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.

儿童神经发育障碍（NDD），包括儿童癫痫、自闭症谱系障碍和 智力残疾是发病主要原因，但我们的治疗选择仍然有限。 NDD的新疗法的开发将需要对正常和 大脑发育异常这项建议的重点是基因程序，激活响应 神经元活动，是正常神经发育和持续神经元可塑性的基础 在生活中。Fos是一种主要的活性依赖性转录因子，它与远端增强子元件结合， 以细胞类型特异性方式调节下游活性依赖性遗传程序， 过程，包括突触修剪和发育中的大脑抑制的招募。尽管它在 关键的发育过程，我们不知道如何Fos是差异靶向细胞类型特异性 结合位点，也不知道这些位点的遗传变异如何影响神经发育和神经功能。 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， 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ARID 1B， 与人类NDD有关，但BAF复合物是否调节神经元活性依赖性遗传 程序，以及如何这方面的BAF复杂相关的NDD的基础，是以前未探索。研究 在这项提案中，将通过以下方式解决这些知识空白：（a）分析Fos和BAF复合物神经元结合 （B）评估患有NDD的个体中这些位点处的人类遗传变异， （c）确定BAF复合物扰动对神经元活性依赖性遗传的影响， 在体外和体内的程序。总的来说，这项工作将使我们更深入地了解活动依赖性遗传 程序有助于NDD发病机制。此外，通过鉴定特定的活性调节基因， 这些实验可以强调Fos和BAF复合物下游错误调节的途径 NDD的新治疗靶点。 这项研究是一个为期五年的职业发展计划的基础，旨在建立在特罗布里奇博士的 分子神经科学，儿科神经病学/癫痫和神经遗传学的背景，通过为她提供 在分析人类测序数据、使用体外和体内NDD模型方面进行额外培训，接下来- 代测序技术。她的主要导师迈克·格林伯格博士和她的科学顾问 委员会，安娜普纳波杜里博士和克里斯沃尔什，将提供指导，在这些领域，以及导师 在严格和道德的行为转化神经科学研究。因此，拟议的培训计划将 特罗布里奇博士的立场，开始她作为一个临床科学家的独立职业生涯，专注于了解 活动依赖性遗传程序在NDD中的作用。