Genetic analysis of the cardiac methylome at single nucleotide resolution in a model of human cardiovascular disease.

Epigenetic marks such as cytosine methylation are important determinants of cellular and whole-body phenotypes. However, the extent of, and reasons for inter-individual differences in cytosine methylation, and their association with phenotypic variation are poorly characterised. Here we present the...

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Main Authors: Michelle D Johnson, Michael Mueller, Martyna Adamowicz-Brice, Melissa J Collins, Pascal Gellert, Klio Maratou, Prashant K Srivastava, Maxime Rotival, Shahena Butt, Laurence Game, Santosh S Atanur, Nicholas Silver, Penny J Norsworthy, Sarah R Langley, Enrico Petretto, Michal Pravenec, Timothy J Aitman
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2014-12-01
Series:PLoS Genetics
Online Access:https://doi.org/10.1371/journal.pgen.1004813
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author Michelle D Johnson
Michael Mueller
Martyna Adamowicz-Brice
Melissa J Collins
Pascal Gellert
Klio Maratou
Prashant K Srivastava
Maxime Rotival
Shahena Butt
Laurence Game
Santosh S Atanur
Nicholas Silver
Penny J Norsworthy
Sarah R Langley
Enrico Petretto
Michal Pravenec
Timothy J Aitman
author_facet Michelle D Johnson
Michael Mueller
Martyna Adamowicz-Brice
Melissa J Collins
Pascal Gellert
Klio Maratou
Prashant K Srivastava
Maxime Rotival
Shahena Butt
Laurence Game
Santosh S Atanur
Nicholas Silver
Penny J Norsworthy
Sarah R Langley
Enrico Petretto
Michal Pravenec
Timothy J Aitman
author_sort Michelle D Johnson
collection DOAJ
description Epigenetic marks such as cytosine methylation are important determinants of cellular and whole-body phenotypes. However, the extent of, and reasons for inter-individual differences in cytosine methylation, and their association with phenotypic variation are poorly characterised. Here we present the first genome-wide study of cytosine methylation at single-nucleotide resolution in an animal model of human disease. We used whole-genome bisulfite sequencing in the spontaneously hypertensive rat (SHR), a model of cardiovascular disease, and the Brown Norway (BN) control strain, to define the genetic architecture of cytosine methylation in the mammalian heart and to test for association between methylation and pathophysiological phenotypes. Analysis of 10.6 million CpG dinucleotides identified 77,088 CpGs that were differentially methylated between the strains. In F1 hybrids we found 38,152 CpGs showing allele-specific methylation and 145 regions with parent-of-origin effects on methylation. Cis-linkage explained almost 60% of inter-strain variation in methylation at a subset of loci tested for linkage in a panel of recombinant inbred (RI) strains. Methylation analysis in isolated cardiomyocytes showed that in the majority of cases methylation differences in cardiomyocytes and non-cardiomyocytes were strain-dependent, confirming a strong genetic component for cytosine methylation. We observed preferential nucleotide usage associated with increased and decreased methylation that is remarkably conserved across species, suggesting a common mechanism for germline control of inter-individual variation in CpG methylation. In the RI strain panel, we found significant correlation of CpG methylation and levels of serum chromogranin B (CgB), a proposed biomarker of heart failure, which is evidence for a link between germline DNA sequence variation, CpG methylation differences and pathophysiological phenotypes in the SHR strain. Together, these results will stimulate further investigation of the molecular basis of locally regulated variation in CpG methylation and provide a starting point for understanding the relationship between the genetic control of CpG methylation and disease phenotypes.
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spelling doaj-art-05edfc5efe3442ebbf5bdb0c4e3f8e9f2025-08-20T02:22:45ZengPublic Library of Science (PLoS)PLoS Genetics1553-73901553-74042014-12-011012e100481310.1371/journal.pgen.1004813Genetic analysis of the cardiac methylome at single nucleotide resolution in a model of human cardiovascular disease.Michelle D JohnsonMichael MuellerMartyna Adamowicz-BriceMelissa J CollinsPascal GellertKlio MaratouPrashant K SrivastavaMaxime RotivalShahena ButtLaurence GameSantosh S AtanurNicholas SilverPenny J NorsworthySarah R LangleyEnrico PetrettoMichal PravenecTimothy J AitmanEpigenetic marks such as cytosine methylation are important determinants of cellular and whole-body phenotypes. However, the extent of, and reasons for inter-individual differences in cytosine methylation, and their association with phenotypic variation are poorly characterised. Here we present the first genome-wide study of cytosine methylation at single-nucleotide resolution in an animal model of human disease. We used whole-genome bisulfite sequencing in the spontaneously hypertensive rat (SHR), a model of cardiovascular disease, and the Brown Norway (BN) control strain, to define the genetic architecture of cytosine methylation in the mammalian heart and to test for association between methylation and pathophysiological phenotypes. Analysis of 10.6 million CpG dinucleotides identified 77,088 CpGs that were differentially methylated between the strains. In F1 hybrids we found 38,152 CpGs showing allele-specific methylation and 145 regions with parent-of-origin effects on methylation. Cis-linkage explained almost 60% of inter-strain variation in methylation at a subset of loci tested for linkage in a panel of recombinant inbred (RI) strains. Methylation analysis in isolated cardiomyocytes showed that in the majority of cases methylation differences in cardiomyocytes and non-cardiomyocytes were strain-dependent, confirming a strong genetic component for cytosine methylation. We observed preferential nucleotide usage associated with increased and decreased methylation that is remarkably conserved across species, suggesting a common mechanism for germline control of inter-individual variation in CpG methylation. In the RI strain panel, we found significant correlation of CpG methylation and levels of serum chromogranin B (CgB), a proposed biomarker of heart failure, which is evidence for a link between germline DNA sequence variation, CpG methylation differences and pathophysiological phenotypes in the SHR strain. Together, these results will stimulate further investigation of the molecular basis of locally regulated variation in CpG methylation and provide a starting point for understanding the relationship between the genetic control of CpG methylation and disease phenotypes.https://doi.org/10.1371/journal.pgen.1004813
spellingShingle Michelle D Johnson
Michael Mueller
Martyna Adamowicz-Brice
Melissa J Collins
Pascal Gellert
Klio Maratou
Prashant K Srivastava
Maxime Rotival
Shahena Butt
Laurence Game
Santosh S Atanur
Nicholas Silver
Penny J Norsworthy
Sarah R Langley
Enrico Petretto
Michal Pravenec
Timothy J Aitman
Genetic analysis of the cardiac methylome at single nucleotide resolution in a model of human cardiovascular disease.
PLoS Genetics
title Genetic analysis of the cardiac methylome at single nucleotide resolution in a model of human cardiovascular disease.
title_full Genetic analysis of the cardiac methylome at single nucleotide resolution in a model of human cardiovascular disease.
title_fullStr Genetic analysis of the cardiac methylome at single nucleotide resolution in a model of human cardiovascular disease.
title_full_unstemmed Genetic analysis of the cardiac methylome at single nucleotide resolution in a model of human cardiovascular disease.
title_short Genetic analysis of the cardiac methylome at single nucleotide resolution in a model of human cardiovascular disease.
title_sort genetic analysis of the cardiac methylome at single nucleotide resolution in a model of human cardiovascular disease
url https://doi.org/10.1371/journal.pgen.1004813
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