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

• 批准号: 8402006
• 负责人: MARIA LEHTINEN
• 金额: $ 24.9万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 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/8402006
• 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

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.

项目摘要/摘要 我对神经系统疾病的兴趣源于我在大学的大学教育 宾夕法尼亚州，在Art McMorris博士的Wistar Institute Begrance中，我调查了外部 提示在神经胶质中触发营地信号。作为博士洛克菲勒大学的本科研究员 布鲁斯·麦克尤恩（Bruce McEwen）的实验室，我研究了甲状腺功能减退症如何改变大脑中的信号分子。这些 经验使我在哈佛大学学习了研究生院，在那里我曾在分子的Azad Bonni博士培训过 神经科学和阐明培养基生存和死亡信号传导的信号通路 神经元和秀丽隐杆线虫的体内。博士学位完成后，我进行了早期的博士后培训 与Anna-Elina Lehesjoki博士一起，在芬兰赫尔辛基的Folkh。LSAN遗传学研究所，在那里我发现了 氧化还原稳态受损是触发神经变性的关键机制 Myoclonus癫痫，Unverricht-Lundborg病（EPM1）。从这些研究经验中，我获得了 分子神经科学及其在神经系统疾病中的应用。我加入了克里斯·沃尔什博士的实验室 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培训 该奖项的指导阶段将完成我在分子神经科学方面的培训，并提供独特 一生中神经系统疾病的机制的观点。 沃尔什（Walsh's）博士是脑皮质发展研究领域的领导者。我将学习基本 实验室中的技术，从定量组织学分析到子宫电气。沃尔什博士有 组建了一群才华横溢的科学家群体，该群体将继续提供独特的支持来源和 非正式对话和每周实验室会议和期刊俱乐部中的灵感。沃尔什实验室位于 遗传学的划分（还与神经科学分工进行了联合的每周数据对话），以促进 在技​​术和理论问题上进行协作交流和协助。这很聪明 环境将在修订阶段的讨论和反馈中提供极好的来源 奖。重要的是，遗传学的划分致力于支持博士后过渡到 独立的研究职位。为此，我将参加许多职业培训半手和会议 旨在为我启动独立研究生涯做好准备。 我的研究的长期目标是阐明脑脊液（CSF）如何用作 信号的利基市场可以协调广泛的信号因子相互作用，以长距离作用以调节目标 健康和疾病期间的细胞行为。在脑皮质发育期间，增殖的快速变化 祖细胞几乎同步地发生在神经上皮的广大区域。旷日持久的位置 CSF中的神经上皮纤毛表明，CSF作为外在信号的来源的潜在作用 祖细胞增殖。由于CSF每天翻过来几次 非常适合触发大型分子信号的快速和空间同步变化 距离。我在此提案中的直接研究目标是调查胚胎CSF蛋白质组在 在发育过程中调节皮质祖细胞增殖。 AIM1中的实验将使用异缘植体和培养的干细胞测试CSF的能力 支持皮质祖细胞的生存，生长和增殖。由于CSF包含数百个 然后，我将使用生化和遗传方法来检查IGF2（胰岛素样生长因子） 2）在我们的初步质谱分析中确定的候选因素，可以通过 CSF影响祖细胞增殖（AIM 2）。奖项的重要阶段（目标1和2）将有助于 完善推出成功的实验模型，技术和专业技能 独立研究计划，研究了CSF分布因素影响目标的机制 健康和疾病中的细胞。基本技术在内，包括神经球培养，定量 组织学分析和子宫电气中，所有在目标1和2中都学会了，将为我做好准备 AIM 3中提出的实验将探讨其他CSF裔裔的作用和机制 信号因子（例如视黄酸和声音刺猬）受到CSF的调节，以作用于目标细胞 心室表面。到这个颁奖期结束时，我的独立研究实验室将 开创了一个新的概念，在发育生物学中，脑脊液在脑皮质中发挥了积极作用 发展。拟议的实验代表了未来研究的坚实基础，研究了变化 在衰老和与年龄相关的神经系统疾病中的CSF干细胞生态位。由于CSF是外科手术 在人类和鼠标模型中可访问的介质，提出的实验将为迈向 开发强大的诊断和治疗方法。