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

描述（由申请人提供）：该项目的目标是增加我们对朊病毒传播的理解，这将有助于我们了解朊病毒病理学并找到这些疾病的治疗方法。朊病毒疾病包括一系列影响人类和许多商业上重要的动物的遗传性、自发性和传播性神经病理。目前，还没有已知的治疗或治愈朊病毒疾病，部分原因是朊病毒病理学的机制和朊病毒繁殖所需的过程还没有很好地理解。我们对朊病毒致病机理的理解将通过识别朊病毒传播所必需的辅助因子而大大增强。大量证据（包括转基因小鼠的研究、细胞培养感染试验、酵母朊病毒繁殖中的类似因素以及体外转化试验）表明，除了朊病毒蛋白的一级序列和表达水平之外，还有其他因素影响朊病毒的繁殖。然而，尽管经过多年的研究，我们对哺乳动物这一过程的了解仍然有限。在某种程度上，缺乏进展可能是由于基于个体的低通量研究来选择候选物。在目标一，我们将利用RNA干扰的力量，以确定潜在的因素，调节朊病毒繁殖的高通量细胞培养试验，并确认我们最有希望的打击敲除小鼠。成功鉴定哺乳动物朊病毒繁殖因子不仅可以提供新的药物靶点，但这可能使我们能够解释为什么尽管朊病毒蛋白表达水平很高，但某些细胞系对朊病毒感染仍然抵抗。在目标二，我们将在一组被几种朊病毒株感染的细胞系上使用RNAi，以鉴定这些潜在的朊病毒繁殖因子中哪些是朊病毒繁殖所必需的，以及这些因子中哪些是朊病毒繁殖所必需的。只对特定的菌株是必需的在目标三中，我们将验证 这些潜在的朊病毒传播因子的子集使用现有的小鼠敲除系。越来越多的证据表明，其他神经退行性疾病是由朊病毒样机制引起的，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.