ALKBH1-demethylated DNA N6-methyladenine modification triggers vascular calcification via osteogenic reprogramming in chronic kidney disease.

Ouyang, Liu; Su, Xiaoyan; Li, Wenxin; Tang, Liangqiu; Zhang, Mengbi; Zhu, Yongjun; Xie, Changming; Zhang, Puhua; Chen, Jie; Huang, Hui

Vascular calcification (VC) predicts cardiovascular morbidity and mortality in chronic kidney disease (CKD). To date, the underlying mechanisms remain unclear. We detected leukocyte DNA N6-methyladenine (6mA) levels in CKD patients with or without aortic arch calcification. We used arteries from CKD mice infected with vascular smooth muscle cells (VSMCs)-targeted adeno-associated virus encoding alkB homolog 1 (Alkbh1) gene or Alkbh1 shRNA to evaluate features of calcification. We identified that leukocyte 6mA levels were significantly reduced as the severity of VC increased in CKD patients. Decreased 6mA demethylation resulted from the upregulation of ALKBH1. Here, ALKBH1 overexpression aggravated, whereas its depletion blunted VC progression and osteogenic reprogramming in vivo and in vitro. Mechanistically, ALKBH1-demethylated DNA 6mA modification could facilitate the binding of octamer-binding transcription factor 4 (Oct4) to bone morphogenetic protein 2 (BMP2) promoter and activate BMP2 transcription. This resulted in osteogenic reprogramming of VSMCs and subsequent VC progression. Either BMP2 or Oct4 depletion alleviated the pro-calcifying effects of ALKBH1. This suggests targeting ALKBH1 might be a therapeutic method to reduce the burden of VC in CKD.