Yeast as a gateway to conquering protein misfolding diseases.

酵母是征服蛋白质错误折叠疾病的门户。

• 批准号: 10396270
• 负责人: SUSAN W LIEBMAN
• 金额: $ 14.78万
• 依托单位: UNIVERSITY OF NEVADA RENO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10396270
• 关键词: Address; Affect; Alzheimer' s Disease; Amyloid; Amyotrophic Lateral Sclerosis; Area; Binding Proteins; Cell physiology; Cells; Characteristics; Cytoplasmic Granules; Dementia; Development; Disease; Funding; Gene-Modified; Goals; Growth; Human; Learning; Liquid substance; Methods; Modeling; Mus; Mutation; Neurodegenerative Disorders; Neurons; Parkinson Disease; PrP; Prions; Proteins; Risk Factors; TDP-43 aggregation; Therapeutic; Titrations; Toxic effect; Variant; Work; Yeast Model System; Yeasts; cellular targeting; disorder risk; fly; frontotemporal lobar dementia-amyotrophic lateral sclerosis; gain of function; human disease; insight; overexpression; parent grant; prion seeds; prion-like; protein TDP-43; protein aggregation; protein misfolding; therapeutic target; tool; yeast prion

Abstract of the Funded Parent Grant. Certain proteins misfold to form self-seeding prion-like aggregates associated with disease. We focus on one such protein, TDP-43, because it is the major protein associated neuronal aggregates in several neurodegenerative diseases including amyotrophic lateral sclerosis (ALS), frontotemporal dementia and LATE. LATE is a recently described prevalent TDP-43 proteinopathy that causes dementia that is often misdiagnosed as Alzheimer’s disease (AD). In addition, TDP-43 is found in aggregates associated with AD and Parkinson’s. Since TDP-43 forms aggregates and is toxic (inhibits growth) in yeast, a powerful approach to find therapeutic targets has been to identify yeast genes that modify TDP-43 toxicity. The relevance of the yeast model to human disease is clear because several yeast genes that modify toxicity of human misfolding disease proteins, including TDP-43, are homologs of new or known human disease risk factors. We will continue to study the genesis and toxicity of TDP-43 aggregates in yeast building on our expertise with yeast self-seeding prion proteins. We expect to learn how TDP-43 causes toxicity in yeast and in what ways this relates to TDP-43 toxicity in flies, primary cortical neurons and mice. One of our goals is to investigate the range of condensates, oligomers and aggregates formed by TDP-43 and their associated toxicities. Determining which species of TDP-43 is most toxic is an important step towards understanding of toxicity mechanisms. It is also largely unknown what cellular functions are targeted by toxic TDP-43 species and the affiliated mechanisms. We will identify and study cellular targets of toxicity focusing on TDP-43 gain of function toxicity. We will also explore new models of therapeutic approaches by investigating if overexpression of TDP-43 binding proteins can inhibit the formation of toxic TDP-43 species, if titration of important proteins by TDP-43 toxic species contributes to toxicity, and if mutations in TDP-43 can protect WT TDP-43 expressed in the same cell from forming toxic aggregates. Another gap we seek to address is why TDP-43 is associated with different diseases. Importantly, as we showed for yeast prions, TDP-43 and other disease proteins can form distinct aggregate variants (strains), unrelated to mutation, that are associated with distinct characteristics. Thus, different variants of TDP-43 could affect neuronal types differently causing e.g. ALS vs. LATE. TDP-43 variants established in yeast would be important tools to identify disease specific variants and facilitate development of variant specific treatments. We will also investigate the idea that entry into liquid-like granules is an upstream trigger for toxic species formation to learn if liquid-like granules are therapeutic targets. We will quantify the relationship between entry of prion proteins into liquid condensates and stochastic formation of prions in yeast. We will also explore the new area of disease associated metabolite amyloid-like aggregates and the hypothesis that they nucleate prion-like/disease protein misfolding much as we showed cross-seeding between yeast prions. We hope this work will lead to new treatment approaches for protein misfolding diseases.

资助父母补助金的摘要。 某些蛋白质错误折叠，形成与疾病相关的自我播种朊病毒样聚集体。我们专注于一个 这样的蛋白质，TDP-43，因为它是几种神经元聚集体中的主要蛋白质相关的神经元聚集体。 神经变性疾病，包括肌萎缩侧索硬化症（ALS）、额颞叶痴呆和LATE。 LATE是最近描述的一种普遍的TDP-43蛋白病，其导致痴呆，经常被误诊 阿尔茨海默病（AD）。此外，TDP-43存在于与AD和帕金森病相关的聚集体中。 由于TDP-43在酵母中形成聚集体并且是有毒的（抑制生长），因此寻找治疗药物的有效方法是一种有效的方法。 目标是鉴定改变TDP-43毒性的酵母基因。酵母模型与 人类疾病是明确的，因为几个酵母基因，修改人类错误折叠疾病蛋白质的毒性， 包括TDP-43，是新的或已知的人类疾病危险因子的同系物。我们会继续研究 TDP-43聚集体在酵母中的发生和毒性基于我们对酵母自接种朊病毒的专业知识 proteins.我们希望了解TDP-43如何在酵母中引起毒性，以及这与TDP-43的关系 对苍蝇、初级皮层神经元和小鼠的毒性。我们的目标之一是研究冷凝物的范围， TDP-43形成的低聚物和聚集体及其相关毒性。确定哪些物种 TDP-43毒性最强是了解毒性机制的重要一步。它也在很大程度上 目前还不清楚毒性TDP-43物质和相关机制针对哪些细胞功能。我们将 鉴定和研究毒性的细胞靶点，重点是TDP-43功能毒性的获得。我们还将探索 通过研究TDP-43结合蛋白的过表达是否可以 如果TDP-43毒性物质滴定重要蛋白质，则抑制毒性TDP-43物质的形成 如果TDP-43的突变可以保护在同一细胞中表达的WT TDP-43， 形成有毒的聚集体。我们寻求解决的另一个差距是为什么TDP-43与不同的 疾病重要的是，正如我们所展示的酵母朊病毒，TDP-43和其他疾病蛋白可以形成不同的蛋白质。 与突变无关的聚合变体（菌株），与不同特征相关。因此，在本发明中， TDP-43的不同变体可以不同地影响神经元类型，引起例如ALS与LATE。TDP-43变体 在酵母中建立将是重要的工具，以确定疾病特异性变异，并促进发展， 不同的具体治疗。我们还将研究这样的想法，即进入液体状颗粒是一个上游， 触发有毒物质的形成，以了解液体状颗粒是否是治疗目标。我们将量化 朊病毒蛋白进入液体凝聚物和朊病毒在酵母中随机形成之间的关系。 我们还将探索与疾病相关的代谢物淀粉样蛋白样聚集体的新领域， 假设它们使朊病毒样/疾病蛋白质错误折叠成核，就像我们显示的交叉播种一样， 酵母朊病毒我们希望这项工作将为蛋白质错误折叠疾病带来新的治疗方法。