基于化学遗传学策略研究NDR1激酶抑制剂对细胞命运的影响

Human NDR1 (a nuclear serine/threonine kinase) is a member of evolutionary conserved NDR family kinases, which play important roles in many biological processes, such as function in mitotic regulation. Our further investigations revealed that PLK1-mediated phosphorylation suppresses the activity of NDR1 to ensure proper spindle orientation and signaling networks. Importantly, knockdown of NDR1 perturbs spindle orientation. However, the precise functions and the underlying regulatory mechanisms remain unclear. So, discovering potent and selective small-molecule inhibitors of the NDR1 has become one of the most active fields. In this project, we will explore a virtual screening-based inhibitor approach targeting NDR1 kinase and identify phosphorylation substrates of NDR1 by chemical genetics to investigate the molecular mechanisms of spindle orientation in vitro and in vivo. This project will provide novel insights into a better understanding of PLK1-NDR1 kinase cascade in mitotic spindle orientation dynamics and plasticity with bioinformatics and chemical probe methods.

人类NDR激酶家族非常保守，NDR1激酶参与有丝分裂相关调控等细胞生理过程。我们最新研究揭示: PLK1磷酸化修饰NDR1并动态调控其活性从而保证正确的纺锤体定向。为了进一步解析NDR1调控纺锤体正确定向的分子机制与信号转导网络, 本项目将基于虚拟筛选开发NDR1的小分子抑制剂并利用NDR1小分子抑制剂为探针解析纺锤体定向机制。以化学小分子探针为手段系统阐明细胞可塑性调控所涉及的信号转导分子机理对于理解生命活动的本质具有十分重要的意义。NDR1激酶化学探针的发掘将使我们能够阐明NDR1激酶活性在有丝分裂期的时空动力学特征及其调控染色体分离保真性的分子机制。

人类NDR激酶家族非常保守，NDR1激酶参与有丝分裂相关调控等细胞生理过程。我们最新研究揭示: PLK1磷酸化修饰NDR1并动态调控其活性从而保证正确的纺锤体定向。为了进一步解析NDR1调控纺锤体正确定向的分子机制与信号转导网络, 本项目将基于虚拟筛选开发NDR1的小分子抑制剂并利用NDR1小分子抑制剂为探针，解析纺锤体定向机制。本项目已发掘了NDR1化学小分子探针LCD1825，系统阐明了细胞可塑性调控所涉及的信号转导通络，从纳米尺度解析有丝分裂纺锤体正确定向的调控机制。LCD1825通过干扰NDR1的底物CENP-T的磷酸化修饰，导致其无法正确定位，产生滞后染色体。因此，NDR1相互作用蛋白质群的磷酸化、乙酰化等翻译后修饰关系的建立，对于解析有丝分裂时空动态性调控具有十分重要的意义。基于生物信息分析和化学生物学方法的应用，NDR1激酶化学探针LCD1825的发掘，使我们能够阐明SET7/9-PLK1-NDR1信号轴在有丝分裂期的时空动力学特征及其调控纺锤体定向的动态可塑性。