CryoEM studies of Wnt3a-FZD8-LRP6 signaling complex in neurogenesis and regeneration

Wnt3a-FZD8-LRP6 信号复合物在神经发生和再生中的冷冻电镜研究

• 批准号: 10798120
• 负责人: Yihe Huang
• 金额: $ 20.31万
• 依托单位: UNIVERSITY OF NEBRASKA LINCOLN
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-15 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10798120
• 关键词: Adult; Alzheimer' s Disease; Biophysics; Brain; Cell membrane; Cells; Communication; Complex; Cryoelectron Microscopy; Degenerative Disorder; Development; Disease; Elements; Event; Foundations; Future; Goals; Homologous Gene; Human; Impairment; Injury; Knowledge; Ligands; Location; Maintenance; Malignant Neoplasms; Maps; Mass Spectrum Analysis; Mediating; Medical; Membrane; Molecular; Natural regeneration; Nebraska; Neurodegenerative Disorders; Neurons; Parkinson Disease; Pathologic; Pathway interactions; Physiological; Play; Proteins; Research; Role; Side; Signal Pathway; Signal Transduction; Signal Transduction Induction; Source; Specificity; Structure; Synaptic plasticity; Therapeutic; Time; Tissues; Visualization; WNT Signaling Pathway; atomic interactions; central nervous system injury; extracellular; neurogenesis; next generation; novel; particle; programs; rational design; receptor; regenerative therapy; stem cell division

Wnt signaling is crucial both for neuronal neurogenesis and for synaptic plasticity and maintenance in the adult brain. Thus, modulating Wnt-mediated neurogenesis and regeneration represents a promising treatment for both central nervous system injuries and neurodegenerative diseases like Parkinson’s and Alzheimer’s. The canonical Wnt3a signaling pathway plays fundamental role in the medically relevant neurogenesis and regeneration and stem-cell renewal. The central event of the Wnt3a pathway is the formation of Wnt3a-FZD8-LRP6 transmembrane complex, which initiates and transduces the signal from the extracellular side into the intracellular side of the cell membrane. The lack of structural information on the intact Wnt3a-FZD8-LRP6 signaling complex or any other complex structure of their homologs greatly hinders our understanding of molecular mechanisms for this fundamental pathway and impairs the development of Wnt-related regenerative therapeutics. The central goal of this research is to determine the structure-based mechanisms of the canonical Wnt3a-FZD8-LRP6 complex by single particle cryo-EM. The structural study will allow us to visualize the atomic interactions between the Wnt3a ligand and its receptors FZD8 and LRP6, map key structural elements that are responsible for signal transduction, and identify potential locations for future therapeutic molecule development. We will also study the structure of the Wnt3a-FZD8-LRP6 complex that purified from native source and investigate the basis of Wnt ligand selectivity and specificity which is a great puzzle in the field considering there are multiple Wnt ligands and receptors. Additionally, we will apply time-resolved cryo-EM studies to resolve the signal transduction dynamics and visualize the Wnt3a signal transduction in near real time. The results of this research program will not only serve as a foundation for understanding the molecular principles by which other Wnt ligands induce signal transduction and related pathological mechanisms in multiple types of diseases including cancer, but also pave the way for the rational design of next-generation therapies for degenerative diseases and injuries.

Wnt信号传导对于神经元的神经发生和突触可塑性以及突触的维持都是至关重要的。 成人大脑因此，调节Wnt介导的神经发生和再生代表了一种有前途的方法。 治疗中枢神经系统损伤和神经退行性疾病，如帕金森氏症， 老年痴呆经典的Wnt 3a信号通路在医学相关疾病中起着重要作用。 神经发生和再生以及干细胞更新。Wnt 3a途径的中心事件是 Wnt 3a-FZD 8-LRP 6跨膜复合物的形成，其启动并转导来自Wnt 3a-FZD 8-LRP 6跨膜复合物的信号。 将细胞外的一侧转移到细胞膜的细胞内一侧。缺乏结构信息， 完整的Wnt 3a-FZD 8-LRP 6信号传导复合物或其同源物的任何其它复合物结构大大 阻碍了我们对这一基本途径的分子机制的理解， Wnt相关再生疗法的发展。本研究的主要目的是确定 通过单颗粒cryo-EM的典型Wnt 3a-FZD 8-LRP 6复合物的基于结构的机制。的 结构研究将使我们能够可视化Wnt 3a配体及其受体之间的原子相互作用 FZD 8和LRP 6，映射负责信号转导的关键结构元件，并鉴定 未来治疗分子开发的潜在位置。我们亦会研究 从天然来源纯化的Wnt 3a-FZD 8-LRP 6复合物，并研究Wnt配体的基础 选择性和特异性，考虑到存在多种Wnt配体， 受体。此外，我们将应用时间分辨冷冻电镜研究来解决信号转导 动力学和可视化Wnt 3a信号转导在接近真实的时间。这项研究成果 该计划不仅可以作为理解其他Wnt分子原理的基础， 配体在多种疾病中诱导信号转导及相关病理机制 包括癌症，但也为下一代治疗的合理设计铺平了道路， 退化性疾病和损伤。