High resolution genomic and epigenomic mapping of the human salivary gland

项目摘要 分化程序和正常组织的机制和分子检测 唾液腺（SG）的稳态是一个重要的研究领域，因为改变的SG功能是 与多种人类疾病状况相关，包括干燥综合征、癌症和由于 癌症化疗/放射治疗。这就需要深入研究复杂的生态系统， SG由各种上皮和非上皮细胞群组成，这些细胞群相互协作， 促进SG功能。转录和表观基因组基因调控的潜在分子电路 控制SG生物学的机制，特别是当它涉及体内环境中的单个细胞类型时， 然而，人类SG并不被很好地理解，因此存在显着的知识差距。缺乏 这种知识阻碍了对细胞命运和谱系选择、细胞间遗传和细胞间分化的原理的更好理解。 这些信息传递和转录调节过程是指导有效治疗所需的 人类疾病的干预。我们的中心假设是SG转录组的建立， 表观基因组和基因调控网络是一个动态的过程，是相互作用的结果 细胞内信号通路和每种细胞类型的潜在硬连线基因组信息之间的联系。 为了检验这一假设，提出了三个具体目标。目的1是产生单细胞RNA测序 （scRNA-seq）和来自成年雄性和雌性SMG的相同细胞的scATAC-seq数据。Aim的目标 2是使用计算工具来定义细胞命运轨迹和细胞间通信系统， 在SG中操作，并确定决定细胞命运的关键转录调节因子。第三，3D 将通过HiChIP建立人SG的染色质状态和增强子-启动子连接体图谱 实验这些数据将有助于建立非编码遗传变异与 与疾病相关的疾病相关基因，如主要折磨人的干燥综合征， SG.这项工作具有高度创新性和重要性，因为我们建议使用尖端技术， 研究基因调控的基本转录和表观基因组机制的复杂工具 和信号传导途径。从长远来看，这些知识将大大推动 对SG生物学的基本了解，预计将对治疗 SG的复杂遗传疾病。

PROJECT SUMMARY A mechanistic and molecular examination of the differentiation programs and normal tissue homeostasis of the salivary glands (SG) is an important area of research since altered SG function is associated with multiple human disease conditions including Sjogren syndrome, cancer, and complications due to cancer chemo/radiation therapy. This necessitates an in-depth investigation of the complex ecosystem of SG that consists of a variety of epithelial and non-epithelial cell populations that cooperatively interact to facilitate SG function. The underlying molecular circuitry of the transcriptional and epigenomic gene-regulatory mechanisms that control SG biology, particularly as it pertains to individual cell types in the in vivo context of the human SG, however, is not very well-understood and thus presents a significant knowledge gap. Lack of this knowledge prevents a better understanding of principles of cell fate and lineage choices, cell-to-cell communication and transcriptional regulatory processes that are needed for guiding effective therapeutic interventions of human diseases. Our central hypothesis is that the establishment of the SG transcriptome, epigenome and gene regulatory networks is a dynamic process that results from reciprocal interactions between intracellular signaling pathways and the underlying hardwired genomic information of each cell-type. To test this hypothesis, three specific aims are proposed. Aim 1 is to generate single-cell RNA-sequencing (scRNA-seq), and scATAC-seq data from the same cells of the adult male and female SMG. The goals of Aim 2 are to use computational tools to define the cell fate trajectories and cell-to-cell communication systems that operate in the SG and identify crucial transcriptional regulators that define cell fate. Finally in Aim 3, the 3D chromatin state and the enhancer-promoter connectome map of the human SG will be established by HiChIP experiments. Such data will enable the establishment of the link between non-coding genetic variants and disease-associated genes that are relevant for disease such as Sjogren’s syndrome that primarily afflict the SG. This work is highly innovative and significant because our proposed use of cutting-edge technologies and sophisticated tools to examine fundamental transcriptional and epigenomic mechanisms of gene regulation and signaling pathways at a single cell resolution. Long term, such knowledge will substantially advance the fundamental understanding of SG biology and is anticipated to have a long-term impact on the treatment of complex genetic diseases of the SG.