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.

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 这种蛋白质TDP-43是与几种神经退行性疾病中的神经元聚集体相关的主要蛋白质， 这些疾病包括肌萎缩侧索硬化症（ALS）、额颞叶痴呆和LATE。一个普遍的TDP- 43蛋白质病，LATE导致痴呆常被误诊为阿尔茨海默病（AD）。TDP-43也是 与其他蛋白质一起沿着在AD和帕金森神经元内含物中发现。TDP-43和其他人类错误折叠- 使疾病蛋白质形成聚集体并抑制酵母的生长。这使得酵母可以用来寻找治疗方法， 目标。值得注意的是，酵母基因修改人类错误折叠疾病蛋白质的毒性，包括 TDP-43，确定了以前未知的和确定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。我们利用我们的专业知识与酵母自我播种朊病毒亲， 用酵母模型研究人类错误折叠疾病蛋白。我们希望了解TDP-43 在合作者的帮助下，我们的研究结果与TDP-43毒性的关系如何？ 在高等细胞和有机体中。一个中心任务是确定冷凝物，低聚物和聚集体的范围 由TDP-43形成，以及它们的毒性。确定哪种TDP-43物质毒性最大将在以下条件下进行- 毒性机制。由于很大程度上尚不清楚有毒的TDP-43针对的细胞功能是什么 物种或附属机制，这项工作探讨了毒性的细胞靶点，重点是TDP-43获得 功能毒性新的治疗方法模型将通过研究是否过度表达 TDP-43结合蛋白可抑制毒性TDP-43物质的形成，如果滴定必需或重要 TDP-43毒性物种的细胞蛋白有助于毒性，如果TDP-43突变可以保护野生型， TDP-43型在同一细胞中表达，形成毒性聚集体。另一个需要解决的差距是， TDP-43与不同的疾病有关。重要的是，正如我们在酵母朊病毒中所展示的，TDP-43和其他 疾病蛋白质可以错误折叠成不同的自接种聚集体变体/菌株（不是由于它们的结构改变， 主要序列），具有不同的特征。因此，TDP-43的不同变体可以影响神经元 不同地，导致例如ALS与LATE。在酵母中建立的TDP-43变异体将是识别 确认疾病特异性变体，并促进变体特异性治疗的开发。我们亦会研究 假设进入液体状颗粒是有毒物质形成的上游触发因素，以了解液体- 类似颗粒是治疗靶点。我们的方法将是量化朊病毒进入 蛋白质转化为液体凝聚物和朊病毒在酵母中的随机形成。最后，我们将研究新的 与疾病相关的代谢物淀粉样聚集体的区域以及它们使朊病毒成核的假设， 类似/疾病蛋白质错误折叠，类似于我们早期展示的酵母朊病毒之间的交叉接种。这 这项工作有望为蛋白质错误折叠疾病带来新的治疗方法。