The Role of Fos and the BAF Complex in Neuronal Activity-Dependent Chromatin Remodeling and Gene Expression

Fos 和 BAF 复合物在神经元活动依赖性染色质重塑和基因表达中的作用

• 批准号: 10572785
• 负责人: Sara Trowbridge
• 金额: $ 19.4万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10572785
• 关键词: Advisory Committees; Affect; Animal Model; Area; Binding; Binding Sites; Brain; CRISPR/Cas technology; Cells; Childhood; Chromatin; Chromatin Remodeling Factor; Collection; Complex; Data; Development; Developmental Process; Disease; Disease model; Distal; Distal Enhancer Elements; Enhancers; Epilepsy; Ethics; Exhibits; Fibroblasts; Functional disorder; Gene Expression; Genes; Genetic; Genetic Databases; Genetic Transcription; Genetic Variation; Genomics; Heterozygote; Human; Human Activities; Human Genetics; Human Genome; In Vitro; Individual; Intellectual functioning disability; Knowledge; Laboratories; Life; Light; Mediating; Mentors; Mentorship; Modeling; Molecular; Morbidity - disease rate; Mus; Neurodevelopmental Disorder; Neuronal Plasticity; Neurons; Neurosciences; Neurosciences Research; Nucleic Acid Regulatory Sequences; Pathogenesis; Pathway interactions; Patients; Pediatric Neurology; Phenotype; Physicians; Physiological; Play; Positioning Attribute; Process; Program Development; Regulation; Regulator Genes; Regulatory Element; Regulon; Research; Role; Scientist; Site; Source; Stimulus; Techniques; Technology; Testing; Therapeutic; Time; Training; Variant; Visual Cortex; Work; autism spectrum disorder; behavioral phenotyping; brain abnormalities; career; career development; cell type; childhood epilepsy; chromatin remodeling; dbSNP; design; developmental disease; experimental study; genetic variant; human disease; human embryonic stem cell; human embryonic stem cell line; human model; in vitro Model; in vivo; in vivo Model; insight; mouse model; neural circuit; neurodevelopment; neurogenetics; new therapeutic target; next generation sequencing; novel therapeutics; programs; recruit; response; single-cell RNA sequencing; synaptic pruning; transcription factor; translational neuroscience

Pediatric neurodevelopmental disorders (NDD), including pediatric epilepsy, autism spectrum disorder, and intellectual disability, represent a major source of morbidity, yet our therapeutic options remain limited. Development of novel therapies for NDD will require a deeper mechanistic understanding of normal and abnormal brain development. This proposal focuses on genetic programs that are activated in response to neuronal activity and that are fundamental to normal neurodevelopment and on-going neuronal plasticity throughout life. Fos is a major activity-dependent transcription factor that binds to distal enhancer elements and regulates downstream activity-dependent genetic programs in a cell-type-specific manner to promote key processes, including synaptic pruning and the recruitment of inhibition in the developing brain. Despite its role in key developmental processes, we do not understand how Fos is differentially targeted to cell-type-specific binding sites, nor how genetic variation at these sites impacts neurodevelopment and neuronal function. Interestingly, Fos has been shown to physically interact with the BAF chromatin remodeling complex, and it is possible that this interaction is critical to Fos function. Many BAF subunits, most frequently ARID1B, are implicated in human NDD, but whether the BAF complex regulates neuronal activity-dependent genetic programs, and how this underlies aspects of BAF complex-related NDD, is previously unexplored. The research in this proposal will address these gaps in knowledge by: (a) profiling Fos and BAF complex neuronal binding sites across the human genome; (b) assessing human genetic variants at these sites in individuals with NDD vs controls; and (c) determining the effects of BAF complex perturbation on neuronal activity-dependent genetic programs in vitro and in vivo. Overall, this work will lead to greater insight into how activity-dependent genetic programs contribute to NDD pathogenesis. Additionally, by identifying specific activity-regulated genes and pathways that are mis-regulated downstream of Fos and the BAF complex, these experiments could highlight novel therapeutic targets for NDD. This research is the basis for a five-year career development program designed to build on Dr. Trowbridge’s background in molecular neuroscience, pediatric neurology/epilepsy, and neurogenetics, by providing her with additional training in analysis of human sequencing data, use of in vitro and in vivo models of NDD, and next- generation sequencing technologies. Her primary mentor, Dr. Mike Greenberg, and her scientific advisory committee, Drs. Annapurna Poduri and Chris Walsh, will provide guidance in these areas, as well as mentorship in the rigorous and ethical conduct of translational neuroscience research. Thus the proposed training plan will position Dr. Trowbridge to launch her independent career as a clinician-scientist focused on understanding the role of activity-dependent genetic programs in NDD.

小儿神经发育障碍 (NDD)，包括小儿癫痫、自闭症谱系障碍和 智力障碍是发病的一个主要来源，但我们的治疗选择仍然有限。 NDD 的新疗法的开发需要对正常和 大脑发育异常。该提案的重点是响应以下因素而激活的基因程序： 神经元活动，是正常神经发育和持续神经元可塑性的基础 一生。 Fos 是一种主要的活性依赖性转录因子，可与远端增强子元件结合并 以细胞类型特异性方式调节下游活动依赖性遗传程序，以促进关键 过程，包括突触修剪和发育中大脑中抑制的招募。尽管它的作用 关键的发育过程，我们不明白 Fos 如何针对细胞类型特异性进行差异化靶向 结合位点，以及这些位点的遗传变异如何影响神经发育和神经元功能。 有趣的是，Fos 已被证明与 BAF 染色质重塑复合物发生物理相互作用，并且 这种相互作用可能对 Fos 功能至关重要。许多 BAF 亚基，最常见的是 ARID1B， 与人类NDD有关，但BAF复合物是否调节神经元活动依赖的遗传 程序，以及这如何成为 BAF 复合体相关 NDD 的基础，此前尚未被探索过。研究 该提案将通过以下方式解决这些知识空白：(a) 分析 Fos 和 BAF 复杂的神经元结合 人类基因组中的位点； (b) 评估 NDD 个体与 NDD 个体在这些位点的人类遗传变异 控制； (c) 确定 BAF 复合体扰动对神经元活动依赖性遗传的影响 体外和体内程序。总体而言，这项工作将导致人们更深入地了解活动依赖性遗传如何 计划有助于 NDD 发病机制。此外，通过识别特定的活性调节基因和 Fos 和 BAF 复合体下游被错误调节的通路，这些实验可以突出显示 NDD 的新治疗靶点。 这项研究是五年职业发展计划的基础，该计划旨在以特罗布里奇博士的 分子神经科学、儿科神经病学/癫痫和神经遗传学背景，为她提供 人类测序数据分析、NDD 体外和体内模型的使用以及下一步的额外培训 世代测序技术。她的主要导师迈克·格林伯格博士和她的科学建议 委员会，博士。 Annapurna Poduri 和 Chris Walsh 将在这些领域提供指导和辅导 转化神经科学研究的严格和道德行为。因此，拟议的培训计划将 特罗布里奇博士作为一名临床医生科学家开启了她的独立职业生涯，专注于了解 活动依赖性遗传程序在 NDD 中的作用。