Genetic and Molecular Dissection of Wnt Pathway Activation

Wnt 通路激活的遗传和分子剖析

• 批准号: 10417184
• 负责人: Yasmath Ahmed
• 金额: $ 62.98万
• 依托单位: DARTMOUTH COLLEGE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10417184
• 关键词: Address; Adenosine Diphosphate Ribose; Animals; Behavior; Biochemical; Biological; Biological Assay; Cell Differentiation process; Cell Proliferation; Cells; Collaborations; Colorectal Cancer; Complement; Complex; Development; Differentiation and Growth; Disease; Dissection; Drosophila genus; Gene Activation; Genetic; Genetic Transcription; Goals; Homeostasis; Knowledge; Laboratories; Maintenance; Malignant Neoplasms; Mass Spectrum Analysis; Membrane; Modeling; Molecular; Monitor; Natural regeneration; Nuclear; Pathway Analysis; Pathway interactions; Phosphotransferases; Polymerase; Post-Translational Protein Processing; Receptor Activation; Research; Research Personnel; Role; Scaffolding Protein; Signal Transduction; Signal Transduction Pathway; Tankyrase; Tissues; Tumor Suppressor Proteins; WNT Signaling Pathway; Work; beta catenin; cell growth; combat; design; developmental disease; genome wide screen; human disease; in vivo; in vivo Model; innovation; novel therapeutic intervention; programs; receptor; reconstitution; therapeutic target; ubiquitin ligase

PROJECT SUMMARY The evolutionarily-conserved Wnt signal transduction pathway directs cell proliferation and differentiation during animal development and tissue homeostasis. Despite the fact that deregulation of Wnt signaling underlies numerous developmental disorders and cancers, including nearly all colorectal cancers, many of these mechanisms remain poorly understood. Thus, a deeper understanding of the mechanisms that activate this pathway will guide the development of new therapeutic strategies to combat Wnt-driven diseases. The long-term goal of the PI’s research program is to elucidate the mechanisms that activate Wnt signaling during animal development and to use this knowledge to identify control points in the pathway susceptible to therapeutic targeting in Wnt-driven diseases. In support of this effort, the PI and her laboratory group have established innovative in vivo models in Drosophila that have revealed unanticipated functions of three core Wnt pathway effectors: the tumor suppressor, APC; the scaffold protein, Axin; and the ADP-ribose polymerase, Tankyrase. Building on these findings and enabled by genome-wide screens designed to uncover new Wnt signaling regulators, the current project will address three major unsolved questions: 1) how the membrane- associated receptor activation complex is assembled and activated; 2) how the negative regulatory cytosolic beta-catenin destruction complex is inhibited upon Wnt stimulation; and 3) how the nuclear beta-catenin-TCF transcription complex is activated. To elucidate the role of previously unknown Wnt pathway activators, this project will couple genetic, cell biological and biochemical approaches with in vivo assays previously developed by the PI to monitor pathway activation within Wnt signaling gradients. It will apply an innovative focus centered on three components: a deubiquitinating complex essential for Wnt receptor stability, a ubiquitin ligase essential for signaling activation, and a kinase that activates the beta-catenin-TCF transcription complex. This work is complemented by productive collaborations with investigators who have expertise in the biochemical reconstitution of Wnt signaling, Wnt pathway analysis in vertebrate models, and in the identification of Wnt-dependent post-translational protein modifications using mass spectrometry analysis. The successful completion of this work will provide an understanding of: 1) the control of Wnt receptor activity; 2) the control of the beta-catenin destruction complex; 3) the activation of the Wnt transcription complex; and 4) novel therapeutic strategies to target Wnt-driven diseases.

项目概要 进化上保守的Wnt信号转导通路指导细胞增殖和分化 在动物发育和组织稳态过程中。尽管 Wnt 信号传导的放松管制 是许多发育障碍和癌症的基础，包括几乎所有结直肠癌、许多 这些机制仍然知之甚少。因此，对激活机制有更深入的了解 该途径将指导开发新的治疗策略来对抗 Wnt 驱动的疾病。这 PI 研究计划的长期目标是阐明在 动物发育并利用这些知识来识别易受感染途径中的控制点 Wnt 驱动疾病的治疗靶向。为了支持这项工作，PI 和她的实验室小组 在果蝇中建立了创新的体内模型，揭示了三个核心的意想不到的功能 Wnt通路效应子：肿瘤抑制因子，APC；支架蛋白 Axin；和 ADP-核糖聚合酶， 坦聚合酶。以这些发现为基础，并通过旨在发现新 Wnt 的全基因组筛选来实现 对于信号调节器来说，当前的项目将解决三个未解决的主要问题：1）膜如何- 相关受体激活复合物被组装并激活； 2）胞质如何负调节 β-连环蛋白破坏复合物在 Wnt 刺激下受到抑制； 3) 核β-catenin-TCF如何 转录复合物被激活。为了阐明以前未知的 Wnt 通路激活剂的作用， 该项目将把遗传、细胞生物学和生化方法与之前的体内检测结合起来 由 PI 开发，用于监测 Wnt 信号梯度内的通路激活。它将应用创新的 重点集中在三个组成部分：Wnt 受体稳定性所必需的去泛素化复合物、泛素 信号激活所必需的连接酶，以及激活 β-连环蛋白-TCF 转录的激酶 复杂的。这项工作得到了与具有该领域专业知识的研究人员的富有成效的合作的补充。 Wnt 信号的生化重建、脊椎动物模型中的 Wnt 通路分析以及 使用质谱分析鉴定 Wnt 依赖性翻译后蛋白修饰。这 成功完成这项工作将提供以下方面的理解：1）Wnt 受体活性的控制； 2） β-连环蛋白破坏复合物的控制； 3）Wnt转录复合物的激活；和 4) 针对 Wnt 驱动疾病的新治疗策略。