Synaptic mechanisms of tau-mediated pathogenesis in human iPSC-derived neurons

人 iPSC 衍生神经元中 tau 介导的发病机制的突触机制

• 批准号: 10092879
• 负责人: Tara Tracy
• 金额: $ 12.44万
• 依托单位: BUCK INSTITUTE FOR RESEARCH ON AGING
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-04-15 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10092879
• 关键词: Advisory Committees; Affect; Alzheimer' s Disease; Behavioral; Binding; Biotin; Brain; CRISPR/Cas technology; California; Career Mobility; Cell Line; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Cognitive; Cognitive deficits; Committee Members; Communities; Country; Disease; Disease model; Elderly; Electrophysiology (science); Engineering; Equilibrium; Event; Frontotemporal Lobar Degenerations; Functional disorder; Funding; Genetic Engineering; Glutamate Receptor; Glutamates; Goals; Human; Impaired cognition; Impairment; Individual; Induced pluripotent stem cell derived neurons; Inherited; Institutes; Knock-out; Label; Lead; Leadership; Learning; Link; Mass Spectrum Analysis; Mediating; Memory Loss; Memory impairment; Mentors; Microtubule-Associated Proteins; Modeling; Molecular; Mus; Mutation; Nerve Degeneration; Neurodegenerative Disorders; Neuronal Dysfunction; Neurons; Pathogenesis; Pathogenicity; Patients; Physiological; Process; Professional Competence; Property; Proteins; Proteomics; Public Health; Research; Research Personnel; Research Proposals; Resources; San Francisco; Scientist; Signal Transduction; Signaling Molecule; Societies; Synapses; Synaptic Transmission; Tauopathies; Techniques; Testing; Therapeutic Intervention; Toxic effect; Training; Transgenic Mice; Transplantation; Universities; age related; age related neurodegeneration; aging demographic; base; career; efficacy evaluation; genome editing; human disease; in vivo; induced pluripotent stem cell; innovation; insight; novel strategies; novel therapeutic intervention; overexpression; postsynaptic; presynaptic; progressive neurodegeneration; skills; stem cell differentiation; synaptic function; targeted treatment; tau Proteins; tau aggregation; tau dysfunction; tau mutation; therapy outcome; transcription factor; transmission process

PROJECT SUMMARY/ABSTRACT Tauopathies are currently the most prevalent neurodegenerative diseases in our country and represent a public health crisis for our rapidly aging demographic. People living with tauopathy, including Alzheimer's disease (AD) and frontotemporal lobar degeneration with tau inclusions (FTLD-tau), are afflicted with severe and devastating memory loss. There are few treatments available for people suffering from cognitive decline and there is no cure. Memory impairments in AD are highly correlated with synapse loss in the brain, suggesting that synaptic function is particularly vulnerable and may be the most promising target for a successful therapeutic outcome. However, the pathogenic events that lead to synapse decline in tauopathy are not well understood. The accumulation of tau, a microtubule-associated protein, in the brain is a hallmark of tauopathy that coincides with progressive neurodegeneration. How tau contributes to synaptic decline is unclear. The main objectives of the proposed research are to determine how tau affects synaptic function in human induced pluripotent stem cell (iPSC)- derived neurons and to identify tau-dependent mechanisms that modulate synapses. This research will apply cutting-edge techniques including targeted genome editing of iPSCs and NGN2-induced iPSC differentiation into neurons. CRISPR-based genome editing of tauopathy patient derived iPSCs with FTLD-tau mutations will generate isogenic iPSCs with corrected mutations. A wild-type tau iPSC line will also be genetically modified using CRISPR to generate isogenic iPSCs carrying mutations that cause familial FTLD-tau. These iPSC lines will be used to determine how tau affects synaptic function in human neuronal models of FTLD-tau (Aim 1). APEX2-based proximity-dependent biotin labeling and quantitative mass spectrometry analyses will be used to identify tau-dependent mechanisms that promote synaptic dysfunction (Aim 2). Finally, tauopathy patient iPSC- derived neurons will be transplanted into mouse brain to determine how tau alters excitatory and inhibitory synaptic transmission in vivo (Aim 3). Further scientific training will enable the candidate to delineate mechanisms linking pathogenic tau and synaptic toxicity in a human disease model. A team of co-mentors and advisory committee members will provide guidance and support for the candidate's research proposal and career advancement. The vibrant scientific community and abundant resources at the Gladstone Institutes and the University of California, San Francisco will enhance the candidate's training. The short-term goals of the candidate are to 1) acquire additional techniques to investigate tauopathy using human neuronal models 2) acquire leadership, mentoring and networking skills for career advancement 3) obtain funding for research as a junior investigator. The long-term goals of the candidate are to make major contributions in tauopathy research by uncovering how tau triggers pathogenic events that lead to neuronal dysfunction and to establish novel strategies for recovering synaptic function in the brain to treat cognitive decline.

项目总结/摘要 Tauopathies是目前我国最流行的神经退行性疾病， 我们快速老龄化人口的健康危机。患有tau蛋白病的人，包括阿尔茨海默病（AD） 和额颞叶变性与tau夹杂物（FTLD-tau），患有严重的和毁灭性的 失忆对于患有认知能力下降的人来说，几乎没有治疗方法，也没有治愈方法。 AD患者的记忆障碍与大脑中的突触丢失高度相关，这表明突触功能 是特别脆弱的，并且可能是成功治疗结果的最有希望的靶点。然而，在这方面， 导致Tau蛋白病中突触下降的致病事件还没有被很好地理解。的积累 tau蛋白是一种微管相关蛋白，在大脑中是tau蛋白病的标志，与进行性tau蛋白病一致。 神经变性tau蛋白如何导致突触衰退尚不清楚。建议的主要目标 研究旨在确定tau蛋白如何影响人类诱导多能干细胞（iPSC）的突触功能- 衍生的神经元，并确定调节突触的tau依赖性机制。这项研究将适用于 尖端技术，包括iPSC的靶向基因组编辑和NGN 2诱导的iPSC分化为 神经元具有FTLD-tau突变的tau病变患者衍生的iPSC的基于CRISPR的基因组编辑将 产生具有校正突变的同基因iPSC。野生型tau iPSC系也将被遗传修饰 使用CRISPR产生携带引起家族性FTLD-tau的突变的同基因iPSC。这些iPSC细胞系 将用于确定tau如何影响FTLD-tau的人神经元模型中的突触功能（Aim 1）。 将使用基于APEX 2的邻近依赖性生物素标记和定量质谱分析， 鉴定促进突触功能障碍的tau依赖性机制（目的2）。最后，tau蛋白病患者iPSC- 将衍生的神经元移植到小鼠脑中以确定tau如何改变兴奋性和抑制性 体内突触传递（目的3）。进一步的科学训练将使候选人能够描绘出 在人类疾病模型中连接致病性tau和突触毒性的机制。一组共同导师和 顾问委员会成员将为候选人的研究计划和职业生涯提供指导和支持 进步。充满活力的科学界和丰富的资源，在格莱斯顿研究所和 加州大学旧金山弗朗西斯科将加强对应聘者的培训。短期目标 候选者是1）获得使用人类神经元模型研究tau蛋白病的额外技术2） 获得领导力，指导和网络技能，以促进职业发展3）获得研究资金， 初级调查员候选人的长期目标是在tau蛋白病研究中做出重大贡献 通过揭示tau蛋白如何触发导致神经元功能障碍的致病事件， 恢复大脑中突触功能以治疗认知能力下降的策略。