Investigating the RhoA/ROCK pathway for the treatment of Alzheimer's disease

研究治疗阿尔茨海默病的 RhoA/ROCK 通路

• 批准号: 8898696
• 负责人: Jeremy H. Herskowitz
• 金额: $ 24.15万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2017-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8898696
• 关键词: Address; Affect; Alzheimer disease prevention; Alzheimer' s Disease; American; Amino Acid Substitution; Amyloid; Autopsy; Award; Basic Science; Binding; Biological; Biological Models; Brain; Cells; Cellular biology; Clinical; Cognition; Coupled; Data; Dementia; Dendritic Spines; Dependovirus; Deposition; Development; Disease; Disease Progression; Drug Targeting; Education; Ethics; Event; Experimental Models; Faculty; Family member; Fostering; Functional disorder; Future; Generations; Goals; Hippocampus (Brain); Human; Impaired cognition; Injection of therapeutic agent; International; Intervention; Knowledge; Laboratories; Lead; Leadership; Learning; Marines; Measures; Mediating; Mentors; Methodology; Microscopy; Modeling; Molecular; Morphology; Mus; Nerve Degeneration; Neurites; Neurodegenerative Disorders; Neurology; Neurons; Neurosciences; Onset of illness; Pathogenesis; Pathology; Pathway interactions; Phase; Phosphotransferases; Postdoctoral Fellow; Production; Protein Isoforms; Protein Kinase; Proteins; ROCK1 gene; Research; Resistance; Rodent; Scientist; Signal Transduction; Societies; Staging; Students; Symptoms; Synaptic plasticity; Techniques; Testing; Therapeutic; Time; Training; Translating; Universities; Wood material; Work; abeta accumulation; abstracting; aging brain; base; behavior test; brain tissue; career; cognitive function; cognitive reserve; cost; design; end stage disease; extracellular; improved; in vivo; innovation; insight; light microscopy; meetings; member; mouse model; mutant; nervous system disorder; neuropathology; prevent; programs; rho; rho GTP-Binding Proteins; skills; small hairpin RNA; symposium

Project Summary/Abstract Candidate: My career goal is to be a successful, independent academic research scientist with a laboratory that contributes important insights into Alzheimer's disease (AD) as well as other neurodegenerative diseases and fosters the training of students and fellows. My long-term research goals are to translate basic science discoveries about the mechanisms of neurodegeneration and AD pathogenesis into rational therapies. The K99/R00 Pathway to Independence award will facilitate important career training that will provide a path to establish my own independent research group and initiate new avenues of AD research. Training: Dr. James Lah will be my primary mentor and Dr. Allan Levey will be my co-mentor for the K99 phase of this application, and working together we have developed a plan to provide the necessary career training that will allow for the execution of the proposed research and my transition to independence. In addition to my mentors, a committee of faculty members, including Drs. Gary Bassell, David Weinshenker, Marla Gearing, Ranjita Betarbet, and Richard Kahn, will provide intellectual guidance and technical support during the K99 phase. Drs. Bassell will share his expertise with state-of-the-art microscopy techniques, while Dr. Weinshenker will provide training for rodent behavioral testing. Drs. Gearing and Betarbet will assist with immunohistochemical studies of postmortem human brain tissues and stereotaxic injection of mice, respectively, and Dr. Kahn will offer wisdom and career guidance to facilitate my transition to independence. In the first year of the K99 phase, I will attend a Cold Springs Harbor Laboratory training course in Molecular Neurology and Neuropathology and the Analytical & Quantitative Light Microscopy course at