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: 10571373
• 负责人: SUSAN W LIEBMAN
• 金额: $ 25.39万
• 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/10571373
• 关键词: Address; Affect; Alzheimer' s Disease; Amyloid; Amyotrophic Lateral Sclerosis; Area; Binding Proteins; Cell physiology; Cells; Characteristics; Cytoplasmic Granules; Dementia; Development; Disease; Gene-Modified; Growth; Human; Lead; Learning; Liquid substance; Methods; Modeling; Mutation; Neurodegenerative Disorders; Neurons; Organism; Parkinson Disease; PrP; Prions; Proteins; Risk Factors; Therapeutic; Titrations; Toxic effect; Variant; Work; Yeast Model System; Yeasts; cellular targeting; disorder risk; frontotemporal lobar dementia-amyotrophic lateral sclerosis; gain of function; human disease; insight; overexpression; prion seeds; prion-like; protein TDP-43; protein aggregation; protein misfolding; therapeutic target; tool; yeast prion

ABSTRACT Certain proteins misfold to form self-seeding prion-like aggregates associated with disease. We focus on one such protein, TDP-43, the major protein associated with neuronal aggregates in several neurodegenerative diseases, including amyotrophic lateral sclerosis (ALS), frontotemporal dementia and LATE. A prevalent TDP- 43 proteinopathy, LATE causes dementia often misdiagnosed as Alzheimer’s disease (AD). TDP-43 is also found along with other proteins in AD and Parkinson’s neuronal inclusions. TDP-43 and other human misfold- ing disease proteins form aggregates and inhibit growth in yeast. This allows yeast to be used to find therapeu- tic targets. Remarkably, yeast genes that modify the toxicity of human misfolding disease proteins, including TDP-43, identified both previously unknown and established human disease risk factors, demonstrating the relevancy of the yeast model to human disease. We have used our expertise with yeast self-seeding prion pro- teins to study human misfolding disease proteins with the yeast model. Now, we expect to learn how TDP-43 causes toxicity in yeast and, with the help of collaborators, in what ways our findings relate to TDP-43 toxicity in higher cells and organisms. A central task is to identify the range of condensates, oligomers and aggregates formed by TDP-43, and their toxicities. Determining which TDP-43 species is most toxic will advance under- standing of toxicity mechanisms. As it is largely unknown what cellular functions are targeted by toxic TDP-43 species or the affiliated mechanisms, this work explores cellular targets of toxicity focusing on TDP-43 gain of function toxicity. New models of therapeutic approaches will be developed by investigating if overexpression of TDP-43 binding proteins can inhibit the formation of toxic TDP-43 species, if titration of essential or important cellular proteins by TDP-43 toxic species contributes to toxicity, and if mutations in TDP-43 can protect wild- type TDP-43 expressed in the same cell from forming toxic aggregates. Another gap to be addressed is why TDP-43 is associated with different diseases. Importantly, as we showed for yeast prions, TDP-43 and other disease proteins can misfold into different self-seeding aggregate variants/strains (not due to alterations in their primary sequence), that have distinct characteristics. Thus, different variants of TDP-43 could affect neurons differently, causing e.g. ALS vs. LATE. TDP-43 variants established in yeast would be important tools to iden- tify disease specific variants and facilitate development of variant specific treatments. We will also investigate the premise that entry into liquid-like granules is an upstream trigger of toxic species formation to learn if liquid- like granules are therapeutic targets. Our approach will be to quantify the relationship between entry of prion proteins into liquid condensates and stochastic formation of prions in yeast. Finally, we will examine the new area of disease associated metabolite amyloid-like aggregates and the hypothesis that they nucleate prion- like/disease protein misfolding, similar to our early demonstration of cross-seeding between yeast prions. This work is expected to lead to new treatment approaches for protein misfolding diseases.

抽象的 某些蛋白质错误折叠以形成与疾病相关的自种群的prion样骨料。我们专注于一个 这种蛋白质，TDP-43，这是几种神经退行性中与神经元聚集体相关的主要蛋白 疾病，包括肌萎缩性侧索硬化症（ALS），额颞痴呆和晚期。普遍的TDP- 43蛋白质病，晚期导致痴呆症通常被误诊为阿尔茨海默氏病（AD）。 TDP-43也是 与AD和Parkinson的神经元内包含中的其他蛋白质一起发现。 TDP-43和其他人类错误折叠 ING疾病蛋白形成聚集体并抑制酵母的生长。这使酵母可用于找到治疗 - 抽动目标。值得注意的是，改变了人类错误折叠疾病蛋白的毒性的酵母基因，包括 TDP-43确定了以前未知和已建立的人类疾病风险因素，证明了 酵母模型与人类疾病的相关性。我们已经将我们的专业知识用于酵母自种群 用酵母模型研究人类错误折叠疾病蛋白的Teins。现在，我们希望了解TDP-43如何 在酵母中引起毒性，并在合作者的帮助下，我们的发现与TDP-43毒性有关 在更高的细胞和组织中。一个核心任务是确定冷凝水，低聚物和聚集体的范围 由TDP-43及其毒性形成。确定哪种TDP-43种最具毒性的物种将推动不足 毒性机制的站立。因为有毒TDP-43的靶向哪些细胞功能在很大程度上尚不清楚 物种或会员机制，这项工作探讨了毒性的细胞靶标，重点是TDP-43 功能毒性。理论方法的新模型将通过研究是否过表达来开发 TDP-43结合蛋白可以抑制有毒TDP-43种的形成，如果滴定为必要或重要 TDP-43的细胞蛋白有毒物种会导致毒性，如果TDP-43中的突变可以保护野生 TDP-43型在形成有毒骨料的同一细胞中表达。要解决的另一个差距是为什么 TDP-43与不同的疾病有关。重要的是，正如我们为酵母菌pr，TDP-43和其他 疾病蛋白可能会误解为不同的自种骨料变体/菌株（不是由于其改变的改变 主要序列），具有不同特征。那，TDP-43的不同变体可能会影响神经元 不同的是，导致例如ALS vs.晚。在酵母中建立的TDP-43变体将是重要的工具 介绍特定疾病的变异并促进变异特定治疗的发展。我们还将调查 进入液状颗粒的前提是有毒物种形成的上游触发器，以了解液体是否存在 像颗粒一样是治疗靶标。我们的方法是量化Prime进入之间的关系 蛋白质成液体冷凝水和酵母中prion的随机形成。最后，我们将研究新的 疾病区域相关的代谢产物淀粉样蛋白样骨料，并假设它们是核质量的 类似于疾病蛋白的错误折叠，类似于我们早期证明酵母菌王室之间交叉种子的证明。这 预计工作将导致蛋白质错误折叠疾病的新治疗方法。