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: 8028164
• 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 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8028164
• 关键词: Adult; Affect; Age; 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; 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; progenitor; programs; public health relevance; 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接受的本科教育，作为 Wistar 研究所的本科生，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生研究员，我研究了甲状腺功能减退症如何改变大脑中的信号分子。这些经历使我进入哈佛大学研究生院，在那里我与 Azad Bonni 博士一起接受分子神经科学方面的培训，并阐明了介导哺乳动物神经元和线虫体内生存和死亡信号传导的信号通路。完成博士学位后，我在芬兰赫尔辛基 Folkhdlsan 遗传学研究所与 Anna-Elina Lehesjoki 博士一起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中神经退行性变的关键机制， Unverricht-Lundborg 病 (EPM1)。从这些研究经历中，我获得了分子神经科学及其在神经系统疾病中的应用的经验。我加入 Chris Walsh 博士的实验室进行我的第二个博士后工作，希望在该奖项的指导阶段接受的发育神经生物学综合培训能够补充我在分子神经科学方面的培训，并为整个生命过程中神经系统疾病的机制提供独特的视角。 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。沃尔什博士组建了一支才华横溢的科学家团队，他们将继续在非正式对话、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门，该部门还与神经科学部门每周举办联合数据会谈，以促进技术和理论问题上的合作交流和协助。这种充满智力的环境将为奖项的指导阶段提供一个极好的讨论和反馈来源。重要的是，遗传学部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）如何作为信号传导生态位发挥作用，协调信号传导因子的丰富相互作用，在健康和疾病期间远距离发挥作用来调节靶细胞行为。在大脑皮层发育过程中，增殖祖细胞的快速变化几乎同时发生在神经上皮的广大区域。神经上皮纤毛在脑脊液中的长期定位表明脑脊液作为引导祖细胞增殖的外在信号源的潜在作用。由于脑脊液每天翻转数次，因此脑脊液-脉络丛系统非常适合触发远距离分子信号传导的快速且空间同步的变化。我在本提案中的直接研究目标是研究胚胎脑脊液蛋白质组在发育过程中调节皮质祖细胞增殖的作用。 Aim1 中的实验将使用异时外植体和培养的干细胞来测试脑脊液支持皮质祖细胞存活、生长和增殖的能力。由于脑脊液含有数百种蛋白质，因此我将使用生化和遗传学方法来检查 Igf2（胰岛素样生长因子 2）（我们初步质谱分析中确定的候选因子）如何由脑脊液主动分布以影响祖细胞扩散（目标 2）。该奖项的指导阶段（目标 1 和 2）将有助于完善启动成功的独立研究项目所需的实验模型、技术和专业技能，以检查脑脊液分布的因子影响健康和疾病中靶细胞的机制。包括神经球培养、定量组织学分析和子宫内电穿孔在内的基本技术，所有这些都是在目标 1 和 2 期间学到的，将为我在目标 3 中提出的实验做好准备，该实验将探索其他 CSF 传播的信号因子的作用和机制，例如如视黄酸和音刺猬，由脑脊液调节作用于心室表面的靶细胞。到本奖励期结束时，我的独立研究实验室将开创发育生物学的新概念，其中脑脊液在大脑皮层发育中发挥积极作用。拟议的实验为未来研究脑脊液干细胞生态位在衰老和年龄相关神经系统疾病中的变化奠定了坚实的基础。由于脑脊液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。