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

描述(申请人提供)：传播性海绵状脑病(TSE)是一组影响人类和动物的致命神经退行性疾病。虽然罕见，但最近牛海绵状脑病(BSE)和宫颈慢性衰弱病(CWD)的爆发以及疾病从牛到人的传播引起了人们的极大关注。由于缺乏有效、敏感和早期的诊断，以及围绕这种传染病前所未有的性质、其传播机制和控制普恩传播的物种屏障的许多不确定性，这一问题变得更加严重。该计划项目的主要目标是了解疯牛病和慢性萎缩性脑病病毒的传播机制和发病机制，估计这些病原体将疾病传播给其他动物特别是人类的风险，评估病毒在动物中传播的机制和途径，并开发新的动物死亡前检测策略。一个由在该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通力合作，以实现这些目标。项目1(Juergen Richt，PL)建议详细研究突变和淘汰奶牛，并评估疯牛病遗传形式在疾病传播、易感性和物种障碍中的作用。对于这些研究，我们计划培育和表征最近在美国一例自然疯牛病病例中发现的表达PrP突变的敲打转基因牛。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 Tell，PL)建议建立和使用转基因小鼠模型，以评估各种物种屏障的强度和Pron菌株的影响。我们将研究不同模型感染CWD的敏感性，以及鹿模型可能感染其他物种的普恩病毒的可能性。为了达到这一目标，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(Cldio Soto，PL)建议研究CWD和BSE Prion的体外复制，评估感染蛋白的组织分布，揭示传播途径，并开发诊断方法。为此，我们将使用索托博士实验室最近开发的PMCA技术来模拟Pron复制过程。行政核心(核心A)(克劳迪奥·索托主任)将促进和整合项目和核心，并处理方案顺利运作所需的所有行政方面。组织核心(核心B)(皮尔路易吉·甘贝蒂主任)将处理、分析和存储产生的组织样本，并为成员提供进入生物安全设施的途径，以操纵疯牛病传染病材料。本计划所产生的结果无疑将有助于理解这两种令人担忧的人兽共患病的发病机制、传播途径和早期发现。

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.