the Marine Biological Laboratory in Woods Hole. In year two, I will attend the University of Pittsburgh Course in Scientific Management and Leadership which is an interactive learning forum designed to equip senior postdoctoral fellows with the knowledge and professional competencies to lead innovative and productive research programs. As a postdoctoral fellow, I have presented my research every year at the annual Society for Neuroscience meeting, and I will continue this tradition through the K99 and R00 phases. I also will present my findings at the International Conference on Alzheimer's disease in 2013. I will attend weekly seminars in the Dept. of Neurology and Cell Biology and participate in a training course on scientific ethics sponsored by the Emory University Office of Postdoctoral Education. Research: Cognitive decline is a clinical hallmark of progression from healthy brain aging to Alzheimer's disease (AD), while increased production and accumulation of amyloid-β (Aβ) is a pathological hallmark of AD. There is strong evidence that the observed cognitive impairment in AD is in part due to Aβ's negative impact on synaptic plasticity. Therefore, designing therapeutics that simultaneously boost cognitive reserve and function as well as decrease Aβ production may prevent AD onset and treat end-stage disease. RhoA, a Rho GTPase family member, is a well-studied regulator of neuronal outgrowth and synaptic plasticity. The principle downstream effectors of RhoA are the Rho-associated coiled-coil containing protein kinases (ROCK), ROCK1 and ROCK2. Active RhoA promotes antagonistic effects on synaptic plasticity, and extracellular Aβ induces RhoA activity. Furthermore, there is an accordant relationship between RhoA activity and Aβ generation. Development of AD pathology likely begins many years prior to clinical symptom onset, and during this time, we propose that Aβ accumulation activates the RhoA/ROCK pathway which thereby negatively impacts synaptic plasticity and fuels production of Aβ. The RhoA/ROCK pathway is an exciting target for pharmacologic intervention, but progress is hampered by the ambiguity of which downstream RhoA signaling events are attributable to ROCK1 or ROCK2 in brain. Importantly, the proposed studies will be the first to evaluate targeted disruption of ROCK1 or ROCK2 in the same experimental model system in vivo. We hypothesize that RhoA/ROCK activity increases as AD progresses and that inhibition of the RhoA-ROCK2 pathway will improve cognitive function in AD. To test this postulate, we will determine if RhoA/ROCK activity is amplified in asymptomatic AD (early stages of disease) and symptomatic AD (end-stage disease) cases by measuring RhoA, ROCK1, and ROCK2 activity in control, asymptomatic AD, and symptomatic AD brains. Next, we will determine how targeted knockdown of ROCK1 or ROCK2 affects cognition dysfunction and Aβ deposition in an AD mouse model by delivering adeno-associated virus expressing ROCK isoform specific shRNA to the hippocampus. Finally, we will test the model that activation or inhibition of RhoA reflects ROCK2-mediated effects in brain by measuring changes in dendritic spine morphology as well as Aβ production following pharmacological modulation of RhoA activity coupled with targeted shRNA reduction of ROCK1 or ROCK2. Results from these studies will facilitate future development of drugs targeted against the RhoA/ROCK pathway for the treatment and prevention of AD. In addition, the intellectual and technical skills I acquire over the K99 training period will allow me to establish an independent research program to investigate the functional importance of specific ROCK isoforms in AD as well as other models of neurological disorders.

