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: 8549081
• 负责人: Jeremy H. Herskowitz
• 金额: $ 8.56万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-30 至 2014-04-11
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8549081
• 关键词: Address; Affect; Alzheimer disease prevention; Alzheimer' s Disease; American; Amino Acid Substitution; Amyloid; Amyloid deposition; Autopsy; Award; Basic Science; Binding; Biological; Biological Models; Brain; Cells; Cellular biology; Clinical; Cognition; Coupled; Data; Dementia; Dendritic Spines; Dependovirus; 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; 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

DESCRIPTION (provided by applicant): 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. Dr. 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-¿ (A¿) is a pathological hallmark of AD. There is strong evidence that the observed cognitive impairment in AD is in part due to A¿'s negative impact on synaptic plasticity. Therefore, designing therapeutics that simultaneously boost cognitive reserve and function as well as decrease A¿ 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 A¿ induces RhoA activity. Furthermore, there is an accordant relationship between RhoA activity and A¿ generation. Development of AD pathology likely begins many years prior to clinical symptom onset, and during this time, we propose that A¿ accumulation activates the RhoA/ROCK pathway which thereby negatively impacts synaptic plasticity and fuels production of A¿. 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 A¿ 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 A¿ 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.

职位描述(申请人提供)：我的职业目标是成为一名成功的、独立的学术研究科学家，拥有一个实验室，对阿尔茨海默氏症(AD)和其他神经退行性疾病提供重要见解，并促进学生和研究员的培训。我的长期研究目标是将关于神经退行性变和AD发病机制的基础科学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，这将为我建立自己的独立研究小组和启动AD研究的新途径提供一条途径。培训：James Lah博士将是我的主要导师，Allan Levey博士将是我在K99申请阶段的共同导师，我们共同制定了一项计划，提供必要的职业培训，以便执行拟议的研究并过渡到独立。除了我的导师，一个由加里·巴塞尔博士、大卫·温申克博士、玛拉·盖林博士、兰吉塔·贝塔贝特博士和理查德·卡恩博士组成的教职员工委员会将在K99阶段提供智力指导和技术支持。巴塞尔博士将分享他在最先进的显微镜技术方面的专业知识，而温申克博士将提供啮齿动物行为测试方面的培训。盖林博士和贝塔贝特博士将分别协助对死后人脑组织的免疫组织化学研究和对小鼠进行立体定向注射，卡恩博士将提供智慧和职业指导，帮助我过渡到独立。在K99阶段的第一年，我将参加冷泉港实验室分子神经学和神经病理学的培训课程，以及伍兹霍尔海洋生物实验室的分析和定量光学显微镜课程。在第二年，我将参加匹兹堡大学的科学管理和领导力课程，这是一个互动学习论坛，旨在让高级博士后研究员掌握领导创新和富有成效的研究项目的知识和专业能力。作为一名博士后研究员，我每年都会在神经科学学会的年度会议上展示我的研究成果，我将在K99和R00阶段延续这一传统。我还将在2013年的阿尔茨海默病国际会议上介绍我的发现。我将参加每周一次的系内研讨会。并参加由埃默里大学博士后教育办公室主办的科学伦理培训课程。研究：认知衰退是从健康的大脑老化到阿尔茨海默病(AD)的临床标志，而淀粉样蛋白(A？)的产生和积累增加是AD的病理标志。有强有力的证据表明，观察到的AD认知障碍部分是由于S对突触可塑性的负面影响。因此，设计同时提高认知储备和功能，同时减少Aβ产生的治疗方法，可能会预防AD的发病和治疗终末期疾病。RhoA是Rho GTP酶家族成员之一，是神经元生长和突触可塑性的重要调节因子。RhoA的主要下游效应子是Rho相关的含有蛋白激酶(ROCK)的螺旋线圈，即ROCK1和ROCK2。活性RhoA促进对突触可塑性的拮抗作用，而细胞外A则诱导RhoA活性。此外，RhoA活性与A？代之间存在着一致的关系。AD的病理发展可能在临床症状出现之前许多年就开始了，在此期间，我们认为A？的积聚激活了RhoA/ROCK途径，从而对突触的可塑性产生了负面影响，并促进了A？的产生。RhoA/ROCK通路是药物干预的一个令人兴奋的靶点，但由于RhoA下游信号事件与脑内ROCK1或ROCK2的关系不明确，这一研究进展受到了阻碍。重要的是，拟议的研究将是第一次在体内评估在同一实验模型系统中对ROCK1或ROCK2的靶向干扰。我们假设RhoA/ROCK活性随着AD的进展而增加，抑制RhoA-ROCK2通路将改善AD的认知功能。为了验证这一假设，我们将通过测量对照组、无症状AD和有症状AD脑中的RhoA、ROCK1和ROCK2活性，来确定RhoA/ROCK活性在无症状AD(疾病的早期阶段)和有症状AD(终末期疾病)病例中是否被放大。接下来，我们将通过将表达ROCK亚型特异性shRNA的腺相关病毒传递到海马区，确定靶向敲除ROCK1或ROCK2如何影响AD小鼠模型中的认知功能障碍和A？沉积。最后，我们将通过测量药物调节RhoA活性并靶向减少ROCK1或ROCK2的shRNA后树突棘形态和A？产生的变化来测试RhoA的激活或抑制反映ROCK2在脑中的作用的模型。这些研究的结果将有助于未来针对RhoA/ROCK途径的药物的开发，用于治疗和预防AD。此外，我在K99培训期间获得的智力和技术技能将使我能够建立 一个独立的研究计划，调查特定的岩石异构体在AD和其他神经性疾病模型中的功能重要性。