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: 10328953
• 负责人: Kausik Si
• 金额: $ 45.19万
• 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/10328953
• 关键词: Acute; Address; Adult; Alzheimer' s Disease; Amyloid; Amyloid beta-42; Animals; Aplysia; Autopsy; 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; Genetic Translation; Human; Hydrophobicity; Immune; In Vitro; Invertebrates; Knowledge; Length; Light; Link; Lobe; Mammals; Memory; Memory Loss; Memory impairment; Methods; Modeling; Molecular; Mus; Nervous system structure; 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; base; brain tissue; effective therapy; experience; human subject; hydrophilicity; improved; in vivo; insight; long term memory; 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.

项目摘要 AD前临床和临床试验的总失败表明，基本的基本性改善 了解记忆以及在AD病中如何改变记忆的分子成分 过程对于开发有效的治疗是必要的。该项目的广泛目标是确定 哺乳动物长期记忆的生化底物。当前的提议着重于一个家庭 RNA结合，细胞质聚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家族蛋白形成非疾病的淀粉样蛋白蛋白 CPEB的聚合和聚集是稳定内存的必要条件。由于淀粉样蛋白通常链接到 疾病状态下，问题仍然是结构化的Aβ42或tau蛋白的类似结构如何具有相反的作用 在内存。因此，要更好地了解支持的淀粉样蛋白之间的关系 内存和淀粉样蛋白会破坏内存，我们将使用各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AIM 1中，我们将使用冷冻 电子显微镜解决了新鲜人额叶叶组织的CPEB聚集体的结构 从25-50岁的人类受试者根据其护理标准进行组织去除的受试者 疾病。这些组织本来可以被丢弃。在AIM 2中，老鼠缺乏形成能力 CPEB2和CPEB3的聚集体将在一次性抑制避免任务中进行培训和测试以测定 他们形成，维护和回忆记忆的能力。在AIM 2中，我们还将调查CPEB2的后果 和CPEB3在MRNA编码突触蛋白的翻译中的聚集。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 在人脑中的功能和有毒淀粉样蛋白之间的区别（如果有的话），并精确地将CPEB2和 CPEB3的聚集和活动对动物形成或稳定记忆的能力。这些知识将提供 未来研究Aβ42或TAU的淀粉样蛋白如何特别扰动记忆的基础。