Examining Microglial Environmental Dependent Transcriptional Networks

项目总结 小胶质细胞在中枢神经系统内稳中起重要作用，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。新出现的证据 提示小胶质细胞调节失调参与了多种神经退行性变和 神经发育疾病。我们最近观察到，大脑环境强烈影响小胶质细胞- 特定的基因表达和调控格局对于理解致病机理具有重要意义。 小胶质细胞的反应。这项拨款的目的是扩大这些数据，以调查 影响和维持小胶质细胞表型的转录网络和环境相互作用。 科学研究将侧重于了解小胶质细胞转录网络和基因- 使用体外诱导多能干细胞模型和尖端技术的环境交互作用 提供公正的分子结果的基因组技术。特定目标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。这不仅会让你 告知基因-环境相互作用，但将改进IPSC疾病建模，以纳入小胶质细胞。特定的 目标2将通过定向过表达来表征环境依赖的转录因子 为了了解这些功能因子如何与决定血统的功能因子相互作用，以及如何相互作用来建立功能 增强剂和控制基因表达。特异靶蛋白3将通过减少An的表达来补充SA2 环境依赖性转铁蛋白和描绘小胶质细胞对神经发育病理学的贡献。 这笔赠款的研究将利用两位优秀的、久经考验的导师克里斯托弗·格拉斯博士的专业知识 世卫组织在遗传和表观遗传学分析以及研究增强剂景观的影响方面处于领先地位 基因表达和弗雷德·盖奇博士，他是疾病干细胞建模的著名领导者。合并后的 来自这两位领域领导者的培训将允许全面深入地研究小胶质细胞和 有助于理解小胶质细胞如何对健康和疾病状态以及 改进神经炎症性疾病的建模。在我的导师和布鲁顿博士的支持下- 琼斯负责IPSC小胶质细胞分化，布伦南博士负责CRISPR技术，我有一个指导团队 这将确保我继续保持在职业发展的轨道上。这一指导奖将提供具体的 基因组学和表观遗传学方面的培训，并允许将干细胞疾病建模与 下一代测序。此外，密集的研讨会、研讨会、杂志俱乐部和特定的实验室 技术培训将伴随着受保护的研究时间。这些目标将决定我的长期职业目标 模拟和研究儿童遗传性和特发性神经炎性疾病，并允许一个结构化的 这样做所依据的基础。拟议的研究和职业发展计划将大大受益于 加州大学圣迭戈分校和索尔克研究所的智力丰富和协作环境。

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.