Structure and function of mammalian CPEB2 aggregates in normal and AD brain

正常和 AD 脑中哺乳动物 CPEB2 聚集体的结构和功能

• 批准号: 10539320
• 负责人: Kausik Si
• 金额: $ 45.46万
• 依托单位: STOWERS INSTITUTE FOR MEDICAL RESEARCH
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-15 至 2025-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10539320
• 关键词: Acute; Address; Adult; Alzheimer' s Disease; Amyloid; Amyloid beta-42; Animals; Aplysia; Autopsy; Binding Proteins; Biochemical; Biological; Biological Assay; Brain; CPE-binding protein; Cell physiology; Cells; Clinical Trials; Complex; Cre lox recombination system; Cryoelectron Microscopy; Cytoplasmic Structures; Cytoplasmic Tail; Development; Disease; Drosophila genus; Excision; Failure; Family; Family member; Foundations; Future; Human; Hydrophobicity; Immune; In Vitro; Invertebrates; Knowledge; Length; Light; Link; Lobe; Mammals; Memory; Memory Loss; Memory impairment; Methods; Modeling; Molecular; Mus; Nervous System; Neurodegenerative Disorders; Neurons; Pathologic; Polyribosomes; Process; Property; Prosencephalon; Protein Family; Protein Isoforms; Proteins; Proteome; RANK protein; RNA Binding; RNA-Binding Proteins; Resistance; Resolution; Source; Structure; Synapses; System; TEV protease; Techniques; Tertiary Protein Structure; Testing; Thinking; Tissues; Training; Translations; brain tissue; effective therapy; experience; human subject; hydrophilicity; improved; in vivo; insight; long term memory; mRNA Translation; memory consolidation; memory recall; non-Native; preclinical trial; prion-like; protein aggregation; proteostasis; standard of care; tau Proteins

Project Summary The aggregate failure of pre-clinical and clinical trials in AD has demonstrated that an improved fundamental understanding of memory and how the molecular components of memory are altered in the AD disease process is necessary to develop effective treatment. The broad objective of the project is to identify the biochemical substrates of long-lasting memories in mammals. The current proposal focuses on a family of RNA-binding, cytoplasmic polyadenylation element binding protein (CPEB), that stabilizes memory in invertebrates and mice. Remarkably, CPEB family protein forms non-disease-causing amyloidogenic aggregates and aggregation of CPEB is necessary to stabilize memory. As amyloids are typically linked to disease states, the question remains how similarly structured Aβ42 or Tau proteins can have opposing effects on memory. Therefore, to develop a better understanding of the relationship between amyloids that support memory and amyloids that disrupt memory, we will use a variety of techniques to solve the structure and function of the CPEB family members, CPEB2 and CPEB3, in human and mice. In Aim 1, we will use cryo- electron microscopy to solve the structure of CPEB aggregates from fresh human frontotemporal lobe tissue collected from 25-50-year-old human subjects undergoing tissue removal under the standard of care for their disease. These tissues would have been otherwise discarded. In Aim 2, mice lacking the ability to form aggregates of CPEB2 and CPEB3 will be trained and tested in a one-trial inhibitory avoidance task to assay their ability to form, maintain, and recall memory. In Aim 2 we will also investigate the consequence of CPEB2 and CPEB3 aggregation in translation of mRNA encoding synaptic proteins. The results would be the first to provide direct structural analysis of a functional amyloid linked to memory in mammals, the structural distinctions, if any, between functional and toxic amyloid in the human brain, and precisely link CPEB2 and CPEB3 aggregation and activity to animals’ ability to form or stabilize memory. This knowledge would provide the foundation to investigate in the future how toxic amyloids of Aβ42 or Tau specifically perturb memory.

项目概要 AD 临床前和临床试验的总体失败表明，基础治疗的改善 了解记忆以及记忆的分子成分在 AD 疾病中如何改变 过程对于开发有效的治疗方法是必要的。该项目的总体目标是确定 哺乳动物长期记忆的生化底物。目前的提案主要针对一个家庭 RNA 结合、细胞质多腺苷酸化元件结合蛋白 (CPEB)，可稳定记忆 无脊椎动物和小鼠。值得注意的是，CPEB 家族蛋白形成非致病性淀粉样蛋白 CPEB 的聚合和聚合对于稳定记忆是必要的。由于淀粉样蛋白通常与 疾病状态下，问题仍然是结构相似的 Aβ42 或 Tau 蛋白如何产生相反的作用 靠记忆。因此，为了更好地了解支持淀粉样蛋白之间的关系 记忆和破坏记忆的淀粉样蛋白，我们将使用多种技术来解决结构和 CPEB 家族成员 CPEB2 和 CPEB3 在人和小鼠中的功能。在目标 1 中，我们将使用冷冻 电子显微镜解析新鲜人额颞叶组织 CPEB 聚集体的结构 收集自 25-50 岁的人类受试者，在标准护理下接受组织切除 疾病。否则这些纸巾就会被丢弃。在目标 2 中，小鼠缺乏形成 CPEB2 和 CPEB3 的聚集体将在一项试验抑制性回避任务中进行训练和测试，以进行分析 他们形成、维持和回忆记忆的能力。在目标 2 中，我们还将研究 CPEB2 的后果 编码突触蛋白的 mRNA 翻译中的 CPEB3 聚集。结果将是第一个 提供与哺乳动物记忆相关的功能性淀粉样蛋白的直接结构分析，结构 人脑中功能性淀粉样蛋白和毒性淀粉样蛋白之间的区别（如果有的话），并精确地将 CPEB2 和 CPEB3 聚集和活性对动物形成或稳定记忆的能力的影响。这些知识将提供 为将来研究 Aβ42 或 Tau 的有毒淀粉样蛋白如何特异性扰乱记忆奠定了基础。