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: 8097526
• 负责人: Oren Abraham Levy
• 金额: $ 17.2万
• 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/8097526
• 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. PUBLIC HEALTH RELEVANCE: Parkinson's disease affects over 1 million Americans and causes significant disability. This project aims to identify proteins that contribute to the death of brain cells in this disease, with the goal of identifying new targets for drug development.

描述(申请人提供)：我的目标是成为一名内科科学家，研究神经退行性疾病的基本机制，特别是帕金森氏症。在艾伦·利维的指导下，我获得了埃默里大学的医学博士学位。我使用细胞模型和基本的分子生物学技术，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。在我的博士工作期间，我获得了两笔资金来资助我的工作，并发表了两篇手稿。我在威尔-康奈尔医学中心完成了神经学住院医师的实习，在那里我被任命为首席住院医师。接下来，我来到哥伦比亚接受运动障碍方面的团契培训。我在护理运动障碍患者方面发展了自己的临床技能，主要是帕金森氏症。在我获得奖学金的第二年(2008年7月)，我回到了全职的板凳研究。自从加入实验室以来，我撰写了一篇关于帕金森氏症细胞死亡机制的评论文章。总而言之，在我的培训的所有阶段，我都表现出了对研究的强烈承诺，我相信我已经展示了成功开展研究的能力。我决定在哥伦比亚大学接受奖学金培训，因为那里是我职业发展的理想地点。哥伦比亚大学的运动障碍分部为临床和转化性研究提供出色的临床培训和资源。更重要的是，哥伦比亚大学在神经科学方面的研究环境非常出色。具体地说，多样化的研究兴趣和高度的合作非常吸引人。除了教职员工外，该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，我想要一个可以教我一套新的技术和方法的人，但也和我一样，对使用细胞模型来解决与疾病机制相关的问题感兴趣。我选择Lloyd Greene有几个原因：他是一名资深教员，在我的职业生涯阶段培训科学家方面有很强的记录；他是细胞死亡机制方面的专家，并在PD相关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。我的研究建议集中在帕金森氏病(PD)上，这是一种常见的神经退行性疾病，会导致严重的残疾。发展神经保护疗法是帕金森病的迫切需要，但这需要更好地了解该病中神经元死亡的机制。格林实验室研究了与帕金森病细胞死亡或存活有关的转录调控基因。其中一种名为RTP801的蛋白质似乎是帕金森病模型中细胞死亡的关键，并在疾病中受影响的神经元中上调。到目前为止，RTP801已经在基于毒素的帕金森病模型中进行了评估。为了进一步验证RTP801在帕金森病发病机制中的作用，我们将测试RTP801在α-突触核蛋白介导的细胞死亡中的作用(特异性目标1)。RTP801的调控主要发生在转录水平，但RTP801的调控尚未在神经系统或与帕金森病相关的应激源的反应中进行研究。识别在与帕金森病相关的应激源中参与RTP801上调的转录因子(S)可能为神经保护治疗提供潜在的靶点。因此，我们将确定在调节RTP801对PD相关应激源的反应中起重要作用的转录因子(S)(特定目标2)。我们已经研究了转录因子ATF4作为RTP801和细胞死亡的潜在调节因子，因为ATF4的活性参与了在其他系统中增加RTP801的表达。在初步实验中，我们发现ATF4不是60HDA上调RTP801所必需的。此外，ATF4水平的降低加剧了6OHDA诱导的细胞死亡，提示ATF4可能具有保护作用。如果是这样的话，增强ATF4活性可能是延缓帕金森病神经细胞死亡的一种策略。我们将进一步确定ATF4在毒素介导的细胞死亡和RTP801上调中的作用。此外，我们计划在上述1Syn细胞模型和使用ATF4缺失小鼠的体内MPTP模型(特定目标3)中测试ATF4活性在帕金森病中具有神经保护作用的概念。总而言之，这个职业发展奖对于我成为一名独立的内科科学家是必不可少的。在离开板凳研究7年多之后，我需要重新建立一个项目和研究路线，基本上是从头开始。对于长凳研究来说，限制我的临床责任并将大部分时间花在实验室的能力是成功的关键。在获奖期间，我将花费80%的时间进行研究；其余的时间将用于PD患者的临床护理和教学。在赠款过程中，我将获得一组方法和系统的经验，这些方法和系统将使我能够探索PD的基本疾病机制。我将开发一个关于压力相关蛋白和通路在帕金森病中的有益和有害影响的研究主题。随着奖项接近尾声，我将利用我开发的故事和发现作为获得独立研究资金的基础。 公共卫生相关性：帕金森氏症影响了100多万美国人，并导致严重残疾。该项目旨在确定导致这种疾病中脑细胞死亡的蛋白质，目的是确定药物开发的新靶点。