Multidimensional Epigenomic Single Cell Analyses

项目摘要/摘要 表观基因组是调节基因组的dna和染色质的化学修饰的集合。 信使RNA的转录等活动，它进一步充当蛋白质的模板 综合。表观基因组在人类发育过程中的基因调控中起着重要作用。 健康状况和疾病状况。尽管人体内每个细胞的基因组几乎完全相同， 表观基因组在组织之间和组成组织/器官的细胞类型之间是高度动态的。这个 目前对表观基因组的了解主要建立在对人体组织的分析上，而没有区分 单个细胞的潜在不同的表观基因组。某些生物过程的异质性，例如 将体细胞重新编程为具有高度医学价值的诱导多能干细胞(IPSCs)，也 需要分析单细胞的表观基因组和转录组的能力。一种强大的方法 研究表观基因组调控是为了分析表观基因组的多个成分(例如DNA甲基化或 组蛋白修饰)和转录组。这种单一的多维分析 细胞是极具挑战性的，因为目前的大多数方法都是一维的。基于高通量测序的 该项目将开发分析多种表观基因组成分的方法，包括DNA 甲基组和染色质的可及性，以及单个人类细胞的转录组。这些方法将是 最初是利用培养的人类细胞开发的，后来适应于原始的人体组织。作为化验的模型 发展，这些方法将被应用于体细胞单细胞表观基因组多样性的研究 对生成IPSC的过程进行重新编程。如果开发成功，这些方法将极大地促进 复杂人体组织和异质疾病中不同细胞类型的表观基因组学研究 癌症。

Project Summary/Abstract The epigenome is the ensemble of chemical modifications of DNA and chromatin that modulates genomic activities such as the transcription of messenger RNA, which further serves as the template of protein syntheses. The epigenome plays instrumental roles in gene regulation during human development and in healthy as well as disease conditions. Although the genome is nearly identical in each cell in the human body, epigenomes are highly dynamic across tissues and between cell types composing the tissues/organs. The current knowledge of the epigenome is largely built on analyses of human tissues without distinguishing the potential distinct epigenomes of individual cells. The heterogeneity of certain biological processes, such as the reprogramming of somatic cells to the highly medically valuable induced pluripotent stem cells (iPSCs), also requires the capability of analyzing the epigenome and transcriptome of single cells. A powerful approach for studying epigenomic regulation is to profile multiple components of the epigenome (e.g. DNA methylation or histone modifications) and the transcriptome from the same sample. Such multi-dimensional analysis of single cells is highly challenging since most current methods are uni-dimensional. High-throughput sequencing based methods will be developed by the project to analyze multiple epigenomic components, including the DNA methylome and chromatin accessibility, and the transcriptome of single human cells. These methods will be first developed using cultured human cells and later adapted to primary human tissues. As a model for assay development, the methods will be applied to study single cell epigenomic diversity during the somatic cell reprogramming process that generates iPSCs. If successfully developed, these methods will greatly facilitate epigenomic studies of diverse cell types in complex human tissues and in heterogeneous diseases such as cancer.