Yeast as a gateway to conquering protein misfolding diseases.

抽象的。 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 关注一种这样的蛋白质，TDP-43，因为它是与神经元相关的主要蛋白质 包括肌萎缩侧索硬化症在内的几种神经退行性疾病中的聚集体 (肌萎缩侧索硬化症)，额颞叶痴呆和迟发性。Late是最近被描述的一种流行的TDP-43 导致痴呆症的蛋白质病，通常被误诊为阿尔茨海默病(AD)。在……里面 此外，在与AD和帕金森病相关的聚集体中也发现了TDP-43。 在酵母中形成聚集体并有毒(抑制生长)，这是寻找治疗方法的有效途径 目标一直是识别修改TDP-43毒性的酵母基因。酵母菌的相关性 人类疾病的模型是清楚的，因为几个酵母基因可以改变人类的毒性 错误折叠的疾病蛋白，包括TDP-43，是新的或已知的人类疾病的同源物 风险因素。我们将继续研究酵母中TDP-43聚集体的产生和毒性 基于我们在酵母自种蛋白方面的专业知识。我们希望了解TDP-43是如何 在酵母菌中引起毒性，以及这与果蝇的TDP-43毒性有何关系 神经元和老鼠。我们的目标之一是研究凝析油、齐聚物和 TDP-43形成的聚集体及其相关毒性。确定哪些物种 TDP-43的毒性最大，这是了解毒性机制的重要一步。它是 同样在很大程度上还不清楚有毒的TDP-43物种以什么细胞功能为目标 附属机制。我们将以TDP-43为重点识别和研究细胞毒性靶点 功能毒性的增加。我们还将通过以下方式探索新的治疗方法模式 研究过表达TDP-43结合蛋白是否可以抑制毒性TDP-43的形成 43种，如果TDP-43有毒物种滴定重要蛋白质有助于毒性，以及如果 TDP-43基因突变可保护在同一细胞中表达的WT TDP-43不形成毒性 集合体。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蛋白一样，TDP-43和其他疾病蛋白可以 形成与突变无关的不同聚合体变异(株)，这些变异与不同的 特点。因此，TDP-43的不同变体可能会不同地影响神经元类型，从而导致 例如，肌萎缩侧索硬化症与迟发性脊柱炎。在酵母中建立的TDP-43变异体将是鉴定 疾病特定变种，并促进不同特定治疗方法的开发。我们还将 研究进入类似液体的颗粒是有毒物质上游触发因素的观点 形成以了解液体状颗粒是否为治疗靶点。我们将对这种关系进行量化 普恩蛋白进入液体冷凝物与普恩随机形成之间的关系 酵母。我们还将探索疾病相关代谢物淀粉样蛋白的新领域 聚集体及其核化类病毒/疾病蛋白错误折叠的假说 我们展示了酵母普鲁恩之间的交叉播种。我们希望这项工作将带来新的治疗方法 蛋白质错误折叠疾病的治疗方法。

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.