RTP801 and ATF4 in Neuronal Death in PD

RTP801 和 ATF4 在 PD 神经元死亡中的作用

• 批准号: 8510736
• 负责人: Oren Abraham Levy
• 金额: $ 17.43万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8510736
• 关键词: Address; Affect; Alzheimer' s Disease; American; Area; Award; Cell Death; Cell Survival; Cell model; Cessation of life; Clinical; Clinical Research; Clinical Skills; Collaborations; Coupled; Deposition; Development; Disease; Disease model; Doctor of Philosophy; Educational process of instructing; Environment; Equilibrium; Event; Faculty; Fellowship; Foundations; Funding; Genes; Goals; Grant; Hypoxia; In Vitro; K-Series Research Career Programs; Knockout Mice; Lewy Bodies; Link; Location; Manuscripts; Mediating; Medical center; Mentors; Modeling; Molecular Biology Techniques; Movement Disorders; Names; Neurodegenerative Disorders; Neurology; Neurons; Neurosciences; Notch Signaling Pathway; PC12 Cells; Parkinson Disease; Pathogenesis; Pathway interactions; Patient Care; Patients; Phase; Physicians; Play; Proteins; Publishing; RNA Interference; Regulation; Research; Research Personnel; Research Proposals; Residencies; Resources; Role; Scientist; Staging; Stress; Subfamily lentivirinae; Substantia nigra structure; Sum; Syndrome; System; Techniques; Testing; Time; Toxin; Training; Transcriptional Activation; Transcriptional Regulation; Translational Research; Universities; Up-Regulation; Work; alpha synuclein; base; biological adaptation to stress; brain cell; career; cell type; clinical care; disability; disease-causing mutation; drug development; experience; in vivo; insight; interest; member; mouse model; mutant; neuron loss; novel; overexpression; presenilin; public health relevance; research study; response; stressor; success; transcription factor; transduction efficiency

DESCRIPTION (provided by applicant): My goal is to become a physician-scientist investigating basic mechanisms of neurodegenerative disease, specifically Parkinson's disease. I obtained an MD/PhD from Emory University under the direction of Allan Levey. I studied the interaction between the Notch signaling pathway and the Alzheimer's disease-linked presenilin genes, using cellular models and basic molecular biology techniques. During my PhD work, I obtained two grants to fund my work and published two manuscripts. I completed my residency in neurology at the Weill-Cornell Medical Center, where I was named Chief Resident my final year. Next, I came to Columbia for fellowship training in movement disorders. I have developed my clinical skills in caring for patients with movement disorders, primarily Parkinson's disease. During my second year of fellowship (July 2008) I returned to full-time bench research. Since joining the lab I have authored a review article on cell death mechanisms in Parkinson's disease. In sum, I have demonstrated a strong commitment to research throughout all phases of my training, and I believe that I have shown the ability to succeed in carrying out research. I decided to perform my fellowship training at Columbia because it was an ideal location for advancing my career. Columbia's Movement Disorders Division provides excellent clinical training and resources for clinical and translational research. More importantly, Columbia's research environment in neuroscience is outstanding. Specifically, the diverse range of research interests coupled with the high degree of collaboration was very appealing. In addition to its faculty, the University has first-rate facilities, ample institutional funding, and active academic events. When choosing a mentor, I wanted someone who could teach me a novel set of techniques and approaches, but shared my interest in using cellular models to address questions related to disease mechanisms. I chose Lloyd Greene for several reasons: he is a senior faculty member with a strong record of training scientists at my career stage; he is an expert in cell death mechanisms and has an active project in PD-related cell death; he collaborates closely with other researchers in multiple areas of PD research; the techniques and approaches used in the lab are important ones that I have no experience with, e.g. primary neuronal cultures, the use of lentivirus for high efficiency transduction, and RNA interference. My research proposal focuses on Parkinson's disease (PD), a common neurodegenerative disease that causes significant disability. The development of neuroprotective therapies is a pressing need in PD, but this requires a better understanding of the mechanisms of neuronal death in the disease. The Greene lab has studied transcriptionally regulated genes that contribute to either cell death or survival in PD. One of these, a protein called RTP801, appears to be essential for cell death in PD models and is upregulated in affected neurons in the disease. To date, RTP801 has been evaluated in toxin-based models of PD. In order to further validate the role of RTP801 in PD pathogenesis, we will test the role of RTP801 in alpha-synuclein-mediated cell death (Specific Aim 1). The regulation of RTP801 occurs primarily at the transcriptional level, but RTP801 regulation has not been studied in neuronal systems or and in response to stressors relevant to PD. Identifying the transcription factor(s) involved in RTP801 upregulation during stressors relevant to PD may provide potential targets for neuroprotective therapies. Therefore, we will identify the transcription factor(s) important for regulating RTP801 in response to PD-relevant stressors (Specific Aim 2). We have studied the transcription factor ATF4 as a potential regulator of RTP801 and cell death, since ATF4 activity is involved in increasing RTP801 expression in other systems. In preliminary experiments, we found that ATF4 is not required for RTP801 upregulation by 6OHDA. Furthermore, 6OHDA-induced cell death was exacerbated by decreasing ATF4 levels, suggesting that ATF4 may be protective. If so, augmenting ATF4 activity might be a strategy for delaying neuronal cell death in PD. We will further define the role of ATF4 in toxin-mediated cell death and RTP801 upregulation. Furthermore, we plan to test the notion that ATF4 activity is neuroprotective in PD in the 1Syn cellular model mentioned above and the in vivo MPTP model using ATF4-null mice (Specific Aim 3). In conclusion, this career development award is essential for enabling me to become an independent physician-scientist. Having been away from bench research for more than 7 years, I need to re-establish a project and line of research, essentially from scratch. For bench research, the ability to limit my clinical responsibilities and spend the majority of my time in lab is crucial for success. During the period of the award, I will spend 80% of my time performing research; the remainder of the time will be spent in clinical care of PD patients and teaching. Over the course of the grant, I will gain experience with a group of approaches and systems that will allow me to explore fundamental disease mechanisms in PD. I will develop a theme of research regarding the beneficial and harmful effects of stress-related proteins and pathways in PD. As the award draws to a close, I will use the story and findings I have developed as a foundation for obtaining independent research funding.

