Redefining the zoonotic potential of chronic wasting disease

重新定义慢性消耗性疾病的人畜共患潜力

基本信息

批准号：
10610969
负责人：
Sabine Gilch
金额：
$ 27万
依托单位：
UNIVERSITY OF CALGARY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-06-01 至 2026-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10610969
关键词：
Address American Animals Asia Astrocytosis Biochemical Biological Bovine Spongiform Encephalopathy Brain Cattle Cell Culture Techniques Characteristics Chronic Wasting Disease Clinical Consumption Data Deer Deposition Disease Disease Outbreaks Dissociation Eating Endopeptidase K Europe Euthanasia Exhibits Exposure to Farm Feces Goals Health Human In Vitro Incubated Infection Link Macaca Macaca fascicularis Microtus Modeling Molecular Mus Neurodegenerative Disorders North America Oral Pathologic Pathway interactions Peptide Hydrolases Persons Phenotype PrP PrPSc Proteins Prion Diseases Prions Process Property Protein Fragment Protein Isoforms Public Health Reindeer Research Resistance Risk Rodent Route Saliva Testing Time Tissues Toxic effect Transgenic Mice Urine Zoonoses brain cell cervid disease transmission driving force epidemiologic data experimental study human model humanized mouse in vivo innovation insight mouse model nonhuman primate overexpression prion seeds reconstitution response single-cell RNA sequencing transmission process

项目摘要

The rapid expansion of chronic wasting disease (CWD), a prion disease of free-ranging and farmed deer, elk and moose, is a major and ongoing threat in North America. Approximately 1 in 36 Americans hunt deer and elk and eat venison, and it is estimated that 7,000 – 15,000 CWD-infected cervids are consumed annually. This fuels growing concerns about the human health risks imposed by CWD. There are no documented cases of CWD transmission to humans, even though with the long incubation periods of all prion diseases and the unknown presentation of CWD in humans definite conclusions are not possible. The zoonotic potential of prion diseases has been exemplified by bovine spongiform encephalopathy (BSE, mad cow disease) which resulted in a new form of human prion disease (vCJD). BSE was transmissible to Cynomolgus macaques and transgenic mice expressing the human prion protein. Initial results of CWD transmission studies to the same non-human primate and mouse models of human prion disease were not successful, corroborating the conclusion that the zoonotic potential of CWD is low, if not absent. Our groups were part of a consortium that inoculated Cynomolgus macaques via different routes with CWD. Some animals exhibited subtle clinical signs reminiscent of prion disease, and upon euthanasia, weak signs of vacuolation, PrPSc deposition and astrocytosis in the brain were found, while no proteinase K (PK) resistant prion protein (PrP) was detectable. We have now demonstrated for the first time that CWD from macaques can transmit clinical prion disease to transgenic mouse models of CWD and human prion disease, albeit in the absence of detectable PK-resistant PrP. Bona fide PrPSc was only detected upon 3rd passage from mouse to bank vole models. Altogether, this is the first evidence that CWD very likely has zoonotic potential. The goal of the current proposal is to redefine the zoonotic potential of CWD by characterizing the biological properties of CWD prions emerging upon experimental transmission into macaques, for obtaining important information on how CWD could manifest in humans. In Aim 1, we will study whether CWD from macaque (CWDmac) in bank voles represents a new prion strain, by comparing biochemical and biological properties to an array of known prion strains from different species. Aim 2 addresses the question whether CWDmac represents an intermediate prion strain, adaptable to cervids or humans upon passage, and possessing an expanded host range. We will address this by in vivo passage in cervidized or humanized mouse models. In vitro, we will utilize serial PMCA and a newly generated PrP0/0 cell culture model for infection, upon reconstitution with PrP from different species. In Aim 3, we will shed light on the observed dissociation between infectivity and the presence of bona fide PrPSc. We propose to identify atypical PrP fragments associated with CWDmac, and we will elucidate brain cell responses to CWDmac exposure by innovative single cell RNA sequencing. In summary, our studies will uncover the possible manifestation of CWD in humans, which is of critical importance for drawing definite conclusions about the zoonotic potential of CWD.

慢性浪费疾病（CWD）的快速扩张，这是一种自由放养和养殖鹿的prion病穆斯（Moose）是北美的主要且持续的威胁。大约有36位美国人狩猎鹿和麋鹿并吃鹿肉，据估计，每年消耗7,000至15,000个CWD感染的子宫颈。这燃料日益担心CWD施加的人类健康风险。没有记录的案例 CWD向人类传播，即使所有prion疾病的长期孵化期和在人类中，CWD的未知介绍是不可能的。 Prime的人畜共患潜力牛赞助脑病（BSE，疯牛病）已经举例说明了疾病以新形式的人类prion病（VCJD）。 BSE可传递给cynomolgus猕猴和转基因表达人prion蛋白的小鼠。 CWD传播研究对同一非人类的初步结果人类儿童疾病的灵长类动物和小鼠模型没有成功，证实了这样的结论： CWD的人畜共患潜力很低，即使不存在。我们的小组是接种cynomolgus的财团的一部分通过CWD的不同路线猕猴。一些动物暴露了微妙的临床迹象疾病和安乐死时，大脑中真空吸尘，PRPSC沉积和星形细胞增多症的迹象很弱。发现，虽然未检测到未检测到蛋白酶K（PK）抗性蛋白（PRP）。我们现在已经证明了猕猴的CWD首次将临床prion病传输到CWD的转基因小鼠模型和人类病毒疾病，尽管没有可检测到的PK抗PK的PRP。真正的prpsc只是在从鼠标到银行Vole模型的第三段中检测到。总之，这是CWD非常的证据可能具有人畜共患潜力。当前建议的目的是重新定义CWD的人畜共患潜力通过表征在实验传播上出现的CWD王室的生物学特性进入猕猴，以获取有关CWD如何在人类中表现出来的重要信息。在AIM 1中，我们将通过比较，将研究来自猕猴的CWD（CWDMAC）是否代表一种新的prion菌株来自不同物种的一系列已知prion菌株的生化和生物学特性。 AIM 2地址 CWDMAC是否代表中间pr菌株，适应于子宫颈或人类的问题通道，并具有扩展的主机范围。我们将通过子体内段落中的体内通过或人源化的小鼠模型。在体外，我们将利用序列PMCA和新生成的PRP0/0细胞培养模型为了感染，与来自不同物种的PRP重组后。在AIM 3中，我们将阐明观察到的感染与真正的PRPSC的存在之间的解离。我们建议确定非典型PRP 与CWDMAC相关的片段，我们将通过创新来阐明脑细胞对CWDMAC暴露的反应单细胞RNA测序。总而言之，我们的研究将发现人类中CWD的可能表现，这对于得出有关CWD的人畜共患潜力的明确结论至关重要。