Cerebrospinal fluid proteome mediated signaling in the developing CNS

发育中的中枢神经系统中脑脊液蛋白质组介导的信号传导

基本信息

批准号：
8599494
负责人：
MARIA LEHTINEN
金额：
$ 24.06万
依托单位：
BOSTON CHILDREN'S HOSPITAL
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：
2010-09-30 至 2014-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8599494
关键词：
Adult Affect Age Aging Area Arts Award Bathing Biochemical Genetics Brain Brain region Breathing Caenorhabditis elegans Cell Proliferation Cell Survival Cells Cerebrospinal Fluid Cerebrum Cessation of life Cilia Commit Complement Cues Cyclic AMP Data Development Developmental Biology Diagnostic Disease Doctor of Philosophy Electroporation Embryo Environment Erinaceidae Experimental Models Feedback Finland Foundations Future Genes Genetic Glial Fibrillary Acidic Protein Goals Growth Growth Factor Health Homeostasis Human Hypothyroidism Immune Sera In Vitro Institutes Insulin-Like Growth Factor II Joints Journals Laboratories Laboratory Research Learning Life Location Mass Spectrum Analysis Mediating Mentors Molecular Mus Myoclonic Epilepsies Nerve Degeneration Neuroepithelial Neuroglia Neurologic Neurons Neurosciences Oxidation-Reduction Pattern Pennsylvania Phase Play Positioning Attribute Postdoctoral Fellow Process Proliferating Proteins Proteome Regulation Research Role Schools Scientist Signal Pathway Signal Transduction Signaling Molecule Solid Source Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Stem cells Structure of choroid plexus Supplementation Surface System Techniques Testing Therapeutic Time Training Transgenic Mice Tretinoin Universities Validation Ventricular abstracting career cell behavior developmental neurobiology experience improved in utero in vivo interest meetings mouse model nerve stem cell nervous system disorder neuroepithelium neurogenesis neuroregulation novel diagnostics novel therapeutic intervention progenitor programs receptor research and development research study skills stem stem cell niche undergraduate education undergraduate research

项目摘要

Project Summary/Abstract My interest in diseases of the nervous system stems from my undergraduate education at the University of Pennsylvania, where as a Wistar Institute undergraduate fellow with Dr. Art McMorris, I investigated extrinsic cues triggering cAMP signaling in glia. As an undergraduate research fellow at Rockefeller University in Dr. Bruce McEwen's laboratory, I investigated how hypothyroidism altered signaling molecules in the brain. These experiences led me to graduate school at Harvard University, where I trained with Dr. Azad Bonni in molecular neuroscience and elucidated signaling pathways that mediate survival and death signaling in mammalian neurons and in vivo in C. elegans. Upon completion of my PhD, I carried out my early postdoctocal training with Dr. Anna-Elina Lehesjoki at the Folkh¿lsan Institute of Genetics in Helsinki, Finland, where I discovered that impaired redox homeostasis is a key mechanism triggering neurodegeneration in the progresive myoclonus epilepsy, Unverricht-Lundborg disease (EPM1). From these research experiences, I have gained experience in molecular neuroscience and its applications to neurologic disease. I joined Dr. Chris Walsh's lab for my second postdoc, in hopes that comprehensive training in developmental neurobiology during the mentored phase of this award would complement my training in molecular neuroscience, and provide unique perspective on the mechanisms underlying neurologic disease throughout life. Dr. Walsh's is a leader in the field of cerebral cortical development research. I will learn fundamental techniques in the lab, ranging from quantitative histological analyses to in utero electroporations. Dr. Walsh has assembled a highly talented group of scientists, which will continue to provide a unique source of support and inspiration in informal conversations and weekly lab meetings and journal clubs. The Walsh lab is located in the Division of Genetics, which also hosts joint weekly data talks with the Division of Neuroscience, to promote collaborative exchanges and assistance with technical and theoretical isues. This intellectually engaging environment will provide an excellent source of discussion and feedback during the mentored phase of the award. Importantly, the Division of Genetics is committed to supporting the transition of postdocs to independent research positions. To this end, I will attend a number of career training seminars and meetings geared at preparing me for launching my independent research career. The long-term goal of my research is to elucidate how the cerebrospinal fluid (CSF) functions as a signaling niche that coordinates a rich interaction of signaling factors, acting at long distances to regulate target cell behavior during health and disease. During cerebral cortical development, rapid changes in proliferating progenitor cells occur almost synchronously across vast areas of the neuroepithelium. The protracted location of neuroepithelial cilia in the CSF suggests a potential role for the CSF as a source of extrinsic signals guiding progenitor proliferation. Since the CSF turns over several times per day, the CSF-choroid-plexus system is ideally suited for triggering rapid and spatially synchronized changes in molecular signaling across large distances. My immediate research goal in this proposal is to investigate the role of embryonic CSF proteome in regulating cortical progenitor proliferation during development. The experiments in Aim1 will use heterochronic explants and cultured stem cells to test the ability of CSF to support the survival, growth, and proliferation of cortical progenitor cells. Since the CSF contains hundreds of proteins, I will then use biochemical and genetic approaches to examine how Igf2 (Insulin-like growth factor 2), a candidate factor identified in our preliminary mass spectrometry analyses, may be actively distributed by the CSF to influence progenitor proliferation (Aim 2). The mentored phase of the award (Aims 1&2) will help refine the experimental models, techniques, and professional skills needed to launch a successful, independent research program examining the mechanisms by which CSF-distributed factors influence target cells in health and disease. The fundamental techniques including neurosphere cultures, quantitative histological analyses, and in utero electroporations, all learned during Aims 1&2, wil prepare me for the experiments proposed in Aim 3, which will explore the roles and mechanisms by which other CSF-borne signaling factors, such as retinoic acid and Sonic hedgehog, are regulated by the CSF to act on target cells at the ventricular surface. By the end of this award period, my independent research laboratory wil have pioneered a new concept in developmental biology in which the CSF plays an active role in cerebral cortical development. The proposed experiments represent a solid foundation for future studies investigating changes in the CSF stem cell niche in aging and age-associated neurologic disease. Since the CSF is a surgically accessible medium in humans and mouse models, the proposed experiments will pave the way towards development of powerful diagnostic and therapeutic approaches.

