Intercellular TWEAK/Fn14 Cytokine Signaling in Sensory-Dependent Circuit Refinement

感觉依赖性电路细化中的细胞间 TWEAK/Fn14 细胞因子信号转导

基本信息

批准号：
9754316
负责人：
Lucas M Cheadle
金额：
$ 10.5万
依托单位：
HARVARD MEDICAL SCHOOL
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-03-14 至 2021-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9754316
关键词：
Acute Address Binding Bioinformatics Biological Process Brain CRISPR/Cas technology Cell Nucleus Cells Complement Complex Cues Cytokine Receptors Cytokine Signaling Data Dendritic Spines Development Disease Electrophysiology (science)Environment Environmental Risk Factor Foundations Functional disorder Gene Expression Genes Genetic Genetic Transcription Goals Image Immersion Investigative Technique Immune Impairment Inflammatory Institution Intellectual functioning disability Interdisciplinary Study Knowledge Lead Learning Life Ligands Mediating Mentorship Methods Microglia Modeling Molecular Mus Nature Neurodevelopmental Disorder Neurons Neurophysiology - biologic function Phagocytes Phase Photic Stimulation Positioning Attribute Process Research Resolution Role Scientist Sensory Signal Pathway Signal Transduction Slice Structure Synapses Techniques Testing Therapeutic Training Training Activity Vertebral column Viral Visual Work career cell type course development critical period cytokine excitatory neuron experience experimental study gain of function gene induction human disease immune activation in vivo innovation insight loss of function medical schools mouse model neural circuit neurodevelopment novel postnatal postnatal development postnatal period programs psychiatric disability relating to nervous system response sensory input single cell sequencing single-cell RNA sequencing skills transcriptomics two-photon

项目摘要

During postnatal brain development, newly assembled neural circuits are refined through the strengthening of a subset of synaptic connections and the concurrent elimination of others. This process of synaptic refinement is first coordinated by intrinsically generated neural activity early in life and then driven by sensory experience during a later phase of postnatal development. Impairments in sensory-dependent refinement are thought to contribute to a heterogeneous array of neurodevelopmental disorders, consistent with the contribution of environmental risk factors, such as immune challenge, to their pathophysiology. Yet, therapeutic strategies aimed at correcting such impairments have been limited by a fundamental lack of insight into the molecular and cellular mechanisms through which sensory input refines developing circuits. The key focus of the proposed research is to obtain a more comprehensive understanding of the basic cellular and molecular mechanisms by which sensory experience refines synaptic connections in the course of development, and to determine how impairments in these mechanisms may contribute to neurodevelopmental disease. In preliminary studies, the applicant applied newly developed single-cell transcriptomic methods to identify Fn14 as a gene that is transcriptionally induced by visual experience in excitatory neurons and encodes a cytokine receptor that is required for sensory-dependent refinement in the developing brain. This proposal contains a comprehensive training and research plan to build upon these findings by addressing the remaining gaps in knowledge experimentally in the short-term, and by facilitating the establishment of an innovative and multidisciplinary research program focused on addressing these questions in the long-term. In Aim 1, the applicant will learn and apply specialized acute slice electrophysiology techniques to test the hypothesis that TWEAK, the pro-inflammatory cytokine ligand of Fn14, is necessary and sufficient to drive sensory-dependent refinement via a microglia-to-neuron signaling axis. In Aim 2, the applicant will combine electrophysiology with newly learned methods in bioinformatics to determine whether heightened TWEAK/Fn14 signaling contributes to excessive synaptic refinement in the maternal immune activation (MIA) mouse model of aberrant brain development. In Aim 3, the applicant will test the hypothesis that TWEAK/Fn14 signaling mediates functional refinement by promoting the conversion of immature synaptic spines to mature spines. These technical training activities will be augmented by focused mentorship from several highly successful scientists who are committed to aiding in the applicant's acquisition of professional and intellectual skills in the highly energetic and collaborative training environment at Harvard Medical School. These activities are expected to facilitate the successful transition of the applicant into an independent research position at an academic institution.

在出生后大脑发育过程中，新组装的神经回路通过加强一部分突触连接并同时消除其他连接而得到完善。这种突触细化的过程首先由生命早期内在产生的神经活动协调，然后在出生后发育的后期阶段由感觉体验驱动。感觉依赖性细化的损伤被认为会导致一系列异质性神经发育障碍，这与免疫挑战等环境风险因素对其病理生理学的影响一致。然而，旨在纠正此类损伤的治疗策略由于对感觉输入改善发育回路的分子和细胞机制缺乏深入了解而受到限制。拟议研究的重点是更全面地了解感官体验在发育过程中细化突触连接的基本细胞和分子机制，并确定这些机制的损伤如何导致神经发育疾病。在初步研究中，申请人应用新开发的单细胞转录组学方法将Fn14鉴定为兴奋性神经元中视觉体验转录诱导的基因，并编码发育中大脑中感觉依赖性细化所需的细胞因子受体。该提案包含一个全面的培训和研究计划，以这些发现为基础，通过短期实验解决知识方面的剩余差距，并促进建立一个创新的多学科研究计划，重点关注长期解决这些问题。在目标 1 中，申请人将学习并应用专门的急性切片电生理学技术来测试以下假设：TWEAK（Fn14 的促炎细胞因子配体）对于通过小胶质细胞到神经元信号传导轴驱动感觉依赖性细化是必要且充分的。在目标 2 中，申请人将把电生理学与生物信息学中新学到的方法结合起来，以确定增强的 TWEAK/Fn14 信号传导是否有助于异常大脑发育的母体免疫激活 (MIA) 小鼠模型中突触的过度细化。在目标 3 中，申请人将测试 TWEAK/Fn14 信号传导通过促进未成熟突触棘向成熟突触棘的转化来介导功能细化的假设。这些技术培训活动将得到几位非常成功的科学家的集中指导，他们致力于帮助申请人在哈佛医学院充满活力和协作的培训环境中获得专业和智力技能。这些活动预计将促进申请人成功过渡到学术机构的独立研究职位。