骨质细胞分化的效应与机制

牙骨质再生是牙周组织再生的金标准。糖酵解作为成骨细胞分化的主要能量来源，在成牙骨质细胞分化中的作用未知。申请人前期发表的论文显示Sirtuin 6（SIRT6）这一重要的糖酵解调节因子抑制了成牙骨质细胞的分化，但其调控机制是否与糖酵解相关有待阐明。为此，本项目首先通过过表达/敲降载体、糖代谢分析等手段探索SIRT6及下游因子胰岛素受体底物2（IRS2）介导的糖酵解在成牙骨质细胞分化中的作用；其次通过染色质免疫共沉淀、双萤光素酶报告基因、表观遗传学分析等技术，进一步研究SIRT6与IRS2的负反馈调控机制；最后应用基因敲除动物模型，体内验证SIRT6通过IRS2介导的糖酵解信号级联在成牙骨质细胞分化过程中的效应与机制。通过本项目的研究，我们拟寻求糖代谢与成牙骨质细胞分化的基因纽带，藉此为了解成牙骨质细胞的代谢与分化的机制提供新的理论基础，也为实现牙骨质再生以及牙周组织再生提供新的思路。

Cementum regeneration is the gold standard for periodontal tissue regeneration. Glycolysis, as the main energy source for osteoblast differentiation, plays an unknown role in cementoblast differentiation. The applicant has been validated that Sirtuin 6(SIRT6), an important glycolytic regulatory factor, inhibited the differentiation of cementoblast, but whether its regulatory mechanism is related to glycolysis remains to be clarified. Hence, we will first explore the role of SIRT6 and downstream factor insulin receptor substrate 2 (IRS2)-mediated glycolysis in cementoblast differentiation through overexpressing/knocking vectors and glucose metabolism analysis; Secondly, we will further detect the negative feedback regulatory mechanism of SIRT6 and IRS2 through chromatin immunoprecipitation, dual-luciferase reporter assay, and epigenetic analysis. Finally, conditional gene knockout animal models will be used to validate the effects and mechanisms of SIRT6 on cementoblast differentiation via IRS2 mediated glycolytic signaling cascade in vivo. Through the implementation of this project, we intend to seek the genetic link between glucose metabolism and cementoblast differentiation, so as to provide a new theoretical basis for understanding the mechanism of metabolism and differentiation of cementoblast, and providing new ideas for the realization of cementum regeneration and periodontal tissue regeneration.