Folate deficiency promotes differentiation of vascular smooth muscle cells without affecting the methylation status of regulated genes

Andreas F. Kolb* (Corresponding Author), Linda Petrie, Claus D. Mayer, Lynn Pirie, Susan J. Duthie

*Corresponding author for this work

Research output: Contribution to journalArticle

Abstract

Elevated serum homocysteine, an intermediate of cellular one-carbon metabolism, is an independent risk factor for cardiovascular disease (CVD). Folate deficiency increases serum homocysteine and may contribute to CVD progression. Vascular smooth muscle cells regulate vascular contractility, but also contribute to repair processes in response to vascular injury. Nutritional deficiencies, like folate deficiency, are thought to impact on this phenotypic plasticity, possibly by epigenetic mechanisms. We have investigated the effect of folate deficiency on vascular smooth muscle cells in two cell culture systems representing early and late stages of smooth muscle cells differentiation. We find that folate deficiency promotes differentiation towards a more contractile phenotype as indicated by increased expression of respective marker genes. However, microarray analysis identified markers of striated muscle as the predominant gene expression change elicited by folate deficiency. These changes are not merely a reflection of cell cycle arrest, as foetal calf serum restriction or iron deficiency do not replicate the gene expression changes observed in response to folate deficiency. Folate deficiency only has a marginal effect on global DNA methylation. DNA methylation of CpG islands associated with genes regulated by folate deficiency remains unaffected. This supports our earlier findings in a mouse model system which also did not show any changes in global DNA methylation in response to folate and vitamin B6/B12 deficiency. These data suggest that folate deficiency enhances the expression of smooth muscle marker gene expression, promotes a shift towards a skeletal muscle phenotype, and does not regulate gene expression via DNA methylation.
Original languageEnglish
Pages (from-to)2769-2795
Number of pages7
JournalBiochemical Journal
Volume476
Issue number19
Early online date17 Sep 2019
DOIs
Publication statusPublished - 2019

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Methylation
Vascular Smooth Muscle
Folic Acid
Smooth Muscle Myocytes
Muscle
Genes
Cells
DNA Methylation
Gene expression
Gene Expression
Homocysteine
Cardiovascular Diseases
Serum
Vitamin B 6 Deficiency
Phenotype
Vitamin B 12 Deficiency
CpG Islands
Vitamin B 6
Striated Muscle
Vascular System Injuries

Keywords

  • B-vitamins
  • folate
  • vascular smooth muscle cells
  • DNA methylation
  • gene regulation
  • FOLIC-ACID FORTIFICATION
  • HYPERHOMOCYSTEINEMIA
  • VITAMINS-B
  • DNA METHYLATION
  • MICE DEFICIENT
  • ATHEROSCLEROSIS
  • SUPPLEMENTATION
  • NUTRITION
  • EXPRESSION
  • PLASMA HOMOCYSTEINE

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry
  • Cell Biology

Cite this

Folate deficiency promotes differentiation of vascular smooth muscle cells without affecting the methylation status of regulated genes. / Kolb, Andreas F. (Corresponding Author); Petrie, Linda; Mayer, Claus D.; Pirie, Lynn; Duthie, Susan J.

In: Biochemical Journal, Vol. 476, No. 19, 2019, p. 2769-2795.

