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: 8261886
• 负责人: Davis Martin Seelig
• 金额: $ 5.89万
• 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 2012-12-09
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8261886
• 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 distribution; 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）： 抽象的 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：此摘要是从申请中提取的，未经 SRA 验证。当申请扫描过程出现问题时，提取的文本可能不正确或不完整。 ================= 该提案采用慢性消耗性疾病（CWD）的转基因小鼠模型来解决顺序朊病毒发病机制以及骨髓在朊病毒运输和神经侵袭中的作用。拟议研究的目标是：（1）通过评估朊病毒组织趋向性和神经病理学的连续模式来表征转基因、表达鹿 PrP（Tg[CerPrP]）小鼠中 CWD 发病机制的多个方面； (2) 通过相互移植研究来解决骨髓及其细胞后代在朊病毒运输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)、蛋白质印迹 (WB) 和免疫荧光共聚焦显微镜的组合来检测错误折叠朊病毒蛋白 (PrPres) 的积累以及神经病理学标记物。为了确定骨髓在朊病毒运输和血源性神经侵袭中的作用，我们将使用相互过继骨髓移植来创建两个嵌合小鼠谱系，这些小鼠的 PrPc 表达仅限于造血或非造血区室。我们将通过流式细胞术表征这些小鼠，并通过 IHC 和 WB 记录连续的 PrPres 积累。我们预计感染 CWD 的小鼠在易感性、临床疾病、微观神经病理学和 PrPres 分布方面将模仿自然感染的鹿科动物。我们假设/预期感染 CWD 的小鼠将揭示朊病毒病的新的神经病理学特征，包括与 PrPres 积累区域相邻的神经胶质增生标记物表达增加、神经元损失和细胞凋亡。在过继骨髓移植实验中，我们假设 CWD 感染的 Tg[CerPrP] 小鼠的骨髓将含有感染性朊病毒，其与骨髓谱系细胞相关。此外，我们假设这些细胞会将 PrPres 运送到大脑，作为细胞更新和运输模式的一部分。这些研究的结果将增进对朊病毒疾病相关神经病理学机制的理解，有助于为抗朊病毒疾病治疗策略提供信息，并探索尚未得到充分研究的朊病毒传播途径。使用所提议的实验，我的短期目标是在 2010 年夏天完成我的博士学位；我的长期目标将是致力于成为一名成功的独立研究员。旨在实现这些目标的研究职业发展计划的关键要素包括：（1）建立咨询/指导小组，帮助建立和监控职业绩效里程碑； (2) 在具有完善的传染病和/或神经退行性疾病项目的以研究为重点的部门和机构中识别并成功聘用为研究病理学家； (3) 参与已建立的朊病毒和神经退行性疾病中心的校外研究培训机会； (4) 提交同行评审的手稿和校外资助申请，以及 (5) 扩大责任并参与实验室、部门、国家/国际会议和期刊俱乐部。朊病毒疾病诊断和干预策略发展的一个关键限制是缺乏有关朊病毒疾病神经变性和体内朊病毒贩运机制的知识。该申请中提出的工作旨在通过评估朊病毒病标记物（PrPres）在神经元病理学特定标记物附近的积累模式来解决这些局限性，这可能暗示能够逆转或拮抗的特定细胞群或途径。此外，我们的研究将告诉我们关于 PrPres 积累的新位点（骨髓）和朊病毒运输的新途径（血液），这将促进朊病毒疾病诊断和治疗的发展。