Examining Microglial Environmental Dependent Transcriptional Networks

检查小胶质细胞环境依赖性转录网络

• 批准号: 10241987
• 负责人: Nicole Gabriele Coufal
• 金额: $ 20.5万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-30 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10241987
• 关键词: ATAC-seq; Aging; Astrocytes; Atlases; Award; Biological Assay; Brain; CCL2 gene; CRISPR/Cas technology; Career Choice; Career Mobility; Cell Line; Cells; Cerebrum; ChIP-seq; Childhood; Chromatin; Clinical; Coculture Techniques; Complement; Coupled; Data; Dependence; Development; Development Plans; Developmental Delay Disorders; Disease; Disease model; Educational workshop; Engineering; Enhancers; Ensure; Environment; Epigenetic Process; Epilepsy; Excision; Fibrinogen; Foundations; Gene Expression; Genes; Genetic; Genetic Models; Genetic Transcription; Genomics; Glass; Grant; Histone Deacetylase; Homeostasis; Human; IL6 gene; IL8 gene; In Vitro; Induced pluripotent stem cell derived neurons; Inflammatory; Inflammatory Response; Institutes; Interferon Type I; Interleukins; Investigation; Journals; Knowledge; Laboratories; Learning; Memory; Mentors; Microglia; Modeling; Molecular; Mus; Nerve Degeneration; Neural Crest; Neuraxis; Neurons; Organoids; Outcome; Outcome Measure; Pathogenesis; Pathogenicity; Pathology; Patients; Phagocytosis; Phenotype; Play; Research; Role; Science; Signal Transduction; Structure; Synapses; Syndrome; System; TNF gene; Technology; Tetracyclines; Time; Training; Training Technics; Transposase; career; career development; cell type; collaborative environment; craniofacial; epigenome; epigenomics; gene environment interaction; genome editing; genome-wide; genome-wide analysis; histone methylation; improved; induced pluripotent stem cell; migration; nerve stem cell; neuroinflammation; neuroregulation; next generation sequencing; novel; overexpression; programs; promoter; research and development; response; stem cell model; stem cells; transcription factor; transcriptome; transcriptome sequencing; transcriptomics

PROJECT SUMMARY Microglia perform essential roles in central nervous system homeostasis with diverse functions including effects on memory and learning as well as refinement of synaptic networks. Emerging evidence indicates that dysregulation of microglia contributes to the pathogenesis of numerous neurodegenerative and neurodevelopmental diseases. Our recent observation that brain environment strongly influences microglial- specific gene expression and regulatory landscapes has implications for understanding the pathogenic response of microglia. The aims in this grant are devised to expand on these data to investigate the transcriptional networks and environmental interactions that influence and maintain the microglial phenotype. Scientific investigations will focus on understanding microglial transcriptional networks and gene- environment interactions using in vitro induced pluripotent stem cell modeling together with cutting edge genomic technologies to provide unbiased molecular outcomes. Specific Aim 1 will investigate the brain- specific signals necessary to maintain the transcriptional network through coculture systems. Not only will this inform gene-environment interactions but will improve iPSC disease modeling to incorporate microglia. Specific Aim 2 will characterize environmentally dependent transcription factors (TFs) through targeted overexpression to understand how these TFs interact with lineage determining TFs and one another to establish functional enhancers and control gene expression. Specific Aim 3 will complement SA2 by reducing expression of an environmentally dependent TF and delineating the microglial contribution to neurodevelopmental pathology. Studies in this grant will utilize the expertise of two excellent and proven mentors, Dr. Christopher Glass who is a leader in genetic and epigenetic analyses and investigating the effect of enhancer landscapes on gene expression and Dr. Fred Gage, a renowned leader in stem cell modeling of disease. The combined training from these two leaders in the field will allow for comprehensive in-depth studies of microglia and contribute to the understanding of how microglia contribute to both the healthy and diseased state as well as improve modeling for neuroinflammatory disease. Together with my mentors and the support of Dr. Blurton- Jones for iPSC microglial differentiations and Dr. Brennand for CRISPR techniques I have a mentoring team that will ensure that I remain on track for career advancement. This mentored award will provide specific training in genomics and epigenetics and allow for a thoughtful combination of stem cell disease modeling with next generation sequencing. In addition, intensive workshops, seminars, journal clubs, and specific laboratory technique training will accompany protected research time. These aims will inform my long-term career objects to model and study pediatric genetic and idiopathic neuroinflammatory disease, and allow for a structured foundation on which to do so. The proposed research and career development plan will benefit greatly from the intellectually rich and collaborative environments at UCSD and the Salk Institute.

项目摘要 小胶质细胞在中枢神经系统稳态中发挥重要作用，具有多种功能 包括对记忆和学习的影响以及突触网络的完善。新兴证据 表明小胶质细胞失调有助于许多神经退行性的发病机理 神经发育疾病。我们最近的观察结果，即大脑环境强烈影响小胶质细胞 特定的基因表达和调节景观对理解致病性具有影响 小胶质细胞的反应。该赠款中的目的是为了扩展这些数据以调查 影响和维持小胶质表型的转录网络和环境相互作用。 科学研究将着重于理解小胶质的转录网络和基因 使用体外诱导的多能干细胞建模与尖端的环境相互作用 基因组技术提供无偏分子结果。具体目标1将研究大脑 通过共培养系统维持转录网络所需的特定信号。不仅会 告知基因环境相互作用，但将改善IPSC疾病建模以纳入小胶质细胞。具体的 AIM 2将通过靶向过表达来表征环境依赖的转录因子（TF） 了解这些TF与谱系如何相互作用，以确定TF和彼此建立功能 增强子和控制基因表达。特定的目标3将通过减少表达来补充SA2 环境依赖的TF，并描绘了对神经发育病理学的小胶质细胞贡献。 该赠款的研究将利用两位优秀而验证的导师克里斯托弗·格拉斯（Christopher Glass）的专业知识 谁是遗传和表观遗传分析的领导者，并研究了增强子景观对 基因表达和弗雷德·盖奇（Fred Gage）博士是疾病干细胞建模领域的著名领导者。组合 该领域的两位领导人的培训将允许对小胶质细胞和 有助于理解小胶质细胞如何对健康状态和患病状态以及 改善神经炎性疾病的建模。与我的导师以及Blurton博士的支持一起 IPSC小胶质分化的琼斯和Brennand博士CRISPR技术我有一个指导团队 这将确保我继续保持职业发展。该指导奖将提供特定的 基因组学和表观遗传学的培训，并允许干细胞疾病建模与 下一代测序。此外，密集的研讨会，研讨会，期刊俱乐部和特定的实验室 技术培训将伴随受保护的研究时间。这些目标将为我的长期职业对象提供信息 建模和研究小儿遗传和特发性神经炎性疾病，并允许结构化 这样做的基础。拟议的研究和职业发展计划将从 UCSD和Salk Institute的智力丰富和协作环境。