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ALKBH1-demethylated DNA N6-methyladenine modification triggers vascular calcification via osteogenic reprogramming in

Liu Ouyang1,2, Xiaoyan Su3, Wenxin Li2

  • 1Department of Cardiology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China.

The Journal of Clinical Investigation
|May 18, 2021
PubMed

Insights

In chronic kidney disease (CKD), reduced leukocyte DNA N6-methyladenine (6mA) correlates with vascular calcification (VC). ALKBH1 upregulation drives VC by promoting osteogenic reprogramming via Oct4 and BMP2.

Area of Science:

  • Molecular Biology
  • Nephrology
  • Cardiovascular Research

Background:

  • Vascular calcification (VC) is a significant predictor of cardiovascular morbidity and mortality in chronic kidney disease (CKD).
  • The precise molecular mechanisms underlying VC in CKD remain incompletely understood.
  • Leukocyte DNA N6-methyladenine (6mA) levels have not been previously investigated in relation to VC severity in CKD patients.

Purpose of the Study:

  • To investigate the role of leukocyte DNA N6-methyladenine (6mA) modifications in vascular calcification (VC) among patients with chronic kidney disease (CKD).
  • To elucidate the mechanistic link between ALKBH1, DNA 6mA demethylation, and the osteogenic reprogramming of vascular smooth muscle cells (VSMCs) in VC.
  • To evaluate ALKBH1 as a potential therapeutic target for mitigating VC in CKD.

Main Methods:

  • Detection of leukocyte DNA N6-methyladenine (6mA) levels in CKD patients with varying degrees of aortic arch calcification.
  • Utilizing vascular smooth muscle cell-targeted (VSMC-targeted) adeno-associated virus (AAV) vectors encoding Alkbh1 gene or Alkbh1 shRNA in CKD mouse models to assess VC.
  • In vitro and in vivo experiments to evaluate the impact of ALKBH1 overexpression or depletion on VC progression and VSMC osteogenic reprogramming, including assessment of BMP2 and Oct4 involvement.

Main Results:

  • Leukocyte 6mA levels were significantly decreased with increasing severity of VC in CKD patients.
  • Upregulation of ALKBH1 was associated with decreased 6mA demethylation.
  • ALKBH1 overexpression exacerbated VC and osteogenic reprogramming, while its depletion attenuated these processes in vitro and in vivo.
  • ALKBH1-mediated DNA 6mA demethylation facilitated Oct4 binding to the BMP2 promoter, activating BMP2 transcription and promoting VSMC osteogenic reprogramming.
  • Depletion of either BMP2 or Oct4 mitigated the pro-calcifying effects of ALKBH1.

Conclusions:

  • Reduced leukocyte DNA 6mA is a potential biomarker for vascular calcification severity in CKD.
  • ALKBH1 plays a critical role in promoting VC in CKD by driving VSMC osteogenic reprogramming through the Oct4/BMP2 pathway.
  • Targeting ALKBH1 presents a promising therapeutic strategy for reducing the burden of vascular calcification in CKD patients.

Keywords:
CardiologyCardiovascular diseaseChronic kidney diseaseEpigeneticsVascular Biology

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