Faculty, Staff and Student Publications
Language
English
Publication Date
8-28-2025
Journal
Cardiovascular Research
DOI
10.1093/cvr/cvae063
PMID
38577741
PMCID
PMC12641529
PubMedCentral® Posted Date
4-5-2024
PubMedCentral® Full Text Version
Post-print
Abstract
Aims: An intrinsic feature of gene transcription is the formation of DNA superhelices near the transcription bubble, which are resolved upon induction of transient double-stranded breaks (DSBs) by topoisomerases. Unrepaired DSBs are pathogenic as they lead to cell cycle arrest, senescence, inflammation, and organ dysfunction. We posit that DSBs would be more prevalent at the genomic sites that are associated with gene expression. The objectives were to identify and characterize genome-wide DSBs at the nucleotide resolution and determine the association of DSBs with transcription in cardiac myocytes.
Methods and results: We identified the genome-wide DSBs in ∼1 million cardiac myocytes per heart in three wild-type and three myocyte-specific LMNA-deficient (Myh6-Cre:LmnaF/F) mice by END-Sequencing. The prevalence of DSBs was 0.8% and 2.2% in the wild-type and Myh6-Cre:LmnaF/F myocytes, respectively. The END-Seq signals were enriched for 8 and 6764 DSBs in the wild-type and Myh6-Cre:LmnaF/F myocytes, respectively (q < 0.05). The DSBs were preferentially localized to the gene regions, transcription initiation sites, cardiac transcription factor motifs, and the G quadruplex forming structures. Because LMNA regulates transcription through the lamin-associated domains (LADs), we defined the LADs in cardiac myocytes by a Cleavage Under Targets & Release Using Nuclease (CUT&RUN) assay (N = 5). On average there were 818 LADs per myocyte. Constitutive LADs (cLADs), defined as LADs that were shared by at least three genomes (N = 2572), comprised about a third of the mouse cardiac myocyte genomes. Transcript levels of the protein-coding genes located at the cLADs (N = 3975) were ∼16-fold lower than those at the non-LAD regions (N = ∼17 778). The prevalence of DSBs was higher in the non-LAD as compared to the cLAD regions. Likewise, DSBs were more common in the loss-of-LAD regions, defined as the genomic regions in the Myh6-Cre:LmnaF/F that were juxtaposed to the LAD regions in the wild-type myocytes.
Conclusion: To our knowledge, this is the first identification of the DSBs, at the nucleotide resolution in the cardiovascular system. The prevalence of DSBs was higher in the genomic regions associated with transcription. Because transcription is pervasive, DSBs are expected to be common and pathogenic in various states and aging.
Keywords
Animals, Myocytes, Cardiac, Lamin Type A, Transcription, Genetic, DNA Breaks, Double-Stranded, Aging, Disease Models, Animal, Cardiomyopathies, Mice, Knockout, Male, Age Factors, Gene Expression Regulation, Mice, Inbred C57BL, Mice, Mice, Transgenic, Genomic instability, Double-stranded DNA breaks, Transcriptional stress, Lamin A, Heart failure, Aging
Published Open-Access
yes
Recommended Citation
Cathcart, Benjamin; Cheedipudi, Sirisha M; Rouhi, Leila; et al., "DNA Double-Stranded Breaks, a Hallmark of Aging, Defined at the Nucleotide Resolution, Are Increased and Associated With Transcription in the Cardiac Myocytes in LMNA-Cardiomyopathy" (2025). Faculty, Staff and Student Publications. 799.
https://digitalcommons.library.tmc.edu/uthshis_docs/799
Graphical Abstract