Phenotypic switching induced by damaged matrix is associated with DNA methyltransferase 3A (DNMT3A) activity and nuclear localization in smooth muscle cells (SMC).

TitlePhenotypic switching induced by damaged matrix is associated with DNA methyltransferase 3A (DNMT3A) activity and nuclear localization in smooth muscle cells (SMC).
Publication TypeJournal Article
Year of Publication2013
AuthorsJiang, J-X., K. J. Aitken, C. Sotiropoulos, C. Sotiropolous, T. Kirwan, T. Panchal, N. Zhang, S. Pu, S. J. Wodak, C. Tolg, and D. J. B├Ągli
JournalPLoS One
Volume8
Issue8
Paginatione69089
Date Published2013
ISSN1932-6203
KeywordsActive Transport, Cell Nucleus, Animals, Cell Count, Cell Dedifferentiation, Cell Nucleus, Cell Proliferation, Collagen, DNA (Cytosine-5-)-Methyltransferase, DNA Methylation, Extracellular Matrix, Gene Expression Regulation, Enzymologic, Humans, Intracellular Space, Mitosis, Myocytes, Smooth Muscle, Phenotype, Protein Biosynthesis, Protein Denaturation, Protein Transport, Rats, Signal Transduction, Time Factors, Transcription, Genetic
Abstract

Extracellular matrix changes are often crucial inciting events for fibroproliferative disease. Epigenetic changes, specifically DNA methylation, are critical factors underlying differentiated phenotypes. We examined the dependency of matrix-induced fibroproliferation and SMC phenotype on DNA methyltransferases. The cooperativity of matrix with growth factors, cell density and hypoxia was also examined. Primary rat visceral SMC of early passage (0-2) were plated on native collagen or damaged/heat-denatured collagen. Hypoxia was induced with 3% O2 (balanced 5% CO2 and 95% N2) over 48 hours. Inhibitors were applied 2-3 hours after cells were plated on matrix, or immediately before hypoxia. Cells were fixed and stained for DNMT3A and smooth muscle actin (SMA) or smooth muscle myosin heavy chain. Illumina 450 K array of CpG sites was performed on bisulfite-converted DNA from smooth muscle cells on damaged matrix vs native collagen. Matrix exquisitely regulates DNMT3A localization and expression, and influences differentiation in SMCs exposed to denatured matrix +/- hypoxia. Analysis of DNA methylation signatures showed that Matrix caused significant DNA methylation alterations in a discrete number of CpG sites proximal to genes related to SMC differentiation. Matrix has a profound effect on the regulation of SMC phenotype, which is associated with altered expression, localization of DNMTs and discrete changes DNA methylation.

DOI10.1371/journal.pone.0069089
Alternate JournalPLoS ONE
PubMed ID24282625
PubMed Central IDPMC3735580
Grant List / / Canadian Institutes of Health Research / Canada
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