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: 10725083
• 负责人: SUSAN W LIEBMAN
• 金额: $ 10.61万
• 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/10725083
• 关键词: Address; Affect; Alzheimer' s Disease; Amyloid; Amyotrophic Lateral Sclerosis; Area; Binding Proteins; Cell physiology; Cells; Characteristics; Cytoplasmic Granules; Dementia; Development; Disease; Frontotemporal Dementia; Gene Modified; Growth; Human; Learning; Liquid substance; Methods; Modeling; Mutation; Neurodegenerative Disorders; Neurons; Organism; Parkinson Disease; Physical condensation; PrP; Prions; Proteins; Risk Factors; Therapeutic; Titrations; Toxic effect; Variant; Work; Yeast Model System; Yeasts; cellular targeting; disorder risk; gain of function; human disease; insight; novel therapeutic intervention; 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的普恩样聚集体。我们只关注一件事 这种蛋白质，TDP-43，是几种神经退行性变中与神经元聚集相关的主要蛋白质 疾病，包括肌萎缩侧索硬化症(ALS)、额颞痴呆和晚期。一种流行的TDP- 蛋白沉着症，迟发性痴呆常误诊为阿尔茨海默病(AD)。TDP-43也是 在阿尔茨海默病和帕金森氏症神经元包涵体中发现了其他蛋白质。TDP-43和其他人类错误折叠- ING疾病蛋白在酵母中形成聚集体并抑制生长。这使得酵母可以用来寻找治疗方法- 抽动目标。值得注意的是，酵母基因可以改变人类错误折叠疾病蛋白的毒性，包括 TDP-43，确定了以前未知和已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。我们使用了我们的专业知识，酵母自播种普鲁恩PRO- 利用酵母模型研究人类错误折叠疾病蛋白。现在，我们希望了解TDP-43是如何 在酵母中引起毒性，并在合作者的帮助下，我们的发现以什么方式与TDP-43毒性有关 在高等细胞和生物体中。一项中心任务是确定凝析油、齐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毒性最大将在- 毒性机制的立场。由于有毒的TDP-43以什么细胞功能为靶标在很大程度上是未知的 物种或相关机制，这项工作探索毒性的细胞靶点，重点是TDP-43的获得 功能毒性。新的治疗方法模式将通过调查是否过度表达而开发 TDP-43结合蛋白可以抑制有毒TDP-43物种的形成，如果滴定必不可少或重要 TDP-43有毒物种的细胞蛋白有助于毒性，如果TDP-43的突变可以保护野生动物- TDP-43型在同一细胞中表达，形成有毒聚集体。另一个需要解决的差距是为什么 TDP-43与不同的疾病有关。重要的是，正如我们所展示的酵母蛋白，TDP-43和其他 疾病蛋白可以错误地折叠成不同的自播种聚合体变体/菌株(不是由于其 初级序列)，它们具有明显的特征。因此，TDP-43的不同变体可能会影响神经元 不同的是，导致肌萎缩侧索硬化症和迟发性肌萎缩侧索硬化症。在酵母中建立的TDP-43变异体将成为鉴定TDP-43突变体的重要工具。 确定特定疾病的变种，促进不同的特定治疗方法的发展。我们还将调查 假设进入类液体颗粒是有毒物质形成的上游触发因素，以了解液体- LIKE颗粒是治疗的靶点。我们的方法将是量化Prion进入之间的关系 蛋白质转化为液体冷凝物和酵母菌中随机形成的普恩。最后，我们将研究新的 疾病相关代谢物淀粉样聚集体的面积和它们核化普恩的假说- LIKE/疾病蛋白错误折叠，类似于我们早期演示的酵母普鲁恩之间的交叉播种。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。