Research output: Contribution to journalArticle

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abstract = "Elevated serum homocysteine, an intermediate of cellular one-carbon metabolism, is an independent risk factor for cardiovascular disease (CVD). Folate deficiency increases serum homocysteine and may contribute to CVD progression. Vascular smooth muscle cells regulate vascular contractility, but also contribute to repair processes in response to vascular injury. Nutritional deficiencies, like folate deficiency, are thought to impact on this phenotypic plasticity, possibly by epigenetic mechanisms. We have investigated the effect of folate deficiency on vascular smooth muscle cells in two cell culture systems representing early and late stages of smooth muscle cells differentiation. We find that folate deficiency promotes differentiation towards a more contractile phenotype as indicated by increased expression of respective marker genes. However, microarray analysis identified markers of striated muscle as the predominant gene expression change elicited by folate deficiency. These changes are not merely a reflection of cell cycle arrest, as foetal calf serum restriction or iron deficiency do not replicate the gene expression changes observed in response to folate deficiency. Folate deficiency only has a marginal effect on global DNA methylation. DNA methylation of CpG islands associated with genes regulated by folate deficiency remains unaffected. This supports our earlier findings in a mouse model system which also did not show any changes in global DNA methylation in response to folate and vitamin B6/B12 deficiency. These data suggest that folate deficiency enhances the expression of smooth muscle marker gene expression, promotes a shift towards a skeletal muscle phenotype, and does not regulate gene expression via DNA methylation.",
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note = "Acknowledgements Microarray data shown in this manuscript have been deposited at NCBI via the Gene Expression Omnibus webpage under number: GSE125502. This work was funded by The Scottish Government Rural and Environment Science and Analytical Services Division (RESAS). The funder had no role in the design of the study, the analysis and interpretation of the data, or the publication process. The authors would like to thank Drs Bill Rees and Perry Barrett for helpful discussions throughout the project and on the manuscript. There are no competing interests to declare.",
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N1 - Acknowledgements Microarray data shown in this manuscript have been deposited at NCBI via the Gene Expression Omnibus webpage under number: GSE125502. This work was funded by The Scottish Government Rural and Environment Science and Analytical Services Division (RESAS). The funder had no role in the design of the study, the analysis and interpretation of the data, or the publication process. The authors would like to thank Drs Bill Rees and Perry Barrett for helpful discussions throughout the project and on the manuscript. There are no competing interests to declare.

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N2 - Elevated serum homocysteine, an intermediate of cellular one-carbon metabolism, is an independent risk factor for cardiovascular disease (CVD). Folate deficiency increases serum homocysteine and may contribute to CVD progression. Vascular smooth muscle cells regulate vascular contractility, but also contribute to repair processes in response to vascular injury. Nutritional deficiencies, like folate deficiency, are thought to impact on this phenotypic plasticity, possibly by epigenetic mechanisms. We have investigated the effect of folate deficiency on vascular smooth muscle cells in two cell culture systems representing early and late stages of smooth muscle cells differentiation. We find that folate deficiency promotes differentiation towards a more contractile phenotype as indicated by increased expression of respective marker genes. However, microarray analysis identified markers of striated muscle as the predominant gene expression change elicited by folate deficiency. These changes are not merely a reflection of cell cycle arrest, as foetal calf serum restriction or iron deficiency do not replicate the gene expression changes observed in response to folate deficiency. Folate deficiency only has a marginal effect on global DNA methylation. DNA methylation of CpG islands associated with genes regulated by folate deficiency remains unaffected. This supports our earlier findings in a mouse model system which also did not show any changes in global DNA methylation in response to folate and vitamin B6/B12 deficiency. These data suggest that folate deficiency enhances the expression of smooth muscle marker gene expression, promotes a shift towards a skeletal muscle phenotype, and does not regulate gene expression via DNA methylation.

AB - Elevated serum homocysteine, an intermediate of cellular one-carbon metabolism, is an independent risk factor for cardiovascular disease (CVD). Folate deficiency increases serum homocysteine and may contribute to CVD progression. Vascular smooth muscle cells regulate vascular contractility, but also contribute to repair processes in response to vascular injury. Nutritional deficiencies, like folate deficiency, are thought to impact on this phenotypic plasticity, possibly by epigenetic mechanisms. We have investigated the effect of folate deficiency on vascular smooth muscle cells in two cell culture systems representing early and late stages of smooth muscle cells differentiation. We find that folate deficiency promotes differentiation towards a more contractile phenotype as indicated by increased expression of respective marker genes. However, microarray analysis identified markers of striated muscle as the predominant gene expression change elicited by folate deficiency. These changes are not merely a reflection of cell cycle arrest, as foetal calf serum restriction or iron deficiency do not replicate the gene expression changes observed in response to folate deficiency. Folate deficiency only has a marginal effect on global DNA methylation. DNA methylation of CpG islands associated with genes regulated by folate deficiency remains unaffected. This supports our earlier findings in a mouse model system which also did not show any changes in global DNA methylation in response to folate and vitamin B6/B12 deficiency. These data suggest that folate deficiency enhances the expression of smooth muscle marker gene expression, promotes a shift towards a skeletal muscle phenotype, and does not regulate gene expression via DNA methylation.

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