title: Genome-wide detection of DNase I hypersensitive sites in single cells and FFPE tissue samples.
abstract: DNase I hypersensitive sites (DHSs) provide important information on the presence of transcriptional regulatory elements and the state of chromatin in mammalian cells. Conventional DNase sequencing (DNase-seq) for genome-wide DHSs profiling is limited by the requirement of millions of cells. Here we report an ultrasensitive strategy, called single-cell DNase sequencing (scDNase-seq) for detection of genome-wide DHSs in single cells. We show that DHS patterns at the single-cell level are highly reproducible among individual cells. Among different single cells, highly expressed gene promoters and enhancers associated with multiple active histone modifications display constitutive DHS whereas chromatin regions with fewer histone modifications exhibit high variation of DHS. Furthermore, the single-cell DHSs predict enhancers that regulate cell-specific gene expression programs and the cell-to-cell variations of DHS are predictive of gene expression. Finally, we apply scDNase-seq to pools of tumour cells and pools of normal cells, dissected from formalin-fixed paraffin-embedded tissue slides from patients with thyroid cancer, and detect thousands of tumour-specific DHSs. Many of these DHSs are associated with promoters and enhancers critically involved in cancer development. Analysis of the DHS sequences uncovers one mutation (chr18: 52417839G>C) in the tumour cells of a patient with follicular thyroid carcinoma, which affects the binding of the tumour suppressor protein p53 and correlates with decreased expression of its target gene TXNL1. In conclusion, scDNase-seq can reliably detect DHSs in single cells, greatly extending the range of applications of DHS analysis both for basic and for translational research, and may provide critical information for personalized medicine.
authors.affiliation: Systems Biology Center, Division of Intramural Research, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: Wenfei
authors.initials: W
authors.lastname: Jin
authors.affiliation: Systems Biology Center, Division of Intramural Research, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: Qingsong
authors.initials: Q
authors.lastname: Tang
authors.affiliation: Department of Pathology, Yale University School of Medicine, New Haven, Connecticut 06520, USA.
authors.forename: Mimi
authors.initials: M
authors.lastname: Wan
authors.affiliation: Systems Biology Center, Division of Intramural Research, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: Kairong
authors.initials: K
authors.lastname: Cui
authors.affiliation: Systems Biology Center, Division of Intramural Research, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.|Institute of Immunology, Third Military Medical University of the People's Liberation Army, Chongqing 400038, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Systems Biology Center, Division of Intramural Research, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.|College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi 712100, China.
authors.forename: Gang
authors.initials: G
authors.lastname: Ren
authors.affiliation: Institute of Immunology, Third Military Medical University of the People's Liberation Army, Chongqing 400038, China.
authors.forename: Bing
authors.initials: B
authors.lastname: Ni
authors.affiliation: Department of Pathology, Yale University School of Medicine, New Haven, Connecticut 06520, USA.
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Sklar
authors.affiliation: Computational Biology Branch, National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: Teresa M
authors.initials: TM
authors.lastname: Przytycka
authors.affiliation: Hematology Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: Richard
authors.initials: R
authors.lastname: Childs
authors.affiliation: Laboratory of Pathology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: David
authors.initials: D
authors.lastname: Levens
authors.affiliation: Systems Biology Center, Division of Intramural Research, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: Keji
authors.initials: K
authors.lastname: Zhao
mesh_terms: D018263:Adenocarcinoma, Follicular / Q000235:genetics / Q000473:pathology; D000818:Animals; D002843:Chromatin / Q000235:genetics* / Q000378:metabolism*; D003850:Deoxyribonuclease I / Q000378:metabolism*; D004742:Enhancer Elements, Genetic / Q000235:genetics; D005557:Formaldehyde; D020869:Gene Expression Profiling; D016678:Genome / Q000235:genetics*; D006657:Histones / Q000378:metabolism; D006801:Humans; D051379:Mice; D009154:Mutation / Q000235:genetics; D041681:NIH 3T3 Cells; D016612:Paraffin Embedding; D011401:Promoter Regions, Genetic / Q000235:genetics; D015203:Reproducibility of Results; D059010:Single-Cell Analysis / Q000379:methods*; D013879:Thioredoxins / Q000235:genetics; D013964:Thyroid Neoplasms / Q000235:genetics / Q000473:pathology; D016707:Tissue Fixation; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
chemical_list: D002843:Chromatin; D006657:Histones; C495901:TP53 protein, human; C504766:TXNL1 protein, human; D016159:Tumor Suppressor Protein p53; D005557:Formaldehyde; D013879:Thioredoxins; D003850:Deoxyribonuclease I