Identifying the Role of PrPSc Distribution & Other Proteins in Prion Pathogenesis

描述(申请人提供)：这个项目的目标是增加我们对Pron传播的了解，这将有助于我们了解Pron的病理学，并找到这些疾病的治疗方法。Pron疾病包括一系列遗传的、自发的和传播的神经病理，影响人类和一些具有重要商业意义的动物。目前，还没有已知的治疗或治愈Pron疾病的方法，部分原因是Pron病理基础的机制和Pron传播所需的过程还不是很清楚。通过确定PrP繁殖所必需的辅助因子，我们对Pron发病机制的了解将会大大提高。大量证据(包括在转基因小鼠中的工作、细胞培养感染分析、酵母蛋白繁殖中的类似因素以及体外转化试验)表明，除了Prion蛋白的初级序列和表达水平之外，还有其他因素影响Prion的繁殖。然而，尽管进行了多年的研究，我们对哺乳动物体内这一过程的了解仍然有限。在一定程度上，缺乏进展可能是因为候选人的选择是基于个人的低吞吐量研究。在第一个目标中，我们将利用RNA干扰的力量来识别在高通量细胞培养分析中调节普恩传播的潜在因素，并确认我们在基因敲除小鼠中最有希望的点击。成功识别哺乳动物的Prion繁殖因子不仅将提供新的药物靶点，而且可能使我们能够解释为什么一些细胞系在Prion蛋白高水平表达的情况下仍然抵抗Prion感染。在目标2中，我们将在一组感染了几种Pron菌株的细胞系上使用RNAi来识别这些潜在的Prion繁殖因子中的哪些总是必需的，以及这些因子中的哪些只对特定菌株是必需的。在目标三中，我们将验证一个 利用现有的小鼠基因敲除系获得这些潜在的普恩病毒传播因子的子集。越来越多的证据表明，其他神经退行性疾病是由类病毒机制引起的，这增加了这些结果可能对人类健康产生广泛影响的可能性。

DESCRIPTION (provided by applicant): The goal of this project is to increase our understanding of prion propagation, which will help us to understand prion pathology and find treatments for these diseases. Prion diseases encompass a range of genetic, spontaneous, and transmitted neuropathologies that affect humans and a number of commercially important animals. At present, there is no known treatment or cure for prion diseases, in part because the mechanisms underlying prion pathology and the processes required for prion propagation are not well understood. Our understanding of prion pathogenesis would be greatly enhanced by identifying the auxiliary factors that are necessary for prion propagation. Numerous lines of evidence (including work in transgenic mice, cell culture infection assays, analogous factors in yeast prion propagation, and in vitro conversion assays) suggest that additional factors besides the primary sequence and expression level of the prion protein influence prion propagation. However, despite years of study, our understanding of this process in mammals is still limited. In part, this lack of progress may be due to the selection of candidates based on individual, low throughput studies. In aim one, we will harness the power of RNA interference to identify potential factors that modulate prion propagation in high throughput cell culture assays, and confirm our most promising hits in knockout mice. Successfully identifying mammalian prion propagation factors would not only provide new drug targets, but may allow us to explain why some cell lines are recalcitrant to prion infection despite high levels of prion protein expression In aim two we will use RNAi on a panel of cell lines infected with several prion strains to identif which of these potential prion propagation factors are always necessary for prion propagation and which of these factors are only necessary for specific strains. In aim three we will validate a subset of these potential prion propagation factors using existing mouse knockout lines. Increasing evidence that other neurodegenerative diseases are caused by a prion-like mechanism raises the possibility that these results could have a broad impact on human health.