项目概要/摘要我对神经系统疾病的兴趣源于我在大学的本科教育宾夕法尼亚州，作为威斯塔研究所的本科生，我和 Art McMorris 博士一起研究了外在因素触发神经胶质细胞中 cAMP 信号传导的线索，作为洛克菲勒大学的本科研究员。在布鲁斯·麦克尤恩的实验室中，我研究了甲状腺功能减退症如何改变大脑中的信号分子。经历让我进入了哈佛大学研究生院，在那里我接受了 Azad Bonni 博士的分子生物学培训。神经科学并阐明了介导哺乳动物生存和死亡信号的信号通路完成博士学位后，我进行了早期的博士后培训。与 Anna-Elina Lehesjoki 博士在 Folkh¿芬兰赫尔辛基的 lsan 遗传学研究所，我在那里发现了氧化还原稳态受损是引发进行性神经退行性变的关键机制从这些研究经验中，我获得了肌阵挛癫痫、Unverricht-Lundborg 病（EPM1）。我加入了 Chris Walsh 博士的实验室，积累了分子神经科学及其在神经系统疾病中的应用方面的经验。对于我的第二个博士后，希望在发育神经生物学方面进行全面的培训该奖项的指导阶段将补充我在分子神经科学方面的培训，并提供独特的对整个生命过程中神经系统疾病潜在机制的看法。沃尔什博士是大脑皮层发育研究领域的领军人物，我将学习基础知识。沃尔什博士拥有从定量组织学分析到子宫内电穿孔的实验室技术。聚集了一批才华横溢的科学家，他们将继续提供独特的支持和资源来源沃尔什实验室位于非正式对话和每周实验室会议和期刊俱乐部中的灵感。遗传学部门还与神经科学部门每周举办联合数据会谈，以促进就技术和理论问题进行协作交流和协助。环境将在指导阶段提供良好的讨论和反馈来源重要的是，遗传学部门致力于支持博士后向博士后的过渡。为此，我会参加一些职业培训研讨会和会议。旨在帮助我为开展独立研究生涯做好准备。我研究的长期目标是阐明脑脊液（CSF）如何发挥作用协调信号因子丰富相互作用的信号利基，远距离作用以调节目标健康和疾病期间的细胞行为在大脑皮层发育过程中，增殖的快速变化。祖细胞几乎同时出现在神经上皮的大部分区域。脑脊液中神经上皮纤毛的存在表明脑脊液作为外在信号引导源的潜在作用由于脑脊液每天翻转数次，因此脑脊液-脉络丛系统是非常适合触发大范围内分子信号的快速和空间同步变化我在本提案中的近期研究目标是研究胚胎脑脊液蛋白质组在在发育过程中调节皮质祖细胞增殖。 Aim1中的实验将使用异时外植体和培养的干细胞来测试CSF的能力支持皮质祖细胞的存活、生长和增殖，因为脑脊液含有数百个。然后，我将使用生化和遗传学方法来检查 Igf2（胰岛素样生长因子）如何 2），我们初步质谱分析中确定的候选因子，可能通过以下方式主动分布： CSF 影响祖细胞增殖（目标 2）。该奖项的指导阶段（目标 1 和 2）将有所帮助。完善启动成功所需的实验模型、技术和专业技能，独立研究计划，研究 CSF 分布因素影响目标的机制细胞在健康和疾病中的基本技术，包括神经球培养、定量。组织学分析和子宫内电穿孔，所有这些都是在目标 1 和 2 中学到的，将为我做好准备目标 3 中提出的实验将探索其他 CSF 传播的作用和机制信号因子，如视黄酸和 Sonic Hedgehog，受 CSF 调节作用于靶细胞到本奖励期结束时，我的独立研究实验室将拥有开创了发育生物学的新概念，其中脑脊液在大脑皮层中发挥积极作用所提出的实验为未来研究变化奠定了坚实的基础。由于脑脊液是一种外科手术，因此脑脊液干细胞在衰老和与年龄相关的神经系统疾病中发挥着重要作用。在人类和小鼠模型中可访问的介质中，所提出的实验将为开发强大的诊断和治疗方法。