Sclerostin is involved in osteogenic transdifferentiation of vascular smooth muscle cells in chronic kidney disease-associated vascular calcification with non-canonical Wnt signaling

Ren Fail. 2022 Dec;44(1):1426-1442. doi: 10.1080/0886022X.2022.2114370.

Abstract

Vascular calcification is prominent in patients with chronic kidney disease (CKD) and is a strong predictor of cardiovascular mortality in the CKD population. However, the mechanism underlying CKD-associated vascular calcification remains unclear. To identify potential therapeutic targets, a 5/6 nephrectomy rat model was established by feeding of a high-phosphorous diet as the CKD group and compared with sham group rats at 4 and 16 weeks. Sequencing analyses of the rat aorta revealed 643 upregulated and 1023 downregulated genes at 4 weeks, as well as 899 upregulated and 1185 downregulated genes at 16 weeks in the CKD group compared to the sham group. Bioinformatics analyses suggested that SOST (which encodes sclerostin) and Wnt signaling are involved in CKD-associated vascular calcification. Furthermore, protein-protein interactions analysis revealed interactions between SOST, WNT5A, and WNT5B, that involved runt-related transcription factor 2 (RUNX2) and transgelin (TAGLN). SOST was increased in CKD-associated vascular calcification following reduction of the Wnt signaling, including WNT5A and WNT5B, both in vivo and in vitro. TargetScan was used to predict the microRNAs (miRNAs) targeting WNT5A and WNT5B. The expression levels of miR-542-3p, miR-298-3p, miR-376b-5p, and miR-3568 were significantly reduced, whereas that of miR-742-3p was significantly increased in calcified rat aortic vascular smooth muscle cells (VSMCs). In CKD rat aortas, the expression of miR-542-3p, miR-298-3p, miR-376b-5p, miR-3568, miR-742-3p, and miR-22-5p were significantly reduced at both 4 and 16 weeks. Altogether, owing to several assessments, potentially diagnostic and prognostic biomarkers for improving common CKD diagnostic tools were identified in this study. Abbreviations: BUN: blood urea nitrogen; CKD: chronic kidney disease; CKD-MBD: chronic kidney disease-mineral bone disorder; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; GO: the Gene Ontology; HE: hematoxylin-eosin; HRP: horseradish peroxidase; KEGG: Kyoto Encyclopedia of Genes and Genomes; MiRNAs: microRNAs; PAS: periodic acid-Schiff; RUNX2: runt-related transcription factor 2; SCr: serum creatinine; STRING: the Search Tool for the Retrieval of Interacting Genes/Proteins; TAGLN: transgelin; VSMC: vascular smooth muscle cell.

Keywords: Chronic kidney disease; Wnt signaling pathway; microRNA; sclerostin; vascular calcification; vascular smooth muscle cells.

MeSH terms

  • Animals
  • Cell Transdifferentiation
  • Core Binding Factor Alpha 1 Subunit / genetics
  • Core Binding Factor Alpha 1 Subunit / metabolism
  • MicroRNAs* / genetics
  • MicroRNAs* / metabolism
  • Muscle, Smooth, Vascular / metabolism
  • Myocytes, Smooth Muscle / metabolism
  • Rats
  • Renal Insufficiency, Chronic* / metabolism
  • Vascular Calcification* / genetics
  • Vascular Calcification* / metabolism
  • Wnt Signaling Pathway

Substances

  • Core Binding Factor Alpha 1 Subunit
  • MIRN22 microRNA, rat
  • MicroRNAs

Grants and funding

This work was supported by the National Natural Science Foundation of China [U21A20349, 81700607, 8170742, 81800613, 81970641, and 82070690]; PHD Foundation of Sichuan Academy of Sciences & Sichuan Provincial People’s Hospital [2015BS05]; Fundamental Research Funds for the Central Universities from UESTC [ZYGX2019J105]; Key R&D projects in Sichuan Province [2019YFS0538, 2021YFS0372 and 2021YFS0370]; Department of Science and Technology of Sichuan Province [2020ZYD034]; Foundation of Health and Family Planning Commission in Sichuan [16PJ424]; Foundation of Sichuan Medical Association [S20071]; and Zhongguancun Nephrology & Blood Purification Innovation Alliance [NBPIA20QC0102].