Chromatin architecture as a determinant of dendritic cell function

项目摘要/摘要 由表观遗传和遗传机制调节的免疫异质性使我们对免疫的理解变得复杂 监管和疾病。作为哨兵，免疫细胞不断地观察它们的环境，这些信息是 大概是通过表观遗传编码来引导细胞反应。尽管有这样的理解，但人们对此知之甚少。 免疫细胞的表观遗传变异及其与免疫表型异质性的关系。调查是必要的。 为了更好地理解表观遗传变异与免疫应答异质性的关系 个人豁免权。 细胞程序对环境刺激作出反应，对保护性免疫反应是必不可少的。 总体目标：我们的目标是确定DC中的染色质状态是否构成免疫表型的基础 变种。我们将对五个已知免疫的小鼠品系的表观基因组的多样性程度进行表征。 表型变异。我们将绘制染色质可及性图(ATAC-Seq)，并使用一种新的低输入选择组蛋白PTM 染色质定位技术靶下切割释放核酸酶及其与基因的比较 在稳态和免疫刺激后的表达。我们将确定基因或增强子的程度 在DC中启动，以使基因在刺激后快速表达，并识别 驱动直流功能。我们的研究将揭示表观基因组对刺激的反应的动态和静态方面 并确定表观遗传编程在多大程度上是差异激活的基础。我们将开始剖析 免疫细胞遗传变异和表观遗传变异的相互作用及其与免疫表型的关系 异质性。 由于已经克服的技术限制，这些概念在体内来源的DC中的研究很少 由于用于全基因组研究的低投入技术的最新进展。我们已经组建了一个团队，将确保 完成这一项目。我们的合作者带来了生物信息学和表观遗传学技术方面的广泛专业知识，以及 再加上我们在DC生物学方面的丰富经验，创建了一个独特的团队来解决拟议的问题。 影响：在完成这项研究后，我们将提供定义关系的主要概念进展 树突状细胞中染色质状态和功能之间的关系。我们将描述静态与动态方面的 表观基因组对免疫刺激的反应。这些研究还将给我们提供新的见解，以了解 免疫表型变异的染色质构型。这些研究将为未来研究如何 表观基因组的环境扰动--由炎症、营养、肥胖和总体情况引起 健康-与遗传学相互作用，影响免疫细胞编程和免疫反应异质性。

PROJECT SUMMARY/ABSTRACT Immune heterogeneity, mediated by epigenetic and genetic mechanisms, complicates our understanding of immune regulation and disease. As sentinels, immune cells constantly survey their environment, and this information is presumably encoded epigenetically to direct cellular responses. Despite this understanding, little is known about epigenetic variation in immune cells and how it relates to immune phenotype heterogeneity. It is essential to investigate the relationship between epigenetic variation and immune response heterogeneity in order to better understand individual immunity. We will study dendritic cells (DCs), cells of the innate immune system, that engage in distinct cellular programs in response to environmental stimuli and are essential for protective immune responses. OVERALL OBJECTIVES: Our objective is to determine whether chromatin states in DCs underlie immune phenotype variation. We will characterize the degree of diversity in the epigenome of five mouse strains with known immune phenotype variation. We will map chromatin accessibility (ATAC-Seq) and select histone PTMs using a novel low-input chromatin mapping technique Cleavage Under Targets and Release Using Nuclease (CUT&RUN) and compare to gene expression in the steady state and following immune stimulation. We will determine the extent of gene or enhancer priming in DCs that occurs to enable rapid gene expression following stimulation and identify regulatory networks that drive DC function. Our studies will reveal the dynamic and static aspects of the epigenome in response to stimulation and determine the extent to which epigenetic programming underlies differential activation. We will begin to dissect the interplay between genetic and epigenetic variation in immune cells and how it relates to immune phenotype heterogeneity. These concepts have been poorly studied in in vivo-derived DCs due to technical limitations that have been overcome by recent advances in low-input technologies for genome-wide studies. We have assembled a team that will ensure the completion of this project. Our collaborators bring extensive expertise in bioinformatics and epigenetic techniques, and coupled with our extensive experience in DC biology, creates a unique team to address the proposed questions. IMPACT: Upon completion of this research, we will have provided a major conceptual advance defining the relationship between chromatin states and function in DCs. We will have described the static versus dynamic aspects of the epigenome in response to immune stimulation. These studies will also give us new insights into the importance of the chromatin architecture for immune phenotype variation. These studies will lay the foundation for future study of how environmental perturbation of the epigenome—by conditions such as inflammation, nutrition, obesity, and overall health—interacts with genetics to impact immune cell programming and immune response heterogeneity.