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

项目摘要/摘要 阿尔茨海默病(AD)是美国第六大死亡原因，患病率预计将 到2050年翻两番。缺乏有效的治疗方法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的纤维聚集 脑组织中的tau蛋白，它以类似普里恩的方式传播，可能是 神经退行性变。最近，来自人类患者的tau纤维的硬核的3.5？结构 用冷冻电子显微镜(Cryo-EM)检测AD。这一结构使人们得以一窥 阿尔茨海默病中tau病理的分子基础，但关于蛋白质的其余部分仍未解决问题：仅 73个残基排列有序，没有关于3R和4R tau掺入的信息 异构体。目前的建议试图研究AD tau纤维的结构和动力学 固态核磁共振。我建议用13C，15N标记扩增AD患者脑源性tau纤维 单体。这将导致同位素标记的纤维具有与AD脑纤维相同的结构， 然后将使用固态核磁共振进行研究。我们的团队最近展示了第一个 用SSNMR研究体外全长tau纤维的结构和动力学。如果成功，这将是 工作将开发用于AD脑tau纤维种子放大的改进技术，这将为 体内淀粉样纤维的广泛生化和结构表征方法。这部作品 将利用弗朗西斯苦磁实验室(FBML)现有的设施和资源进行，地址为 麻省理工学院(MIT)将提供重要的培训机会 在我的赞助人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 洪是生物分子固态核磁共振方面的专家，曾指导过许多成功的科学家 在我之前。完成拟议的学习和培训计划将使我成为一名 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调查员。最后，这项工作将提供第一个分子洞察 AD tau纤维的动态域及其内部的异构体混合模式 阿尔茨海默病治疗的未来发展。

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.