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。

基本信息

批准号：
10810084
负责人：
SUSAN W LIEBMAN
金额：
$ 1.15万
依托单位：
UNIVERSITY OF NEVADA RENO
依托单位国家：
美国
项目类别：
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

项目摘要

Abstract. 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.

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。我们专注于这样一种蛋白质TDP-43，因为它是主要蛋白相关的神经元几种神经退行性疾病（包括肌萎缩性侧硬化症）的聚集体（ALS），额颞痴呆和晚期。迟到是最近描述的普遍的TDP-43 引起痴呆症通常被诊断为阿尔茨海默氏病（AD）的蛋白质病。此外，在与AD和帕金森氏症相关的聚集体中发现了TDP-43。自TDP-43以来形成骨料的形式，有毒（抑制生长）酵母，这是一种有力的治疗方法靶标是鉴定改变TDP-43毒性的酵母基因。酵母的相关性人类疾病的模型很明显，因为几种改变人类毒性的酵母基因包括TDP-43在内的错误折叠疾病蛋白是新的或已知人类疾病的同源物风险因素。我们将继续研究酵母中TDP-43骨料的起源和毒性以酵母自种蛋白质蛋白质为基础我们的专业知识。我们希望了解TDP-43 引起酵母的毒性，并以何种方式与TDP-43毒性有关神经元和小鼠。我们的目标之一是研究冷凝水，低聚物和 TDP-43及其相关毒性形成的聚集体。确定哪种物种的 TDP-43是最具毒性的是了解毒性机制的重要一步。这是同样，有毒TDP-43种针对哪些细胞功能，也未知哪些细胞功能会员机制。我们将识别和研究关注TDP-43的毒性细胞靶标功能毒性的增益。我们还将通过研究TDP-43结合蛋白的过表达是否会抑制有毒TDP-的形成 43种，如果TDP-43对重要蛋白的滴定有毒物种有助于毒性，如果 TDP-43中的突变可以保护在同一细胞中表达的WT TDP-43免于形成有毒聚合。我们寻求解决的另一个差距是为什么TDP-43与不同疾病。重要的是，正如我们对酵母菌pr，TDP-43和其他疾病蛋白的展示形成与突变无关的不同聚集变体（菌株），与突变无关特征。这是TDP-43的不同变体可能会影响神经元类型，从而导致不同例如ALS vs.晚。在酵母中建立的TDP-43变体将是识别的重要工具疾病特定的变异并促进变体特定治疗的发展。我们也会调查进入液状颗粒是有毒物种的上游触发的想法形成以了解液态颗粒是否是治疗靶标。我们将量化关系在将prion蛋白进入液态冷凝水与王室的随机形成之间酵母。我们还将探索与类似淀粉样蛋白的疾病新领域聚集体和它们的核素蛋白蛋白质核蛋白的错误折叠率很大我们在酵母菌王室之间表现出交叉种子。我们希望这项工作将导致新的治疗蛋白质错误折叠疾病的方法。