Pathogenesis, Transmission and Detection of Zoonotic Prion Diseases (P01)

描述（由申请方提供）：传染性海绵状脑病（TSE）是一组影响人类和动物的致死性神经退行性疾病。虽然罕见，但最近在牛中爆发的牛海绵状脑病（BSE）和在鹿中爆发的慢性消耗病（CWD）以及该疾病从牛到人的传播引起了极大的关注。由于缺乏有效、敏感和早期的诊断方法，以及围绕着这种传染性病原体的前所未有的性质、其传播机制和控制朊病毒传播的物种屏障的许多不确定性，这一问题更加严重。本计划项目的主要目标是了解疯牛病和慢性消耗病朊病毒的传播和发病机制，估计这些传染性病原体传播疾病给其他动物，特别是人类的风险，评估朊病毒在动物中传播的机制和途径，并开发新的策略，用于受感染动物的死前检测。一个由在这一领域具有广泛公认的专业知识和贡献记录的有成就的科学家组成的团队将协同工作，以实现这些目标。项目1（Juergen Richt，PL）建议详细研究突变和敲除奶牛，并评估BSE遗传形式在疾病传播、易感性和物种屏障中的作用。对于这些研究，我们计划在转基因牛中产生和表征敲入，该转基因牛表达最近在美国的一个自然BSE病例中鉴定的PrP突变。我们还将使用表达野生型和突变型牛PrP的转基因小鼠模型，并使用PMCA进行PrP复制的体外研究。项目2（Glenn Telling，PL）建议生成和使用转基因小鼠模型来评估各种物种屏障的强度和朊病毒株的影响。我们将研究各种模型被慢性消耗病感染的易感性，以及鹿模型可能被其他物种的朊病毒感染的可能性。为了实现这一目标，将在转基因小鼠模型中进行实验，并使用体外转化研究。项目3（Claudio Soto，PL）建议研究体外CWD和BSE朊病毒的复制，评估感染性蛋白的组织分布，揭示传播途径并开发诊断检测方法。为此，我们将使用最近在Soto博士实验室开发的PMCA技术来模拟朊病毒复制过程。行政核心（核心A）（Claudio Soto，主任）将促进和整合项目和核心，并负责计划顺利运行所需的所有行政方面。组织核心（核心B）（Pierluigi Gambetti，负责人）将处理、分析和储存生成的组织样本，并为成员提供进入生物安全机构的通道，以处理BSE感染性材料。该项目的发现无疑将有助于了解这两种令人担忧的人畜共患朊病毒疾病的发病机制、传播途径和早期检测。

DESCRIPTION (provided by applicant): Transmissible spongiform encephalopathies (TSEs) are a group of fatal neurodegenerative disorders affecting humans and animals. Although rare, the recent outbreak of Bovine Spongiform Encephalopathy(BSE) in cattle and Chronic Wasting disease (CWD) in cervids and the transmission of the disease from cattle to humans have caused great concern. This problem is aggravated by the lack of an efficient, sensitive and early diagnosis as well as many uncertainties surrounding the unprecedented nature of the infectious agent, its mechanism of propagation and the species-barrier that controls prion transmission. The major goals of this Program Project are to understand the mechanism of transmission and pathogenesis of BSE and CWD prions, to estimate the risk of these infectious agents to propagate disease to other animals and especially to humans, to assess the mechanisms and routes of prion dissemination among animals and to develop novel strategies for ante-mortem detection of infected animals. A team of accomplished scientists with widely recognized expertise and track record of contributions in this field will work collaboratively to reach these goals. Project 1 (Juergen Richt, PL) proposes to study in detail mutant and knock out cows and assess the role of genetic forms of BSE in disease transmission, susceptibility and species barrier. For these studies, we plan to generate and characterize knock in transgenic cattle expressing a PrP mutation recently identified in a natural BSE case in USA. We will also use transgenic mice models expressing wild type and mutant bovine PrP and in vitro studies of PrP replication using the PMCA. Project 2 (Glenn Telling, PL) proposes to generate and use transgenic mice models to assess the strength of various species barriers and the influence of prion strains. We will study the susceptibility of various models to be infected by CWD as well as the possibility that deer models may be infected by prions from other species. To address this aim, experiments will be done in transgenic mice models and using in vitro conversion studies. Project 3 (Claudio Soto, PL) proposes to study the replication of CWD and BSE prions in vitro, evaluate tissue distributions of infectious protein, enlighten the routes of transmission and develop a diagnostic assay. For this purpose, we will use the PMCA technology, recently developed in Dr Soto's lab to mimic the prion replication process. The Administration Core (core A) (Claudio Soto, Director) will facilitate and integrate Projects and Cores and take care of all administrative aspects needed for the smooth operation of the Program. The Tissue Core (core B) (Pierluigi Gambetti, Director) will process, analyze and store the tissue samples generated and provide access to the members to a biosafety facility to manipulate BSE infectious material. The findings generated in this Program will have undoubtedly contribute to understand the pathogenesis, route of propagation and early detection of these two worrisome zoonotic prion diseases.