项目概要/摘要 候选人：我的职业目标是成为一名成功的、拥有实验室的独立学术研究科学家 为了解阿尔茨海默病 (AD) 以及其他神经退行性疾病提供了重要见解 并促进学生和研究员的培训我的长期研究目标是转化基础科学。 将神经退行性变和 AD 发病机制的发现转化为合理的治疗方法。 K99/R00 独立之路奖将促进重要的职业培训，从而提供一条通往独立之路 建立自己的独立研究小组并开创AD研究的新途径。 培训：James Lah 博士将是我的主要导师，Allan Levey 博士将是我 K99 的联合导师 在此申请的阶段，我们共同制定了一项计划，以提供必要的职业 培训将使我能够执行拟议的研究并过渡到独立。 除了我的导师之外，还有一个由教师组成的委员会，包括加里·巴塞尔 (Gary Bassell)、大卫·温申克 (David Weinshenker) 博士、 Marla Gearing、Ranjita Betarbet 和 Richard Kahn 将提供智力指导和技术支持 在 K99 阶段，Bassell 博士将分享他在最先进的显微镜技术方面的专业知识。 Weinshenker 博士将提供啮​​齿动物行为测试培训，而 Betarbet 博士将提供协助。 死后人脑组织和小鼠立体定位注射的免疫组织化学研究， 卡恩博士将分别提供智慧和职业指导，以帮助我过渡到独立。 K99 阶段的第一年，我将参加冷泉港实验室分子培训课程 海军陆战队的神经病学和神经病理学以及分析和定量光学显微镜课程 第二年，我将参加伍兹霍尔的生物实验室，参加匹兹堡大学的科学课程。 管理与领导力，这是一个互动学习论坛，旨在装备高级博士后 具有领导创新和富有成效的研究的知识和专业能力的研究员 作为一名博士后研究员，我每年都会在年度学会上展示我的研究成果。 神经科学会议，我将在 K99 和 R00 阶段延续这一传统，我也将展示我的成果。 2013 年国际阿尔茨海默病会议上的研究结果。我将参加每周举行的研讨会 神经病学和细胞生物学系，并参加由该院主办的科学伦理培训课程 埃默里大学博士后教育办公室。 研究：认知能力下降是从健康大脑老化到阿尔茨海默病进展的临床标志 β 淀粉样蛋白 (Aβ) 的产生和积累增加是 AD 的病理标志。 有强有力的证据表明，AD 中观察到的认知障碍部分是由于 Aβ 的负面影响 因此，同时增强认知储备和突触可塑性的治疗设计。 功能以及减少 Aβ 的产生可以预防 AD 发病并治疗终末期疾病 RhoA。 GTPase 家族成员，是一种经过充分研究的神经元生长和突触可塑性调节剂。 RhoA 的下游效应是含有 Rho 相关卷曲螺旋的蛋白激酶 (ROCK)、ROCK1 ROCK2 促进突触可塑性的拮抗作用，细胞外 Aβ 诱导。 此外，RhoA 活性与 Aβ 生成之间存在一致的关系。 AD 病理学的发展可能在临床症状出现前许多年就开始了，在此期间， 我们认为 Aβ 积累会激活 RhoA/ROCK 通路，从而产生负面影响 RhoA/ROCK 通路是一个令人兴奋的药理学靶点。 干预，但进展受到下游 RhoA 信号事件的模糊性的阻碍 重要的是，拟议的研究将是第一个评估大脑中的 ROCK1 或 ROCK2 的研究。 在同一体内实验模型系统中靶向破坏 ROCK1 或 ROCK2。 RhoA/ROCK 活性随着 AD 的进展而增加，并且 RhoA-ROCK2 通路的抑制将会改善 为了测试这一假设，我们将确定 RhoA/ROCK 活性在 AD 中是否被放大。 通过测量无症状 AD（疾病早期）和有症状 AD（疾病终末期）病例 对照、无症状 AD 和有症状 AD 大脑中的 RhoA、ROCK1 和 ROCK2 活性 接下来，我们将讨论。 确定 ROCK1 或 ROCK2 的靶向敲低如何影响认知功能障碍和 Aβ 沉积 通过将表达 ROCK 亚型特异性 shRNA 的腺相关病毒递送至 AD 小鼠模型 最后，我们将测试 Rho 的激活或抑制反映 ROCK2 介导的模型。 通过测量树突棘形态的变化以及 Aβ 产生后对大脑的影响 RhoA 活性的药理学调节与 ROCK1 或 ROCK2 的靶向 shRNA 减少相结合。 这些研究的结果将有助于未来针对 RhoA/ROCK 的药物的开发 此外，我还获得了治疗和预防 AD 的途径。 K99培训期将使我能够建立一个独立的研究计划来调查功能 特定 ROCK 亚型在 AD 以及其他神经系统疾病模型中的重要性。