描述（由申请人提供）：我的目标是成为一名研究神经退行性疾病，特别是帕金森病的基本机制的医生科学家。我在艾伦·利维的指导下获得了埃默里大学的医学博士学位。我研究了Notch信号通路和阿尔茨海默病相关的早老素基因之间的相互作用，使用细胞模型和基本的分子生物学技术。在我攻读博士学位期间，我获得了两赠款，并发表了两篇手稿。我在威尔-康奈尔医疗中心完成了神经病学的住院医师实习，在那里我被任命为住院总医师。接下来，我来到哥伦比亚参加运动障碍的奖学金培训。我在照顾运动障碍患者方面发展了我的临床技能，主要是帕金森病。在我的第二年的奖学金（2008年7月），我回到全职板凳研究。自从加入实验室以来，我撰写了一篇关于帕金森病细胞死亡机制的评论文章。总之，我在培训的各个阶段都表现出了对研究的坚定承诺，我相信我已经表现出了成功开展研究的能力。 我决定在哥伦比亚大学进行奖学金培训，因为这是一个促进我职业发展的理想场所。哥伦比亚的运动障碍部门提供了良好的临床培训和资源的临床和转化研究。更重要的是，哥伦比亚在神经科学方面的研究环境非常出色。具体而言，多样化的研究兴趣加上高度的合作是非常有吸引力的。除了教师，大学还拥有一流的设施，充足的机构资金和活跃的学术活动。 在选择导师时，我希望有人能教我一套新的技术和方法，但与我分享使用细胞模型来解决疾病机制相关问题的兴趣。我选择Lloyd格林有以下几个原因：他是一名资深教员，在我的职业生涯阶段有很好的培训科学家的记录;他是细胞死亡机制方面的专家，在PD相关的细胞死亡方面有一个活跃的项目;他在PD研究的多个领域与其他研究人员密切合作;实验室中使用的技术和方法是我没有经验的重要技术和方法，例如原代神经元培养、使用慢病毒进行高效转导和RNA干扰。 我的研究计划集中在帕金森病（PD），一种常见的神经退行性疾病，导致显着残疾。神经保护疗法的发展是PD的迫切需要，但这需要更好地了解疾病中神经元死亡的机制。格林实验室研究了转录调控基因，这些基因对帕金森病中的细胞死亡或存活有贡献。其中一种称为RTP 801的蛋白质似乎对PD模型中的细胞死亡至关重要，并且在疾病中受影响的神经元中上调。迄今为止，RTP 801已在基于毒素的PD模型中进行了评估。为了进一步验证RTP 801在PD发病机制中的作用，我们将测试RTP 801在α-突触核蛋白介导的细胞死亡中的作用（特异性目的1）。RTP 801的调节主要发生在转录水平，但RTP 801的调节尚未在神经元系统中或响应于与PD相关的应激源进行研究。鉴定在与PD相关的应激期间参与RTP 801上调的转录因子可能为神经保护疗法提供潜在靶点。因此，我们将鉴定对调节RTP 801以响应PD相关应激物重要的转录因子（具体目标2）。我们已经研究了转录因子ATF 4作为RTP 801和细胞死亡的潜在调节因子，因为ATF 4活性参与增加其他系统中的RTP 801表达。在初步实验中，我们发现ATF 4不是RTP 801通过6 OHDA上调所必需的。此外，6 OHDA诱导的细胞死亡通过降低ATF 4水平而加剧，表明ATF 4可能具有保护作用。如果是这样的话，增强ATF 4活性可能是延迟PD中神经元细胞死亡的一种策略。我们将进一步确定ATF 4在毒素介导的细胞死亡和RTP 801上调中的作用。此外，我们计划在上述1 Syn细胞模型和使用ATF 4缺失小鼠的体内MPTP模型中测试ATF 4活性在PD中具有神经保护作用的概念（具体目标3）。 总之，这个职业发展奖是必不可少的，使我成为一个独立的医生，科学家。我已经离开实验室研究7年多了，我需要重新建立一个项目和研究路线，基本上是从头开始。对于台架研究，限制我的临床责任并将大部分时间花在实验室的能力对于成功至关重要。在获奖期间，我将花费80%的时间进行研究;其余时间将用于PD患者的临床护理和教学。在资助的过程中，我将获得一组方法和系统的经验，这将使我能够探索PD的基本疾病机制。我将开发一个主题的研究有关的有益和有害的影响压力相关的蛋白质和途径在PD。随着奖项接近尾声，我将利用我所开发的故事和发现作为获得独立研究资金的基础。