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: 7989598
• 负责人: Davis Martin Seelig
• 金额: $ 10.85万
• 依托单位: COLORADO STATE UNIVERSITY
• 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/7989598
• 关键词: Address; Anatomy; Apoptosis; Applications Grants; Area; B-Lymphocytes; Biological Assay; Blood; Bone Marrow; Bone Marrow Cells; Brain; Cell Death; Cell Lineage; Cells; Cessation of life; Chronic Wasting Disease; Clinical; Collaborations; Communicable Diseases; Confocal Microscopy; Deer; Dendritic Cells; Development; Development Plans; Diagnostic; Disease; Doctor of Philosophy; Documentation; Domestic Animals; Drug or chemical Tissue Distribution; Elements; Employment; Equine mule; Evaluation; Event; Excretory function; Extramural Activities; Flow Cytometry; Genitourinary system; Genomics; Glial Fibrillary Acidic Protein; Gliosis; Goals; Hematogenous; Hematopoietic; Horizontal Disease Transmission; Human; ITGAM gene; Immunofluorescence Immunologic; Immunohistochemistry; In Situ; Infection; Institution; International; Intervention; Intravenous; Investigation; Journals; Kinetics; Knowledge; Label; Laboratories; Lesion; Manuscripts; Marrow; Mediating; Mediator of activation protein; Mentors; Microscopic; Modeling; Molecular Profiling; Monitor; Mus; Myelogenous; Necrosis; Nerve Degeneration; Neurodegenerative Disorders; Neuronal Injury; Neurons; Neuropathogenesis; Oral; Pathogenesis; Pathologist; Pathology; Pathway interactions; Pattern; Peer Review; Performance; Peripheral; Peripheral Nerves; Pilot Projects; Population; PrP; Predisposition; Prion Diseases; Prion Pathway; Prions; Process; Publishing; Relative (related person); Research; Research Personnel; Research Training; Risk; Role; Route; Saliva; Salivary Glands; Sampling; Scanning; Simulate; Site; Study Section; Tail; Text; Therapeutic; Time; Tissues; Tongue; Transgenic Mice; Transgenic Organisms; Transplantation; Tropism; Urine; Western Blotting; Work; abstracting; base; career; career development; caspase-3; cell injury; cervid; design; disease transmission; immunocytochemistry; in vivo; insight; intraperitoneal; macrophage; meetings; monocyte; neuron apoptosis; neuron loss; neuropathology; novel; particle; programs; promoter; protein expression; protein transport; relating to nervous system; research study; response; spatial relationship; tissue tropism; trafficking

DESCRIPTION (provided by applicant): Abstract PI: SEELIG, DAVIS MARTIN Project: 1K01AI082173-01A2 Title: Pathogenesis of Chronic Wasting Disease in Transgenic Mice Accession Number: 3245923 ================== NOTICE: THIS ABSTRACT WAS EXTRACTED FROM APPLICATION AND HAS NOT BEEN PROOFED BY AN SRA.WHEN THERE ARE PROBLEMS WITH THE APPLICATION SCANNING PROCESS, THE EXTRACTED TEXT MAY BE INCORRECT OR INCOMPLETE. ================== 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. A critical limitation in the development of prion disease diagnostic and intervention strategies is the lack of knowledge regarding mechanisms of prion-disease neurodegeneration and in vivo prion trafficking. The work proposed in the application aims to address these limitations by evaluating the accumulation patterns of a marker of prion disease (PrPres) in proximity to specific markers of neuronal pathology which may implicate specific cell populations or pathways capable of being reversed or antagonized. Additionally, our studies will inform us regarding a novel site of PrPres accumulation (the bone marrow) and a novel pathway of prion trafficking (the blood), which will enhance the development of prion disease diagnostics and treatment.

描述(由申请人提供)： 摘要 PI：Seelig，Davis Martin项目：1K01AI082173-01A2标题：转基因小鼠慢性消耗性疾病的发病机制登录号：3245923 = 注意：此摘要摘自应用程序，未经SRA校对。如果应用程序扫描过程有问题，提取的文本可能不正确或不完整。 = 这项建议使用慢性消耗性疾病(CWD)的转基因小鼠模型来研究Pron的序贯发病机制以及骨髓在Pron运输和神经侵袭中的作用。这项研究的目标是：(1)通过对PrP基因表达的转基因小鼠(Tg[CerPrP])的PrP基因序列模式和神经病理学的评估，揭示CWD发病机制的多个方面；(2)通过相互移植研究，探讨骨髓及其细胞后代在PrP转运和发病机制中的作用。我们将利用单标记和双标记免疫组织化学(IHC)、免疫印迹(WB)和免疫荧光共聚焦显微镜相结合的方法来检测错误折叠的Prion蛋白(PrPres)与神经病理标志的堆积。为了确定骨髓在PrP运输和血源性神经侵袭中的作用，我们将使用相互过继的骨髓移植来创建两个嵌合小鼠系，这些嵌合小鼠的PrPc表达将限制在造血系或非造血系。我们将通过流式细胞术对这些小鼠进行表征，并通过IHC和WB记录PrPres的顺序积累。我们预计CWD感染的小鼠将在易感性、临床疾病、显微神经病理学和PrPres分布方面模拟自然感染的宫颈。我们假设/预期CWD感染的小鼠将揭示PrPres病的新的神经病理特征，包括胶质细胞增生标记物的表达增加、神经元丢失和PrPres积聚区域附近的细胞凋亡。在过继骨髓移植实验中，我们假设CWD感染的TG[CerPrP]小鼠的骨髓中含有感染性病毒，这可能与髓系细胞有关。此外，我们假设这些细胞将PrPre运送到大脑，作为细胞更新和运输模式的一部分。这些研究的结果将有助于加深对Pron病相关神经病理机制的理解，有助于为抗Pron病的治疗策略提供依据，并探索一条未被研究的Pron传播途径。利用拟议的实验，我的短期目标将是在2010年夏天完成我的博士学位；然后我的长期目标将是致力于成为一名成功的独立研究人员。为实现这些目标而设计的研究职业发展计划的关键要素包括：(1)建立一个咨询/指导小组，以帮助建立和监测职业业绩里程碑；(2)确定并成功聘用具有传染病和/或神经退行性疾病计划的以研究为重点的部门和机构的调查病理学家；(3)参加已建立的普恩和神经退行性疾病中心的外部研究培训机会；(4)提交同行评审的手稿和外部拨款申请，以及(5)扩大责任并参与实验室、部门和国家/国际会议和期刊俱乐部。Prion病诊断和干预策略发展的一个关键限制是缺乏关于Prion病神经退行性变和体内Prion转运机制的知识。申请中提出的工作旨在通过评估Prion病的标志物(PrPres)在神经元病理的特定标志物附近的积累模式来解决这些限制，这些标志物可能涉及特定的细胞群体或能够逆转或拮抗的途径。此外，我们的研究将使我们了解PrPres积聚的新部位(骨髓)和PrPres运输的新途径(血液)，这将促进PrPre疾病诊断和治疗的发展。