Loss of methyl-CpG-binding domain protein 2 enhances endothelial angiogenesis and protects mice against hind-limb ischemic injury

Xiaoquan Rao, Jixin Zhong, Shu Zhang, Yushan Zhang, Qilin Yu, Ping Yang, Mong-Heng Wang, David J Fulton, Huidong Shi, Zheng Dong, Daowen Wang, Cong Yi Wang

Research output: Contribution to journalArticle

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Abstract

Background-: Despite intensive investigation, how DNA methylation influences endothelial function remains poorly understood. We used methyl-CpG-binding domain protein 2 (MBD2), an interpreter for DNA methylome-encoded information, to dissect the impact of DNA methylation on endothelial function in both physiological and pathophysiological states. Methods and results-: Human umbilical vein endothelial cells under normal conditions express moderate levels of MBD2, but knockdown of MBD2 by siRNA significantly enhanced angiogenesis and provided protection against H2O2-induced apoptosis. Remarkably, Mbd2 mice were protected against hind-limb ischemia evidenced by the significant improvement in perfusion recovery, along with increased capillary and arteriole formation. Loss of MBD2 activated endothelial survival and proangiogenic signals downstream of vascular endothelial growth factor signaling characterized by an increase in endothelial nitric oxide synthase (eNOS) and vascular endothelial growth factor receptor 2 expression, along with enhanced extracellular signal-regulated kinase 1/2 activation and BCL-2 expression. Mechanistic studies confirmed the methylation of CpG elements in the eNOS and vascular endothelial growth factor receptor 2 promoter. MBD2 binds to these methylated CpG elements and suppresses eNOS promoter activity. On ischemic insult, key endothelial genes such as eNOS and vascular endothelial growth factor receptor 2 undergo a DNA methylation turnover, and MBD2 interprets the changes of DNA methylation to suppress their expressions. Moreover, MBD2 modulation of eNOS expression is likely confined to endothelial cells because nonendothelial cells such as splenocytes fail to express eNOS after loss of MBD2. Conclusions-: We provided direct evidence supporting that DNA methylation regulates endothelial function, which forms the molecular basis for understanding how environmental insults (epigenetic factor) affect the genome to modify disease susceptibility. Because MBD2 itself does not affect the methylation of DNA and is dispensable for normal physiology in mice, it could be a viable epigenetic target for modulating endothelial function in disease states.

Original languageEnglish (US)
Pages (from-to)2964-2974
Number of pages11
JournalCirculation
Volume123
Issue number25
DOIs
StatePublished - Jun 28 2011

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Extremities
Nitric Oxide Synthase Type III
DNA Methylation
Wounds and Injuries
Vascular Endothelial Growth Factor Receptor-2
Epigenomics
MBD2 protein
Mitogen-Activated Protein Kinase 3
Mitogen-Activated Protein Kinase 1
Disease Susceptibility
Human Umbilical Vein Endothelial Cells
Arterioles
Methylation
Vascular Endothelial Growth Factor A
Small Interfering RNA
Ischemia
Endothelial Cells
Perfusion
Genome
Apoptosis

Keywords

  • DNA methylation
  • MBD2 protein
  • angiogenesis
  • endothelium
  • nitric oxide synthase type III

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine
  • Physiology (medical)

Cite this

Loss of methyl-CpG-binding domain protein 2 enhances endothelial angiogenesis and protects mice against hind-limb ischemic injury. / Rao, Xiaoquan; Zhong, Jixin; Zhang, Shu; Zhang, Yushan; Yu, Qilin; Yang, Ping; Wang, Mong-Heng; Fulton, David J; Shi, Huidong; Dong, Zheng; Wang, Daowen; Wang, Cong Yi.

In: Circulation, Vol. 123, No. 25, 28.06.2011, p. 2964-2974.

Research output: Contribution to journalArticle

Rao, Xiaoquan ; Zhong, Jixin ; Zhang, Shu ; Zhang, Yushan ; Yu, Qilin ; Yang, Ping ; Wang, Mong-Heng ; Fulton, David J ; Shi, Huidong ; Dong, Zheng ; Wang, Daowen ; Wang, Cong Yi. / Loss of methyl-CpG-binding domain protein 2 enhances endothelial angiogenesis and protects mice against hind-limb ischemic injury. In: Circulation. 2011 ; Vol. 123, No. 25. pp. 2964-2974.
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AU - Zhong, Jixin

AU - Zhang, Shu

AU - Zhang, Yushan

AU - Yu, Qilin

AU - Yang, Ping

AU - Wang, Mong-Heng

AU - Fulton, David J

AU - Shi, Huidong

AU - Dong, Zheng

AU - Wang, Daowen

AU - Wang, Cong Yi

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