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来自我在宾夕法尼亚大学的本科教育，在那里，我是Art McMorris博士的Wistar Institute本科生，我研究了GLIA中的外部提示触发营地信号。作为Bruce Mcewen博士实验室洛克菲勒大学的本科研究员，我研究了甲状腺功能减退症如何改变大脑的信号分子。这些经历使我曾在哈佛大学学习研究生院，在那里我曾在分子神经科学和阐明信号通路方面与阿扎德·邦尼（Azad Bonni）博士进行了培训，这些信号通路介导了哺乳动物神经元和秀丽隐杆线虫中体内的生存和死亡信号传导。博士学位完成后，我在芬兰赫尔辛基的Folkhdlsan遗传学研究所与Anna-Elina Lehesjoki博士进行了早期的博士后培训，在那里我发现氧化还原稳态受损是一种关键的机制，是一种促进肌lon症epilepsy的神经变性的关键机制。从这些研究经验中，我在分子神经科学及其在神经系统疾病中的应用中获得了经验。我与克里斯·沃尔什（Chris Walsh）博士的实验室一起参加了我的第二个博士后，希望在该奖项的指导阶段进行发育神经生物学方面的全面培训将补充我在分子神经科学方面的培训，并就整个生命的神经学疾病的基本机制提供独特的观点。 沃尔什（Walsh's）博士是脑皮质发展研究领域的领导者。我将学习实验室中的基本技术，从定量组织学分析到子宫电气。沃尔什（Walsh）博士召集了一群才华横溢的科学家团体，该小组将继续在非正式的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（Walsh）实验室位于遗传学部门，该部门还与神经科学部进行了每周共同的数据对话，以促进协作交流和有关技术和理论问题的协助。在奖项的指导阶段，这种吸引人的环境将提供出色的讨论和反馈来源。重要的是，遗传学的划分致力于支持博士后向独立研究职位的过渡。为此，我将参加许多职业培训研讨会和会议，旨在为我启动独立研究职业做好准备。 我的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，该信号传导位置可以协调广泛的信号因子相互作用，并在长途行动以调节健康和疾病期间的靶细胞行为。在脑皮质发育过程中，在神经上皮的广大地区，增殖细胞的快速变化几乎同步发生。神经上皮纤毛在CSF中的长期位置表明，CSF作为指导祖细胞增殖的外部信号来源的潜在作用。由于CSF每天翻转几次，因此CSF-螺旋 - 棕榈系统非常适合触发跨大距离的分子信号的快速和空间同步变化。我在该提案中的直接研究目标是研究胚胎CSF蛋白质组在调节发育过程中皮质祖细胞增殖中的作用。 AIM1中的实验将使用异缘植体和培养的干细胞来测试CSF支持皮质祖细胞生存，生长和增殖的能力。由于CSF含有数百种蛋白质，因此我将使用生化和遗传方法来检查IGF2（胰岛素样生长因子2）如何在我们的初步质谱分析中鉴定出的候选因子，可以通过CSF主动分布CSF来影响祖先（AIM 2）。该奖项的指导阶段（AIMS 1和2）将有助于完善成功，独立的研究计划所需的实验模型，技术和专业技能，研究CSF分布的因素影响健康和疾病中目标细胞的机制。 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.到这个奖励期结束时，我的独立研究实验室将开创一个新的概念，其中CSF在脑皮质发育中发挥了积极作用。提出的实验代表了未来研究的稳固基础，研究了衰老和与年龄相关的神经系统疾病中CSF干细胞生态裂的变化。由于CSF是人类和小鼠模型中的外科手术介质，因此提出的实验将为开发强大的诊断和治疗方法开发铺平道路。