Genetics Distinct Epigenomic Features in End-Stage Failing Human Hearts Mehregan Movassagh, MPhil, PhD; Mun-Kit Choy, PhD; David A. Knowles, MA, MSc, MEng; Lina Cordeddu, PhD; Syed Haider, MSc; Thomas Down, PhD; Lee Siggens, BSc, MPhil; Ana Vujic, MSc, MPhil; Ilenia Simeoni, PhD; Chris Penkett, PhD; Martin Goddard, MBBCh, FRCS, FRCPath; Pietro Lio, PhD; Martin R. Bennett, MA, PhD, FRCP, FMedSci; Roger S.-Y. Foo, MD, MBBS, MRCP Background—The epigenome refers to marks on the genome, including DNA methylation and histone modifications, that regulate the expression of underlying genes. A consistent profile of gene expression changes in end-stage cardiomy- opathy led us to hypothesize that distinct global patterns of the epigenome may also exist. Methods and Results—We constructed genome-wide maps of DNA methylation and histone-3 lysine-36 trimethylation (H3K36me3) enrichment for cardiomyopathic and normal human hearts. More than 506 Mb sequences per library were generated by high-throughput sequencing, allowing us to assign methylation scores to 28 million CG dinucleotides in the human genome. DNA methylation was significantly different in promoter CpG islands, intragenic CpG islands, gene bodies, and H3K36me3-enriched regions of the genome. DNA methylation differences were present in promoters of upregulated genes but not downregulated genes. H3K36me3 enrichment itself was also significantly different in coding regions of the genome. Specifically, abundance of RNA transcripts encoded by the DUX4 locus correlated to differential DNA methylation and H3K36me3 enrichment. In vitro, Dux gene expression was responsive to a specific inhibitor of DNA methyltransferase, and Dux siRNA knockdown led to reduced cell viability. Conclusions—Distinct epigenomic patterns exist in important DNA elements of the cardiac genome in human end-stage cardiomyopathy. The epigenome may control the expression of local or distal genes with critical functions in myocardial stress response. If epigenomic patterns track with disease progression, assays for the epigenome may be useful for assessing prognosis in heart failure. Further studies are needed to determine whether and how the epigenome contributes to the development of cardiomyopathy. (Circulation. 2011;124:2411-2422.) Key Words: genes genome-wide analysis genomics heart failure A ltered gene expression is a characteristic hallmark of heart failure. 1–4 Moreover, analysis of the transcrip- tome in human cardiomyopathy shows that altered gene expression may be used to predict disease severity. 5 Although gene expression changes can be explained by pathways that link to the assembly of cardiac transcription factors at gene promoters during myocardial stress, 6 our better understanding of epigenomics 7 now allows us to determine whether epigenomic changes also occur in heart failure. Clinical Perspective on p 2422 The epigenome refers to marks on the genome, including DNA methylation and histone modification. These marks do not change the underlying genetic sequence but regulate how underlying genes are expressed. 7 Unlike the genome, which is largely stable, the epigenome is dynamic and allows organisms to respond and adapt to environmental cues. The environment therefore promotes epigenomic changes with consequent plasticity in gene expression and phenotype. 8 Strong evidence indicates that drifts in DNA methylation occur with increasing age. 9,10 In addition, the recent discovery of active DNA demethylation in postmitotic cells 11 and rapid and dynamic DNA methylation/demethylation in vivo 12,13 further challenges the conventional view that DNA methyl- ation is a stable epigenetic mark. Dynamic changes in DNA methylation may therefore link the environment to disease pathogenesis. 8 The role of DNA methylation has been proven in cancer 14 and implicated in other complex diseases, includ- Received January 20, 2011; accepted September 14, 2011. From the Division of Cardiovascular Medicine (M.M., M.-K.C., L.C., L.S., A.V., M.R.B., R.S.-Y.F.), Department of Engineering (D.A.K.), Computer Laboratory (S.H., P.L.), Gurdon Institute and Department of Genetics (T.D.), and East Anglian Sequencing and Bioinformatics Hub and Department of Medical Genetics (C.P., I.S.), University of Cambridge; and Department of Histopathology, Papworth Hospital, Papworth Everard, Cambridge, UK (M.G.). The online-only Data Supplement is available with this article at http://circ.ahajournals.org/lookup/suppl/doi:10.1161/CIRCULATIONAHA.111. 040071/-/DC1. Correspondence to Roger Foo, MD, MBBS, MRCP, Division of Cardiovascular Medicine, University of Cambridge, Addenbrooke’s Centre for Clinical Investigation, Level 6, Hills Rd, Cambridge, CB2 0QQ UK. E-mail [email protected] © 2011 American Heart Association, Inc. Circulation is available at http://circ.ahajournals.org DOI: 10.1161/CIRCULATIONAHA.111.040071 2411 by guest on May 8, 2018 http://circ.ahajournals.org/ Downloaded from by guest on May 8, 2018 http://circ.ahajournals.org/ Downloaded from by guest on May 8, 2018 http://circ.ahajournals.org/ Downloaded from by guest on May 8, 2018 http://circ.ahajournals.org/ Downloaded from by guest on May 8, 2018 http://circ.ahajournals.org/ Downloaded from by guest on May 8, 2018 http://circ.ahajournals.org/ Downloaded from by guest on May 8, 2018 http://circ.ahajournals.org/ Downloaded from by guest on May 8, 2018 http://circ.ahajournals.org/ Downloaded from by guest on May 8, 2018 http://circ.ahajournals.org/ Downloaded from by guest on May 8, 2018 http://circ.ahajournals.org/ Downloaded from by guest on May 8, 2018 http://circ.ahajournals.org/ Downloaded from by guest on May 8, 2018 http://circ.ahajournals.org/ Downloaded from by guest on May 8, 2018 http://circ.ahajournals.org/ Downloaded from by 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