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.

项目摘要 进化保存的Wnt信号转导途径指导细胞增殖和分化 在动物发育和组织稳态期间。尽管Wnt信号的放松管制 基础众多发育障碍和癌症，包括几乎所有有色癌症，许多 这些机制仍然知之甚少。这是对激活机制的更深入的理解 该途径将指导开发新的治疗策略，以打击WNT驱动的疾病。这 PI研究计划的长期目标是阐明激活Wnt信号传导的机制 动物发展并利用这些知识来识别容易受到影响的路径中的控制点 WNT驱动疾病中的治疗靶向。为了支持这项努力，PI和她的实验室小组有 果蝇中建立的创新体内模型，这些模型揭示了三个核心的意外功能 Wnt途径效应：肿瘤抑制剂，APC；支架蛋白Axin；和ADP-核糖聚合酶， 坦克酶。以这些发现为基础，并由全基因组屏幕启用，旨在发现新的Wnt 信号调节器，当前项目将解决三个主要未解决的问题：1）膜如何 相关的接收器激活复合物被组装并激活； 2）负调控性胞质如何 Wnt刺激抑制了β-catenin的破坏复合物； 3）核β-catenin-TCF如何 转录复合物被激活。为了阐明先前未知的Wnt途径激活剂的作用， 项目将将遗传性，细胞生物学和生化方法与之前的体内分析相结合 由PI开发以监视Wnt信号传导梯度内的途径激活。它将应用创新 重点集中于三个组成部分：泛素化复合物对于Wnt接收器稳定性至关重要，泛素 连接酶对于信号激活必不可少的，以及激活β-catenin-TCF转录的激酶 复杂的。这项工作是由与具有专业知识的调查人员合作完成的 Wnt信号传导的生化重构，脊椎动物模型中的WNT途径分析以及 使用质谱分析鉴定WNT依赖性后翻译后蛋白质修饰。这 这项工作的成功完成将提供：1）Wnt受体活动的控制； 2） β-catenin破坏复合物的控制； 3）Wnt转录复合物的激活；和4） 针对WNT驱动疾病的新型治疗策略。