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.

项目总结/摘要 我对神经系统疾病的兴趣源于我在芝加哥大学的本科教育。 在宾夕法尼亚州，作为一个威斯塔研究所的本科生研究员与博士艺术麦克莫里斯，我调查了外在的 神经胶质细胞中触发cAMP信号的线索。作为洛克菲勒大学的一名本科研究员， 在布鲁斯·麦克尤恩的实验室，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 这些经历使我进入了哈佛大学的研究生院，在那里我与阿扎德·邦尼博士一起学习分子生物学。 神经科学和阐明的信号通路，介导的生存和死亡信号在哺乳动物 神经元和体内C.优雅的完成博士学位后，我进行了早期的博士后培训 我和芬兰赫尔辛基Folkh úlsan遗传学研究所的Anna-Elina Lehesjoki博士在一起， 受损的氧化还原稳态是引发进行性神经退行性变的关键机制， 肌阵挛性癫痫，Unverricht-Lundborg病（EPM 1）。从这些研究经验中，我获得了 在分子神经科学及其在神经疾病中的应用方面的经验。我加入了克里斯沃尔什博士的实验室 我的第二个博士后，希望在发展神经生物学的全面培训期间， 这个奖项的指导阶段将补充我在分子神经科学的培训，并提供独特的 在整个生命的神经系统疾病的潜在机制的观点。 沃尔什博士是大脑皮层发育研究领域的领导者。我将学习基础 实验室中的技术，从定量组织学分析到子宫内电穿孔。沃尔什医生 汇集了一批才华横溢的科学家，他们将继续提供独特的支持来源， 在非正式的谈话和每周的实验室会议和杂志俱乐部的灵感。沃尔什实验室位于 遗传学部还与神经科学部联合举办每周数据会谈，以促进 合作交流和技术和理论知识援助。这种智力上的吸引力 环境将提供一个很好的讨论和反馈的来源，在指导阶段， 奖重要的是，遗传学系致力于支持博士后过渡到 独立研究岗位。为此，我将参加一些职业培训研讨会和会议 旨在为我开始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， 一个信号小生境，协调信号因子的丰富相互作用，在长距离作用，调节靶向 健康和疾病期间的细胞行为。在大脑皮层发育过程中， 祖细胞几乎同步地出现在神经上皮的大面积上。延长的位置 CSF中神经上皮纤毛的存在表明CSF作为外源性信号引导的潜在作用。 祖细胞增殖。由于CSF每天翻转数次，因此CSF-脉络丛系统 理想地适合于触发大分子信号传导中的快速和空间同步的变化， 的距离.我在该提案中的直接研究目标是研究胚胎CSF蛋白质组在以下方面的作用： 在发育过程中调节皮质祖细胞增殖。 Aim 1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。 以支持皮质祖细胞的存活、生长和增殖。由于脑脊液中含有数百个 然后，我将使用生物化学和遗传学方法来研究Igf 2（胰岛素样生长因子） 2），在我们的初步质谱分析中鉴定的候选因子，可以通过 CSF以影响祖细胞增殖（Aim 2）。该奖项的指导阶段（目标1和2）将有助于 完善实验模型，技术和专业技能，需要推出一个成功的， 一项独立的研究计划，研究CSF分布因素影响目标的机制 健康和疾病中的细胞。基本技术包括神经球培养、定量 组织学分析，子宫内电穿孔，所有这些都是在目标1&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携带的 信号传导因子，如视黄酸和Sonic hedgehog，由CSF调节作用于靶细胞， 心室表面。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CSF在大脑皮层中起着积极的作用。 发展所提出的实验代表了一个坚实的基础，为未来的研究调查的变化 在衰老和与年龄相关的神经系统疾病中的CSF干细胞生态位中。由于脑脊液是一种外科手术 在人类和小鼠模型中，拟议的实验将为 开发强大的诊断和治疗方法。