Investigation of Seeded Alzheimer's Disease Tau Fibrils with Solid-State NMR

用固态 NMR 研究种子阿尔茨海默氏病 Tau 原纤维

• 批准号: 10295125
• 负责人: Aurelio James Dregni
• 金额: $ 3.93万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2022-05-26
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10295125
• 关键词: Address; Alzheimer' s Disease; Alzheimer' s disease brain; Alzheimer' s disease patient; Alzheimer' s disease therapeutic; Alzheimer' s disease therapy; Amyloid; Amyloid Fibrils; Biochemical; Brain; Cause of Death; Cryoelectron Microscopy; Cysteine; Development; Disease; Ensure; Environment; Fluorescence; Future; Goals; Heparin; Heterogeneity; Human; In Vitro; Institutes; Institution; Investigation; Isotope Labeling; Label; Left; Length; Massachusetts; Mentors; Mentorship; Methods; Molecular; Molecular Conformation; Mus; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Pathologic; Pathology; Patients; Pattern; Peptide Hydrolases; Predisposition; Prevalence; Progressive Supranuclear Palsy; Property; Protein Isoforms; Proteins; Research; Research Personnel; Resistance; Resources; Role; Sampling; Scientist; Seeds; Sonication; Structure; Techniques; Technology; Testing; Therapeutic; Training; Transmission Electron Microscopy; Trypsin; Work; amyloid structure; beta pleated sheet; career; cofactor; design; effective therapy; experimental study; improved; in vivo; insight; monomer; prion-like; professor; protein aggregation; solid state nuclear magnetic resonance; structural biology; tau Proteins; tau aggregation

PROJECT SUMMARY/ABSTRACT Alzheimer's disease (AD) is the 6th leading cause of death in US, with prevalence expected to triple by 2050. The lack of effective therapies prompts research into the molecular basis of pathology. AD and many other neurodegenerative diseases are characterized by pathological fibril aggregates of the protein tau in the brain, which spread in a prion-like manner and may be the cause of neurodegeneration. Recently, a 3.5 Å structure of the rigid core of tau fibrils from human patients with AD was solved through cryo-electron microscopy (cryo-EM). This structure allowed a glimpse of the molecular basis of tau pathology in AD, but left open questions about the remainder of the protein: only 73 residues were well ordered, and there was no information about the incorporation of 3R and 4R tau isoforms. The current proposal seeks to investigate the structure and dynamics of AD tau fibrils using solid-state NMR. I propose to amplify AD patient brain-derived tau fibrils using 13C, 15N-labelled monomers. This will result in isotopically labelled fibrils with the same structure as the AD brain fibrils, which will then be studied using solid-state NMR. Our group has recently demonstrated the first structural and dynamical investigation of a full-length in vitro tau fibril using SSNMR. If successful, this work will develop improved techniques for seeded amplification of AD brain tau fibrils, which will pave the way for extensive biochemical and structural characterization of in vivo amyloid fibrils. This work will be conducted with the facilities and resources available at the Francis Bitter Magnet Lab (FBML) at the Massachusetts Institute of Technology (MIT) and will provide opportunity for significant training in solid-state NMR and amyloid structural biology under the mentorship of my sponsor, Professor Mei Hong, who is an expert in biomolecular solid-state NMR and has mentored many successful scientists before me. Completion of the proposed studies and training plan will allow me to grow as an independent scientist, a mentor, and a communicator, with the goal of pursuing a career as a principle investigator at a major research institution. Finally, this work will provide the first molecular insights into the dynamic domains of AD tau fibrils and the isoform mixing pattern within them, which should inform future development of AD therapies.

项目总结/摘要 阿尔茨海默病（AD）是美国第六大死亡原因，预计患病率将 到2050年翻三倍缺乏有效的治疗方法促使人们研究病理学的分子基础。 AD和许多其他神经退行性疾病的特征在于， 大脑中的tau蛋白，以朊病毒样的方式传播，可能是导致 神经变性最近，来自人类患者的tau原纤维的刚性核心的3.5 μ m结构， 通过冷冻电子显微镜（cryo-EM）解决AD。这个结构让我们得以一瞥 AD中tau病理学的分子基础，但留下了关于蛋白质其余部分的开放性问题：仅 73个残基排列良好，没有关于3R和4 R tau掺入的信息 同种型。目前的建议旨在研究AD tau纤维的结构和动力学， 固态NMR。我建议使用13 C、15 N标记来扩增AD患者脑源性tau纤维 单体。这将产生与AD脑原纤维具有相同结构的同位素标记的原纤维， 然后用固态核磁共振进行研究。我们的团队最近展示了第一个 使用SSNMR对全长体外tau原纤维结构和动力学研究。如果成功，这 这项工作将开发用于AD脑tau纤维的种子扩增的改进技术，这将为AD的研究铺平道路。 对体内淀粉样纤维进行广泛的生物化学和结构表征的方法。这项工作 将在弗朗西斯苦磁铁实验室（FBML）的设施和资源下进行， 马萨诸塞州理工学院（MIT），并将提供以下方面的重要培训机会 固态核磁共振和淀粉样蛋白结构生物学的指导下，我的赞助商，梅教授 Hong是生物分子固态核磁共振的专家，并指导了许多成功的科学家 在我之前。完成拟议的学习和培训计划将使我能够成长为一名 独立的科学家，导师和沟通者，以追求事业为原则 一家大型研究机构的研究员。最后，这项工作将提供第一个分子的见解， AD tau原纤维的动态结构域和其中的同种型混合模式，这应该告知 AD治疗的未来发展。