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

基本信息

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
项目类别：
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&apos s Disease Alzheimer&apos s disease brain Alzheimer&apos s disease patient Alzheimer&apos s disease therapeutic Alzheimer&apos 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

项目摘要

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.

项目概要/摘要阿尔茨海默病 (AD) 是美国第六大死因，预计患病率将到 2050 年，这一数字将增加三倍。有效疗法的缺乏促使人们对病理学的分子基础进行研究。 AD 和许多其他神经退行性疾病的特征是病理性原纤维聚集大脑中的 tau 蛋白，以类似朊病毒的方式传播，可能是导致最近，来自人类患者的 tau 原纤维刚性核心的 3.5 Å 结构。 AD 是通过冷冻电子显微镜 (cryo-EM) 解决的。 AD 中 tau 病理学的分子基础，但对蛋白质的其余部分留下了悬而未决的问题：仅 73个残基排列良好，没有关于3R和4R tau掺入的信息目前的提案旨在利用AD tau原纤维研究其结构和动力学。我建议使用 13C、15N 标记的 AD 患者脑源性 tau 纤维进行扩增。这将产生与 AD 脑原纤维结构相同的同位素标记原纤维，我们的小组最近展示了第一个，然后将使用固态核磁共振进行研究。使用 SSNMR 对全长体外 tau 原纤维进行结构和动力学研究。这项工作将开发用于 AD 脑 tau 纤维种子扩增的改进技术，这将为这项工作是对体内淀粉样原纤维进行广泛的生化和结构表征的方法。将利用 Francis Bitter Magnet Lab (FBML) 的可用设施和资源进行麻省理工学院 (MIT) 并将提供以下方面的重要培训机会在我的导师梅教授的指导下进行固态核磁共振和淀粉样蛋白结构生物学 Hong，生物分子固态核磁共振专家，指导过许多成功的科学家完成拟议的学习和培训计划将使我成长为一名独立科学家、导师、沟通者，以追求事业为原则最后，这项工作将为一个主要研究机构的研究人员提供第一个分子见解。 AD tau 原纤维的动态域和其中的亚型混合模式，这应该告知 AD疗法的未来发展。