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.

项目总结/摘要 候选人：我的职业目标是成为一名成功的、独立的学术研究科学家， 这有助于对阿尔茨海默病（AD）以及其他神经退行性疾病的重要见解 并促进学生和研究员的培训。我的长期研究目标是将基础科学 关于神经变性和AD发病机制的发现转化为合理的治疗方法。的 K99/R 00独立之路奖将促进重要的职业培训，这将提供一条通往 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博士将是我在K99上的共同导师 这个应用程序的阶段，并共同努力，我们已经制定了一个计划，以提供必要的职业生涯 培训，这将允许执行拟议的研究和我的过渡到独立。在 除了我的导师，还有一个由教师组成的委员会，包括加里·巴塞尔博士，大卫·温申克博士， 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， 博士Weinshenker将提供啮齿动物行为测试的培训。吉尔林医生和贝塔贝特医生会协助 死后人脑组织的免疫组织化学研究和小鼠的立体定位注射， 卡恩博士将提供智慧和职业指导，以促进我向独立过渡。在 在K99阶段的第一年，我将参加冷泉港实验室的分子生物学培训课程， 神经病学和神经病理学以及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， 管理与领导力是一个互动学习论坛，旨在为高级博士后提供知识 具有知识和专业能力的研究员，以领导创新和富有成效的研究 程序.作为一名博士后研究员，我每年都在年度社会上展示我的研究成果。 神经科学会议上，我将继续通过K99和R 00阶段这个传统。我也将介绍我的 2013年阿尔茨海默病国际会议上的发现。我将参加每周的研讨会， 部并参加了由国际神经病学和细胞生物学学会主办的科学道德培训班。 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氏症进展的临床标志 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归因于Aβ的负面影响 关于突触可塑性因此，设计同时提高认知储备和 功能以及减少Aβ的产生可以预防AD发作和治疗终末期疾病。RhoA，a Rho GT3家族成员，是神经元生长和突触可塑性的充分研究的调节剂。原则 RhoA的下游效应物是Rho相关的含有卷曲螺旋的蛋白激酶（ROCK），ROCK 1 摇滚2活性RhoA促进对突触可塑性的拮抗作用，细胞外Aβ诱导 RhoA活性。此外，RhoA活性与Aβ生成之间存在着一致的关系。 AD病理学的发展可能在临床症状发作前多年开始，并且在此期间， 我们认为，Aβ积累激活了RhoA/ROCK通路，从而对 突触可塑性和Aβ的产生。RhoA/ROCK通路是药理学研究的一个令人兴奋的靶点。 干预，但进展受到下游RhoA信号传导事件的模糊性的阻碍 ROCK 1或ROCK 2在大脑中的作用。重要的是，拟议的研究将是第一个评估 在相同的体内实验模型系统中靶向破坏ROCK 1或ROCK 2。我们假设 RhoA/ROCK活性随着AD的进展而增加，并且RhoA-ROCK 2通路的抑制将改善AD的治疗效果。 AD的认知功能为了验证这一假设，我们将确定RhoA/ROCK活性是否在细胞中扩增。 无症状AD（疾病早期）和有症状AD（终末期疾病）病例， RhoA，ROCK 1和ROCK 2在对照，无症状AD和有症状AD大脑中的活性。接下来我们就 确定ROCK 1或ROCK 2的靶向敲除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 海马体。最后，我们将测试RhoA的激活或抑制反映ROCK 2介导的细胞凋亡的模型。 通过测量树突棘形态学的变化以及Aβ的产生， RhoA活性的药理学调节与ROCK 1或ROCK 2的靶向shRNA减少偶联。 这些研究的结果将有助于未来开发针对RhoA/ROCK的药物 治疗和预防AD的途径。此外，我所获得的知识和技术技能， K99培训期间将使我能够建立一个独立的研究计划，以调查功能 在AD以及其他神经系统疾病模型中特定ROCK亚型的重要性。