Multidimensional Epigenomic Single Cell Analyses

项目总结/摘要 表观基因组是DNA和染色质的化学修饰的集合， 如信使RNA的转录，其进一步充当蛋白质的模板 合成。表观基因组在人类发育过程中的基因调控和 健康和疾病状况。虽然人体内每个细胞的基因组几乎相同， 表观基因组在组织间和组成组织/器官的细胞类型间是高度动态的。的 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， 单个细胞的潜在不同表观基因组。某些生物过程的异质性，如 将体细胞重编程为具有高度医学价值的诱导多能干细胞（iPSC）， 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组分（例如DNA甲基化或 组蛋白修饰）和来自同一样品的转录组。这种多维度的分析， 细胞是非常具有挑战性的，因为大多数当前的方法是一维的。基于高通量测序 该项目将开发分析多种表观基因组成分的方法，包括DNA 甲基化组和染色质可及性，以及单个人类细胞的转录组。这些方法将 最初使用培养的人类细胞开发，后来适应于原代人类组织。作为分析模型 发展，该方法将应用于研究单细胞表观基因组多样性在体细胞 这是一个产生iPSCs的重编程过程。如果开发成功，这些方法将大大促进 在复杂的人体组织和异质性疾病中， 癌

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.