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RTP801 and ATF4 in Neuronal Death in PD

RTP801 和 ATF4 在 PD 神经元死亡中的作用

基本信息

批准号：
8696892
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
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-01 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8696892
关键词：
Address Affect Alzheimer&apos 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 research study response stressor success transcription factor transduction efficiency

项目摘要

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，特别是帕金森病基本机制的内科科学家。我在埃默里大学获得了医学博士学位，师从艾伦·利维。我研究了Notch信号通路与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。在我的博士工作期间，我获得了两项资助来资助我的工作，并发表了两篇手稿。我在威尔-康奈尔医学中心完成了神经病学实习，最后一年被任命为总住院医师。接下来，我来到哥伦比亚大学接受运动障碍方面的奖学金培训。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（2008年7月），我回到全职的实验研究。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，在我接受培训的各个阶段，我都表现出了对研究的强烈承诺，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，因为它是我职业发展的理想场所。哥伦比亚大学运动障碍科为临床和转化研究提供优秀的临床培训和资源。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。具体来说，多样化的研究兴趣加上高度的合作非常有吸引力。除了师资力量外，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，我希望他能教我一套新颖的技术和方法，但在使用细胞模型来解决与疾病机制相关的问题方面与我有共同的兴趣。我选择劳埃德·格林有几个原因：他是一名资深教员，在我的职业生涯阶段培养科学家的记录很好；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都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，以及RNA干扰。我的研究方向是帕金森病，这是一种常见的神经退行性疾病，会导致严重的残疾。神经保护疗法的发展是帕金森病的迫切需要，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PD患者的细胞死亡或存活都有影响。其中一种名为RTP801的蛋白质似乎对PD模型中的细胞死亡至关重要，并且在该疾病中受影响的神经元中表达上调。迄今为止，RTP801已在基于毒素的PD模型中进行了评估。为了进一步验证RTP801在PD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（Specific Aim 1）。RTP801的调控主要发生在转录水平，但尚未研究RTP801在神经元系统中的调控或对PD相关应激源的反应。确定与PD相关的应激源中参与RTP801上调的转录因子可能为神经保护治疗提供潜在的靶点。因此，我们将确定在pd相关应激源下调控RTP801的重要转录因子（Specific Aim 2）。我们研究了转录因子ATF4作为RTP801和细胞死亡的潜在调节因子，因为ATF4的活性参与了其他系统中RTP801表达的增加。在前期实验中，我们发现6OHDA上调RTP801并不需要ATF4。此外，6ohda诱导的细胞死亡随着ATF4水平的降低而加剧，表明ATF4可能具有保护作用。如果是这样，增加ATF4的活性可能是延迟PD中神经元细胞死亡的策略。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，我们计划在上述的1Syn细胞模型和体内MPTP模型中使用ATF4缺失的小鼠来验证ATF4活性对PD的神经保护作用（Specific Aim 3）。综上所述，这个职业发展奖项对我成为一名独立的内科科学家至关重要。已经远离板凳研究超过7年，我需要重新建立一个项目和研究线，基本上从零开始。对于实验室研究，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。在获得资助的过程中，我将获得一系列方法和系统的经验，这些方法和系统将使我能够探索帕金森病的基本疾病机制。我将发展一个关于应激相关蛋白和途径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，我将利用我的故事和发现作为获得独立研究资金的基础。