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: 8129458
• 负责人: MARIA LEHTINEN
• 金额: $ 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 2012-01-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8129458
• 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; Education; 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; Nerve Degeneration; Neuroepithelial; Neuroglia; Neurologic; Neurons; Neurosciences; Oxidation-Reduction; Pattern; Pennsylvania; Phase; Play; Positioning Attribute; Postdoctoral Fellow; Process; Progressive Myoclonic Epilepsies; 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; 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

DESCRIPTION (provided by applicant): 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 Folkhdlsan Institute of Genetics in Helsinki, Finland, where I discovered that impaired redox homeostasis is a key mechanism triggering neurodegeneration in the progressive 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 issues. 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, will 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 will 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.

描述（由申请人提供）： 我对神经系统疾病的兴趣源于我在宾夕法尼亚大学的本科教育，在那里，作为Wistar研究所的本科研究员，我与Art McMorris博士一起研究了触发神经胶质细胞中cAMP信号的外在线索。作为洛克菲勒大学布鲁斯·麦克尤恩博士实验室的一名本科研究员，我研究了甲状腺功能减退症如何改变大脑中的信号分子。这些经历使我进入了哈佛大学的研究生院，在那里我与Azad Bonni博士一起接受了分子神经科学的培训，并阐明了哺乳动物神经元和体内C.优美的完成博士学位后，我在芬兰赫尔辛基的Folkhdlsan遗传学研究所与Anna-Elina Lehesjoki博士进行了早期博士后培训，在那里我发现氧化还原稳态受损是引发进行性肌阵挛癫痫，Unverricht-Lundborg病（EPM 1）神经变性的关键机制。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应用的经验。我加入了克里斯沃尔什博士的实验室，我的第二个博士后，希望在这个奖项的指导阶段发育神经生物学的全面培训将补充我在分子神经科学的培训，并提供对整个生命的神经疾病的机制的独特视角。 沃尔什博士是大脑皮层发育研究领域的领导者。我将在实验室学习基本技术，从定量组织学分析到子宫内电穿孔。沃尔什博士召集了一批才华横溢的科学家，他们将继续在非正式对话和每周实验室会议和期刊俱乐部中提供独特的支持和灵感来源。沃尔什实验室位于遗传学部，该部门还与神经科学部每周举行联合数据会谈，以促进技术和理论问题的合作交流和援助。这种智力参与的环境将提供一个很好的讨论和反馈的来源，在指导阶段的奖励。重要的是，遗传学系致力于支持博士后向独立研究职位的过渡。为此，我将参加一些职业培训研讨会和会议，为我的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）如何作为一个信号微环境发挥作用，协调信号因子的丰富相互作用，在健康和疾病期间长距离发挥作用以调节靶细胞行为。在大脑皮质发育过程中，增殖祖细胞的快速变化几乎同步发生在神经上皮的大面积。神经上皮纤毛在CSF中的延长位置表明CSF作为引导祖细胞增殖的外源信号的来源的潜在作用。由于CSF每天翻转几次，CSF-脉络丛系统非常适合于触发跨越大距离的分子信号传导的快速和空间同步变化。我的近期研究目标是研究胚胎CSF蛋白质组在发育过程中调节皮质祖细胞增殖的作用。 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支持皮质祖细胞存活、生长和增殖的能力。由于CSF含有数百种蛋白质，因此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确定的候选因子）如何通过CSF积极分布以影响祖细胞增殖（目的2）。该奖项的指导阶段（目标1&2）将有助于完善实验模型，技术和专业技能，以启动一个成功的，独立的研究计划，研究CSF分布因子影响健康和疾病靶细胞的机制。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中学习的，将为目标3中提出的实验做好准备，该实验将探索其他CSF携带的信号传导因子（如视黄酸和Sonic hedgehog）受CSF调节作用于心室表面靶细胞的作用和机制。在这个奖励期结束时，我的独立研究实验室将在发育生物学中开创一个新的概念，即CSF在大脑皮层发育中起着积极的作用。拟议的实验代表了一个坚实的基础，为未来的研究调查在老化和年龄相关的神经系统疾病的CSF干细胞生态位的变化。由于CSF是人类和小鼠模型中可通过手术获得的介质，因此拟议的实验将为开发强大的诊断和治疗方法铺平道路。