Pathogenesis of Chronic Wasting Disease in Transgenic Mice

转基因小鼠慢性消耗性疾病的发病机制

• 批准号: 8614388
• 负责人: Davis Martin Seelig
• 金额: $ 5.2万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8614388
• 关键词: Pathogenesis; Chronic; Wasting; Disease; Transgenic

Project Summary/Abstract: This proposal employs a transgenic murine model of chronic wasting disease (CWD) to addresses sequential prion pathogenesis and the role of bone marrow in prion trafficking and neuroinvasion. The goals of the proposed studies are: (1) to characterize the multiple aspects of the pathogenesis of CWD in transgenic, cervid PrP-expressing (Tg[CerPrP]) mice by evaluation of sequential patterns of prion tissue tropism and neuropathology; and (2) to address the role of the bone marrow and its cellular progeny in the prion trafficking and pathogenesis through reciprocal transplantation studies. We will utilize a combination of single- and dual- labeling immunohistochemistry (IHC), western blotting (WB), and immunofluorescent confocal microscopy to detect accumulations of misfolded prion protein (PrPres) in conjunction with markers of neuropathology. To define the role of the bone marrow in prion trafficking and hematogenous neuroinvasion, we will use reciprocal adoptive bone marrow transfer to create two lineages of chimeric mice, which will have PrPc expression restricted to hematopoietic or non-hematopoietic compartments. We will characterize these mice through flow cytometry, and document sequential PrPres accumulation via IHC and WB. We expect that the CWD-infected mice will mimic naturally-infected cervids with respect to susceptibility, clinical disease, microscopic neuropathology, and PrPres distribution. We hypothesize/anticipate that CWD-infected mice will reveal novel, neuropathologic features of prion disease, including increased expression of markers of gliosis, neuronal loss and apoptosis adjacent to areas of PrPres accumulation. In the adoptive bone marrow transfer experiments, we hypothesize that marrow of CWD-infected Tg[CerPrP] mice will contain infectious prions, which will be associated with myeloid lineage cells. Additionally, we hypothesize that these cells will ferry PrPres to the brain as part of cell renewal and trafficking patterns. The results of these studies will enhance understanding of the mechanisms of prion disease-associated neuropathology, help inform anti-prion disease therapeutic strategies, and explore an under-investigated pathway of prion dissemination. Using the proposed experiments, my short- term objective will be to complete my PhD in the summer of 2010; my long term goal will then be to devote myself to a becoming a successful independent researcher. The key elements of the research career development plan designed to accomplish these goals include: (1) the establishment of an advisory/mentoring panel to help establish and monitor career performance milestones; (2) the identification and successful employment as an investigative pathologist in a research-focused department and institution with a well- established infectious disease and/or neurodegenerative program; (3) participation in extramural research training opportunities at established centers of prion and neurodegenerative disease; (4) submission of peer- reviewed manuscripts and extramural grant applications, and (5) extended responsibility and participation in laboratory, departmental, and national/international meetings and journal clubs.

项目概要/摘要： 该建议采用慢性消耗性疾病（CWD）的转基因小鼠模型来解决连续的 朊病毒的发病机制以及骨髓在朊病毒运输和神经侵袭中的作用。的目标 建议的研究是：（1）在转基因、鹿、猪、 PrP表达（Tg[CerPrP]）小鼠，通过评价朊病毒组织嗜性的顺序模式， 神经病理学;和（2）解决骨髓及其细胞后代在朊病毒运输中的作用 和发病机制的研究。我们将使用单和双- 标记免疫组织化学（IHC）、蛋白质印迹（WB）和免疫荧光共聚焦显微镜， 结合神经病理学标志物检测错误折叠朊病毒蛋白（PrPres）的积累。到 为了明确骨髓在朊病毒运输和血行性神经侵袭中的作用，我们将采用相互作用的方法， 过继骨髓转移以产生两个谱系的嵌合小鼠，其将具有PrPc表达 仅限于造血或非造血区室。我们将通过流动来描述这些小鼠的特征， 流式细胞术，并通过IHC和WB记录连续的PrPres积累。我们认为感染了化学武器致死病的人 小鼠将在易感性、临床疾病、显微镜检查 神经病理学和PrPres分布。我们假设/预期感染CWD的小鼠将揭示新的， 朊病毒病的神经病理学特征，包括神经胶质增生标志物表达增加，神经元丢失 和与PrPres聚集区域相邻的凋亡。在骨髓移植实验中，我们 假设感染CWD Tg[CerPrP]小鼠的骨髓将含有感染性朊病毒， 与骨髓谱系细胞相关。此外，我们假设这些细胞将PrPres运送到大脑， 作为细胞更新和贩运模式的一部分。这些研究的结果将增进对 朊病毒病相关神经病理学的机制，有助于告知抗朊病毒病治疗策略， 探索朊病毒传播的一条未被充分研究的途径。通过这些实验，我的短- 我的长期目标是在2010年夏天完成我的博士学位;我的长期目标是致力于 成为一名成功的独立研究员。研究生涯的关键要素 旨在实现这些目标的发展计划包括：（1）建立咨询/指导 小组，以帮助建立和监测职业绩效里程碑;（2）确定和成功的 在一个以研究为重点的部门和机构担任调查病理学家， 已建立的传染病和/或神经退行性疾病项目;（3）参与校外研究 在已建立的朊病毒和神经退行性疾病中心的培训机会;（4）提交同行- 审查手稿和校外赠款申请，（5）扩大责任和参与， 实验室，部门和国家/国际会议和期刊俱乐部。