Project 2

摘要 慢性消耗性疾病(CWD)影响北美和 北欧是一个严重的问题，因为它继续失控地传播。 在野生和圈养的鹿群中。CWD看起来非常不同，有多个不同的 菌株，并可传播给其他动物物种。慢性萎缩性脑病传染给人类的风险 是未知的，这是一个主要的担忧，因为患病动物的数量和它们的地理位置 分布正在迅速增加。CWD在人类之间高效传播的机制 鹿类也是未知的。 这个项目的主要目标是利用一套高度创新的技术来研究细胞， CWD自然发生株的分子和结构特征及其相互作用潜力 物种传播，特别关注某些CWD菌株可能感染的可能性 人类。我们还将尝试阐明CWD普里子的原子分辨结构 低温电子显微镜。最重要的假设是CWD以多个菌株存在于 不同的个体，甚至在同一个个体的不同脑细胞中， 物种传播和人畜共患病的潜力取决于特定的菌株特征。这个 项目分为以下几个具体目标：(1)研究海藻的结构和分子多样性 自然的CWD菌株和CWD蛋白的高分辨率三维结构。(2) 了解激光捕获分离的单个脑细胞中CWD病毒株的多样性 显微切割后用PMCA扩增。(3)CWD的种间评价 传播溢出潜力及其对人畜共患病潜力的影响。(4)鹿人分析 利用含有人类和宫颈神经细胞的嵌合小鼠体内的Pron物种屏障。 包括在这个项目中的研究将解决一些最紧迫的问题 CWD，包括(I)CWD病毒株的变异性，(Ii)不同CWD的人畜共患病潜力 Pron菌株，(Iii)传染性Pron的原子分辨结构和结构基础 (四)CWD病毒株在脑内的细胞分布及其基因表达 后果，(五)慢性萎缩性疾病对其他动物物种的溢出潜力，(六) 在慢性萎缩性脑病病毒向人类传播过程中的中间物种。 在这个项目中产生的结果将是设计防止进一步 传播慢性萎缩性脑病，避免出现新的具有潜在灾难性的疾病 后果。

ABSTRACT Chronic wasting disease (CWD) affecting various species of cervids in North American and Northern Europe represents a serious problem, because it continues to propagate uncontrollably among wild and captive cervids. CWD appears to be very heterogeneous with multiple different strains and can be transmitted to other animal species. The risk of CWD transmission to humans is unknown which is a major concern because the number of sick animals and their geographical distribution is rapidly increasing. The mechanism by which CWD propagates so efficiently among cervids is also unknown. The main goal of this project is to utilize a set of highly innovative techniques to study the cellular, molecular and structural features of naturally occurring CWD strains and their potential for inter- species transmission, particularly focusing on the possibility that certain CWD strains may infect humans. We will also attempt to elucidate the atomic resolution structure of CWD prions using cryo-electron microscopy. The overarching hypothesis is that CWD exists as multiple strains in distinct individuals and even within the same individual in different brain cells and that inter- species transmission and zoonotic potential depend on the specific strain characteristics. The project is divided in the following specific aims: (1) Study the structural and molecular diversity of natural CWD strains and the high resolution three-dimensional structure of CWD prions. (2) Understand CWD prion strain diversity in single brain cells isolated by laser capture microdissection and subsequently amplified by PMCA. (3) Evaluate CWD inter-species transmission spillover potential and its effect on zoonotic potential. (4) Analyze the deer-human prion species barrier in vivo using chimeric mice harboring human and cervid neuronal cells. The studies included in this projects will address some of the most pressing questions regarding CWD, including (i) the CWD prion strain variability, (ii) the zoonotic potential of different CWD prion strains, (iii) the atomic resolution structure of infectious prions and the structural basis of prion strains, (iv) the cellular distribution of CWD prion strains in the brain and its gene expression consequences, (v) the spillover potential of CWD to other animal species, (vi) the potential role of intermediate species in the transmission of CWD prions to humans. The findings generated in this project will be essential to design measures to prevent further propagation of CWD, and to avoid the emergence of new diseases with potentially disastrous consequences.