Probing Microtubule Function in Neuronal Development

探索神经元发育中的微管功能

基本信息

批准号：
10116503
负责人：
Torsten Wittmann
金额：
$ 35.84万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-06-15 至 2023-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10116503
关键词：
Address Binding Brain CRISPR/Cas technology Cells Cellular biology Clinical Complex Cortical Malformation Cytoskeleton Data Date of birth Destinations Development Disease Filament Foundations Future Genes Genome engineering Geometry Growth Cones Health Human Human Genetics Impairment Link Maps Mechanics Mediating Microtubule-Associated Proteins Microtubules Missense Mutation Modeling Modernization Molecular Morphogenesis Morphology Mutate Mutation Names Nervous System Physiology Nervous system structure Neurites Neurobiology Neurodegenerative Disorders Neuronal Differentiation Neurons Organism Outcome Phenotype Physiological Plus End of the Microtubule Proteins Quantitative Microscopy Research Specificity Structure System Techniques Technology Tertiary Protein Structure Testing Therapeutic Tubulin base brain malformation cell type contrast enhanced excitatory neuron genome editing human pluripotent stem cell induced pluripotent stem cell induced pluripotent stem cell technology information processing innovation insight lissencephaly migration nervous system development nervous system disorder neurogenesis neuron development newborn neuron novel optogenetics protein complex recruit synaptogenesis tool

项目摘要

Neurons are the most morphologically complex cell type in multicellular organisms, and this complexity is intricately linked to the complexity of nervous system information processing. Thus, impairments in neuronal morphogenesis invariably result in alterations of nervous system function and often debilitating neurological disease. Precise control of microtubule cytoskeleton organization is central to most if not all aspects of neuron development and function ranging from the migration of newborn neurons, cycles of neurite elongation, retraction and branching to growth cone guidance and synapse formation. This importance of neuronal microtubules is reflected in the wide range of neurodevelopmental and neurodegenerative diseases linked to genetic defects in proteins associated with the microtubule cytoskeleton. Human genetics and modern sequencing techniques identified numerous mutations in related genes associated with frequently severe cortical malformations. These include both mutations of tubulin genes themselves – so-named tubulinopathies – as well as mutations in neuronal microtubule-associated proteins that often have a similar range of neurodevelopmental phenotypes. While it is generally assumed that these mutations disrupt the developmental migration of immature neurons through the developing cortex, we do not understand the rules governing MT function in neuromorphogenesis at a mechanistic level. In this application, we propose to use emerging human induced pluripotent stem cell (iPSC) technology in combination with state-of-the-art genome engineering and our established expertise in quantitative microscopy to model and dissect the neuronal cell biology of tubulinopathy-like diseases and the function of dynamic MTs in neuromorphogenesis. In Aim 1, we focus on DCX and tubulin mutations that cause cortical malformations, and we will analyze how DCX controls neuronal microtubule dynamics and mechanics, and neuronal morphogenesis based on our recent data that DCX binds microtubules in a unique geometry-dependent way. In Aim 2, we employ novel optogenetics to control protein interactions with growing microtubule plus ends with second and micrometer precision to map how microtubule plus end complexes contribute to neuronal development dynamics. We believe that quantitative and rigorous understanding of the principles that govern MT function in neuronal development and what goes wrong in neurodevelopmental disease will have tangible and important outcomes for human health, and can lay the foundation for future therapeutic approaches.

神经元是多细胞生物中形态上最复杂的细胞类型，这种复杂性是与神经系统信息处理的复杂性相关。那是神经元的损害形态发生总是会导致神经系统功能的改变，并经常使神经系统衰弱疾病。微管细胞骨架组织的精确控制是神经元的大多数（即使不是全部）的核心发展和功能范围从新生神经元的迁移，神经元的周期，缩回和分支以增加锥形指导和突触形成。神经元的重要性微管反映在与与之相关的广泛神经发育和神经退行性疾病中与微管细胞骨架相关的蛋白质中的遗传缺陷。人类遗传学和现代测序技术确定了与经常严重相关的相关基因中的许多突变皮质畸形。这些包括小管蛋白基因本身的突变 - 所谓的小管状病毒 - 以及神经元微管相关蛋白的突变，这些蛋白通常具有相似的范围神经发育表型。虽然通常认为这些突变破坏了发展未成熟神经元通过发育中的皮质迁移，我们不了解管理的规则 MT在机械水平上神经形成中的功能。在此应用程序中，我们建议使用新兴人类诱导的多能干细胞（IPSC）技术与最先进的基因组结合工程和我们在定量显微镜方面已建立的专业知识，以建模和剖析神经元细胞微管疾病样疾病的生物学以及动态MT在神经形态发生中的功能。在AIM 1中，我们专注于引起皮质畸形的DCX和微管蛋白突变，我们将分析DCX如何控制神经元微管动力学和力学，以及基于我们最近的数据，神经元形态发生 DCX以独特的几何学方式结合微管。在AIM 2中，我们采用新颖的光遗传学控制蛋白质与生长的微管加的相互作用，并以第二和千分尺的精度结束微管加复合物如何有助于神经元发展动力学。我们相信对控制MT在神经元发展中功能的原理的定量和严格理解神经发育疾病中出了什么问题，将对人类健康产生切实而重要的结果，并可以为未来的治疗方法奠定基础。