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（由申请人提供）：我的职业目标是成为一名成功的独立学术研究科学家，拥有一个实验室，为阿尔茨海默病 (AD) 以及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。学期研究目标是将有关神经退行性疾病和 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提供一条道路。培训：James Lah 博士将是我的主要导师，Allan Levey 博士将是我在本申请 K99 阶段的共同导师，我们共同制定了一项计划，以提供除了我的导师之外，还有一个由教师组成的委员会，包括加里·巴塞尔博士、大卫·温申克、玛拉·吉尔林、兰吉塔·贝塔贝特，和 Richard Kahn 博士将在 K99 阶段提供智力指导和技术支持。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 Weinshenker 博士将提供啮​​齿动物行为测试的培训。 Betarbet 将分别协助死后人类脑组织的免疫组织化学研究和小鼠的立体定位注射，而 Kahn 博士将提供智慧和职业指导，以促进我在第一年向独立过渡。 K99 阶段，我将参加冷泉港实验室的分子神经学和神经病理学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。第二年，我将参加匹兹堡大学的科学管理课程。领导力是一个互动学习论坛，旨在为高级博士后研究员提供领导创新和富有成效的研究项目的知识和专业能力，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 阶段继续这一传统，我还将在 2013 年阿尔茨海默病国际会议上展示我的发现。我将参加神经病学和细胞生物学系的每周研讨会，并参加由埃默里大学博士后教育办公室赞助的科学伦理培训课程：认知能力下降是从健康大脑衰老到阿尔茨海默病（AD）的临床标志，同时认知能力的产生和积累增加。淀粉样蛋白-¿ (A¿) 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¿因此，设计能够同时增强认知储备和功能并降低 A¿ 的疗法。 RhoA 是 Rho GTP 酶家族成员，是一种经过充分研究的神经元生长和突触可塑性调节因子，其主要下游效应器是含有 Rho 相关卷曲螺旋的蛋白激酶。 (ROCK)、ROCK1 和 ROCK2 促进对突触可塑性和细胞外 A¿诱导RhoA活性此外，RhoA活性和A¿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，我们建议 A¿积累会激活 RhoA/ROCK 通路，从而对突触可塑性产生负面影响并促进 A¿ RhoA/ROCK 通路是药物干预的一个令人兴奋的目标，但由于大脑中哪些下游 RhoA 信号传导事件归因于 ROCK1 或 ROCK2 的模糊性，进展受到阻碍。重要的是，拟议的研究将是第一个评估 ROCK1 的靶向破坏的研究。我们在同一体内实验模型系统中发现，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，我们将通过测量对照中的 RhoA、ROCK1 和 ROCK2 活性来确定 RhoA/ROCK 活性是否在无症状 AD（疾病早期）和有症状 AD（疾病末期）病例中增强。接下来，我们将确定 ROCK1 或 ROCK2 的靶向敲除如何影响认知功能障碍和 A¿最后，我们将通过测量树突棘形态的变化来测试 RhoA 的激活或抑制反映了 ROCK2 介导的大脑效应的模型。作为A?? RhoA 活性的药理调节与 ROCK1 或 ROCK2 的靶向 shRNA 减少相结合，将有助于未来开发针对 RhoA/ROCK 途径的药物，用于治疗和预防 AD。我在 K99 培训期间获得的知识将使我能够建立 一项独立研究计划，旨在研究特定 ROCK 异构体在 AD 以及其他神经系统疾病模型中的功能重要性。 公共健康相关性：阿尔茨海默病是痴呆的主要原因，目前尚无已知的治疗方法，该通路影响阿尔茨海默病发病机制的关键方面，包括认知能力下降和淀粉样蛋白积累。 ROCK通路将促进针对该通路的药物的开发，用于治疗和预防阿尔茨海默病。