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: 8425721
• 负责人: Jeremy H. Herskowitz
• 金额: $ 8.88万
• 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 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8425721
• 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 Delivery Systems; 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. PUBLIC HEALTH RELEVANCE: There are no known therapies for the underlying disease-causing mechanisms of Alzheimer's disease, the leading cause of dementia. The RhoA/ROCK pathway influences critical aspects of Alzheimer's disease pathogenesis, including cognitive decline and amyloid accumulation. Studying the RhoA/ROCK pathway will facilitate the development of drugs targeted against this pathway for the treatment and prevention of Alzheimer's disease.

描述（由适用提供）：我的职业目标是成为一名成功的独立学术研究科学家，其实验室对阿尔茨海默氏病（AD）以及其他神经退行性疾病有重要见解，并促进对学生和研究员的培训。我的长期研究目标是将有关神经退行性和AD发病机理机制的基础科学发现转化为理性疗法。 K99/R00独立奖奖将促进重要的职业培训，这将为建立我自己的独立研究小组并启动广告研究的新途径提供一条途径。培训：詹姆斯·拉赫（James Lah）博士将是我的主要心理，艾伦·莱维（Allan Levey）博士将成为我在本申请的K99阶段的同事，我们共同制定了一项计划，以提供必要的职业培训，以允许执行拟议的研究和我的独立过渡。除了我的导师外，包括博士在内的教职员工委员会。 Gary Bassell，David Weinshenker，Marla Gearing，Ranjita Betarbet和Richard Kahn将在K99阶段提供直通的指导和技术支持。 Bassell博士将通过最先进的显微镜技术分享他的专业知识，而Weinshenker博士将提供啮​​齿动物行为测试的培训。 Gearing和Betarbet博士将分别协助对死后人脑组织和立体定位注射小鼠的免疫组织化学研究，而Kahn博士将提供智慧和职业指导，以促进我过渡到独立性。在K99阶段的第一年，我将参加伍兹霍尔（Woods Hole）海洋生物学实验室的分子神经病学和神经病理学以及分析和定量光学显微镜课程的冷泉港实验室培训课程。在第二年，我将参加匹兹堡大学科学管理和领导力课程，这是一个互动学习论坛，旨在为高级博士后研究员提供知识和专业能力，以领导创新和生产性研究计划。作为博士后研究员，我每年都会在年度神经科学会议上介绍我的研究，我将通过K99和R00阶段继续这一传统。我还将在2013年的国际阿尔茨海默氏病国际会议上介绍我的发现。我将参加神经病学和细胞生物学系的每周一次，并参加由埃默里博士后教育埃默里大学（Emory University）主办的科学伦理学培训课程。研究：认知能力下降是从健康脑老化到阿尔茨海默氏病（AD）的临床标志，而淀粉样蛋白 - 淀粉样蛋白的产生和积累的增加是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A检测对合成可塑性的负面影响。因此，设计仅增强认知储备和功能以及减少A生产的疗法可能会阻止AD发作和治疗终期疾病。 Rho GTPase家族成员Rhoa是神经元生长和合成可塑性的良好调节剂。 RhoA的下游原理是含有Rho相关的盘绕蛋白激酶（Rock），Rock1和Rock2。主动RhoA促进对合成可塑性的拮抗作用，细胞外A诱导RhoA活性。此外，Rhoa活动与A e ostry之间存在着一致的关系。 AD病理学的发展可能始于临床症状发作之前的许多年，在此期间，我们建议A积累激活Rhoa/Rock途径，从而对合成可塑性产生负面影响并燃料的产生a。 Rhoa/Rock Pathway是药物干预的令人兴奋的目标，但是由于脑中下游RhoA信号事件的歧义而阻碍了进步，这归因于大脑中的Rock1或Rock2。重要的是，拟议的研究将是第一个评估在体内同一实验模型系统中靶向破坏Rock1或Rock2的研究。我们假设RhoA/岩石活性随着AD的发展而增加，并且对Rhoa-Rock2途径的抑制作用将改善AD的认知功能。为了测试这一假设，我们将通过测量RhoA，Rock1和Rock2活性在对照，不对称AD和症状AD大脑中测量RhoA/Rock活性是否在不对称AD（疾病的早期）和有症状的AD（末期疾病）病例中扩展。接下来，我们将通过将与腺相关的病毒表达岩石同工型特异性shRNA的靶向敲低如何影响AD小鼠模型中的认知功能障碍和A的沉积。最后，我们将测试该模型，即RhoA的激活或抑制通过测量树突状脊柱形态的变化以及在RhoA活性的药物调制后以及靶向shrna降低岩石或岩石2的药物调制后，反映了大脑中的岩石2介导的作用。这些研究的结果将促进针对RhoA/Rock途径的未来开发用于治疗和预防AD的药物。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，研究了特定岩石同工型在AD以及其他神经系统疾病模型中的功能重要性。 公共卫生相关性：对于阿尔茨海默氏病的基本引起疾病的基本机制，痴呆症的主要原因没有已知的疗法。 Rhoa/Rock途径影响阿尔茨海默氏病发病机理的关键方面，包括认知能力下降和淀粉样蛋白的积累。研究Rhoa/Rock途径将有助于针对这种治疗和预防阿尔茨海默氏病的途径的药物开发。