Orphan Nuclear Receptor NR4A3 Promotes Vascular Calcification via Histone Lactylation

Wenqi Ma; Kangni Jia; Haomai Cheng; Hong Xu; Zhigang Li; Hang Zhang; Hongyang Xie; Lingfang Zhuang; Ziyang Wang; Yuke Cui; Hang Sun; Lei Yi; Zhiyong Chen; Shengzhong Duan; Motoaki Sano; Keiichi Fukuda; Lin Lu; Fei Gao; Ruiyan Zhang; Xiaoxiang Yan

doi:10.1161/CIRCRESAHA.123.323699

Orphan Nuclear Receptor NR4A3 Promotes Vascular Calcification via Histone Lactylation

Circ Res. 2024 Apr 17. doi: 10.1161/CIRCRESAHA.123.323699. Online ahead of print.

Authors

Wenqi Ma^#^{1

2}, Kangni Jia^#^{1

2}, Haomai Cheng^#^{1

2}, Hong Xu³, Zhigang Li^{1

2}, Hang Zhang^{1

2}, Hongyang Xie^{1

2}, Lingfang Zhuang^{1

2}, Ziyang Wang^{1

2}, Yuke Cui^{1

2}, Hang Sun^{1

2}, Lei Yi^{1

2}, Zhiyong Chen^{1

2}, Shengzhong Duan^{4

5

6}, Motoaki Sano⁷, Keiichi Fukuda⁷, Lin Lu^{1

2}, Fei Gao⁸, Ruiyan Zhang^{1

2}, Xiaoxiang Yan^{1

2}

Affiliations

¹ Department of Cardiovascular Medicine, Ruijin Hospital (W.M., K.J., H.C., Z.L., H.Z., H.X., L.Z., Z.W., Y.C., H.S., L.Y., Z.C., L.L., R.Z., X.Y.).
² Institute of Cardiovascular Diseases (W.M., K.J., H.C., Z.L., H.Z., H.X., L.Z., Z.W., Y.C., H.S., L.Y., Z.C., L.L., R.Z., X.Y.).
³ Department of Cardiovascular Surgery, Ruijin Hospital (H.X.).
⁴ Laboratory of Oral Microbiota and Systemic Diseases, Shanghai Ninth People's Hospital, College of Stomatology (S.D.).
⁵ Department of Cardiovascular Medicine, State Key Laboratory of Medical Genomics, Shanghai Key Laboratory of Hypertension, Shanghai Institute of Hypertension, Ruijin Hospital (S.D.).
⁶ Shanghai Jiao Tong University School of Medicine, China. National Center for Stomatology, National Clinical Research Center for Oral Diseases, Shanghai Key Laboratory of Stomatology, China (S.D.).
⁷ Department of Cardiology, Keio University School of Medicine, Tokyo, Japan (M.S., K.F.).
⁸ Beijing Anzhen Hospital, Capital Medical University, China (F.G.).

^# Contributed equally.

PMID: 38629274
DOI: 10.1161/CIRCRESAHA.123.323699

Abstract

Background: Medial arterial calcification is a chronic systemic vascular disorder distinct from atherosclerosis and is commonly observed in patients with chronic kidney disease, diabetes, and aging individuals. We previously showed that NR4A3 (nuclear receptor subfamily 4 group A member 3), an orphan nuclear receptor, is a key regulator in apo (apolipoprotein) A-IV-induced atherosclerosis progression; however, its role in vascular calcification is poorly understood.

Methods: We generated NR4A3^-/- mice and 2 different types of medial arterial calcification models to investigate the biological roles of NR4A3 in vascular calcification. RNA-seq was performed to determine the transcriptional profile of NR4A3^-/- vascular smooth muscle cells under β-glycerophosphate treatment. We integrated CUT&Tag analysis and RNA-seq data to further investigate the gene regulatory mechanisms of NR4A3 in arterial calcification and target genes regulated by histone lactylation.

Results: NR4A3 expression was upregulated in calcified aortic tissues from chronic kidney disease mice, 1,25(OH)₂VitD₃ overload-induced mice, and human calcified aorta. NR4A3 deficiency preserved the vascular smooth muscle cell contractile phenotype, inhibited osteoblast differentiation-related gene expression, and reduced calcium deposition in the vasculature. Further, NR4A3 deficiency lowered the glycolytic rate and lactate production during the calcification process and decreased histone lactylation. Mechanistic studies further showed that NR4A3 enhanced glycolysis activity by directly binding to the promoter regions of the 2 glycolysis genes ALDOA and PFKL and driving their transcriptional initiation. Furthermore, histone lactylation promoted medial calcification both in vivo and in vitro. NR4A3 deficiency inhibited the transcription activation and expression of Phospho1 (phosphatase orphan 1). Consistently, pharmacological inhibition of Phospho1-attenuated calcium deposition in NR4A3-overexpressed vascular smooth muscle cells, whereas overexpression of Phospho1 reversed the anticalcific effect of NR4A3 deficiency in vascular smooth muscle cells.

Conclusions: Taken together, our findings reveal that NR4A3-mediated histone lactylation is a novel metabolome-epigenome signaling cascade mechanism that participates in the pathogenesis of medial arterial calcification.

Keywords: glycolysis; histone lactylation; phospho1; vascular calcification; vascular smooth muscle cell.