Examining Microglial Environmental Dependent Transcriptional Networks

项目摘要 小胶质细胞在维持中枢神经系统稳态中发挥着重要作用，具有多种功能 包括对记忆和学习的影响以及突触网络的细化。新出现的证据 表明小胶质细胞的失调有助于许多神经退行性疾病的发病机制， 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景观对理解致病性 小胶质细胞的反应。该补助金的目的是扩大这些数据，以调查 转录网络和环境相互作用的影响和维持小胶质细胞表型。 科学研究将集中在理解小胶质细胞转录网络和基因- 使用体外诱导多能干细胞建模与尖端技术 基因组技术，以提供公正的分子结果。具体目标1将研究大脑- 通过共培养系统维持转录网络所必需的特定信号。这不仅会 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 目标2将通过靶向过表达来表征环境依赖性转录因子（TF） 为了了解这些TF如何与谱系决定TF相互作用，以及如何相互作用， 增强子和控制基因表达。特异性目标3将通过减少SA 2的表达来补充SA 2。 环境依赖性TF和描绘小胶质细胞对神经发育病理学的贡献。 在这个补助金的研究将利用两个优秀的和成熟的导师，克里斯托弗玻璃博士的专业知识 他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博士，在疾病的干细胞建模的著名领导者。将合并的 来自这两位领域领导者的培训将允许对小胶质细胞进行全面深入的研究， 有助于了解小胶质细胞如何有助于健康和疾病状态，以及 改善神经炎性疾病建模。加上我的导师和布鲁顿博士的支持- Jones负责iPSC小胶质细胞分化，Brennett博士负责CRISPR技术 能确保我的职业发展这一指导奖将提供具体 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，密集的讲习班，研讨会，杂志俱乐部，和具体的实验室 技术培训将伴随受保护的研究时间。这些目标将指导我的长期职业目标 建模和研究儿科遗传性和特发性神经炎性疾病，并允许结构化的 这样做的基础。拟议的研究和职业发展计划将大大受益于 在UCSD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.