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: 8874382
• 负责人: Jeremy H. Herskowitz
• 金额: $ 24.9万
• 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/8874382
• 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.

项目摘要/摘要 应聘者：我的职业目标是成为一名有实验室的成功的、独立的学术研究科学家 这为阿尔茨海默病(AD)和其他神经退行性疾病提供了重要的见解 并促进对学生和研究员的培训。我的长期研究目标是翻译基础科学 神经退行性变和阿尔茨海默病发病机制的发现成为合理的治疗方法。这个 K99/R00独立之路奖将促进重要的职业培训，这将为 建立自己的独立研究小组，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的共同导师 在这个申请阶段，我们共同制定了一个计划，提供必要的职业生涯 培训，这将允许执行拟议的研究和我过渡到独立。在……里面 除了我的导师，还有一个由教职员工组成的委员会，包括加里·巴塞尔博士、大卫·温申克博士、 玛拉·盖林、兰吉塔·贝塔贝特和理查德·卡恩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，而 温申克博士将为啮齿动物行为测试提供培训。吉尔林博士和贝塔贝特博士将协助 死后人脑组织和小鼠立体定向注射的免疫组织化学研究 卡恩博士将提供智慧和职业指导，以促进我向独立的过渡。在……里面 在K99阶段的第一年，我将参加冷泉港实验室的分子培训课程 海军陆战队神经学和神经病理学与分析和定量光学显微镜课程 伍兹霍尔的生物实验室。在第二年，我将参加匹兹堡大学的科学课程 管理和领导力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 程序。作为一名博士后研究员，我每年都会在年度 神经科学会议，我将在K99和R00阶段延续这一传统。我也会向大家展示我的 2013年阿尔茨海默病国际会议上的研究结果。我将参加每周一次的研讨会 部门神经学和细胞生物学教授，并参加由 埃默里大学博士后教育办公室。 研究：认知功能减退是健康脑老化进展为阿尔茨海默氏症的临床标志 阿尔茨海默病(AD)，而淀粉样蛋白(Aβ，Aβ)的产生和积聚是AD的病理标志。 有强有力的证据表明，在AD中观察到的认知障碍部分是由于Aβ的S的负面影响 关于突触的可塑性。因此，设计同时提高认知储备和 功能和减少Aβ的产生可以预防AD的发生和治疗终末期疾病。Rhoa，一个Rho GTP酶家族成员，是神经元生长和突触可塑性的重要调节因子。原则性 RhoA的下游效应物是Rho相关的含有蛋白激酶的卷曲线圈(ROCK)，ROCK1 和ROCK2。活性RhoA促进突触可塑性的拮抗作用，细胞外Aβ诱导 RhoA活动。此外，RhoA活性与Aβ的产生之间存在着一致的关系。 AD病理的发展可能在临床症状出现之前许多年就开始了，在这段时间里， 我们认为，Aβ的积累激活了RhoA/ROCK途径，从而对 突触可塑性与A-β的产生。RhoA/ROCK通路是一个令人兴奋的药理学靶点 干预，但进展受到哪些下游RhoA信号事件的模棱两可的阻碍 可归因于大脑中的ROCK1或ROCK2。重要的是，拟议的研究将是第一个评估 在体内靶向干扰同一实验模型系统中的ROCK1或ROCK2。我们假设 随着AD的进展，RhoA/ROCK活性增加，对RhoA-ROCK2通路的抑制作用将得到改善 阿尔茨海默病的认知功能。为了测试这一假设，我们将确定RhoA/ROCK活动是否在 无症状AD(疾病早期)和有症状AD(终末期疾病)病例 RhoA、ROCK1和ROCK2在对照组、无症状AD和症状性AD脑中的活性。接下来，我们将 确定靶向敲除ROCK1或ROCK2对认知功能障碍和β沉积的影响 将表达ROCK异构体特异性shRNA的腺相关病毒导入AD小鼠模型 海马体。最后，我们将检验RhoA的激活或抑制反映ROCK2介导的模型 脑内树突棘形态的变化和β产生的变化 RhoA活性的药理调节与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 治疗和预防阿尔茨海默病的途径。此外，我多年来学到的智力和技术技能 K99培训期间将允许我建立一个独立的研究计划，以调查功能 特定的岩石亚型在阿尔茨海默病和其他神经性疾病模型中的重要性。