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Upregulated miR-224-5p suppresses osteoblast differentiation by increasing the expression of Pai-1 in the lumbar spine of a rat model of congenital kyphoscoliosis


Congenital scoliosis is defined by the presence of structural anatomical malformations that arise from failures of vertebral formation or segmentation before and after birth. The understanding of genetic background and key genes for congenital scoliosis is still poor. We herein report that the excess expression of plasminogen activator inhibitor-1 (Pai-1) induced by the upregulation of miR-224-5p is involved in the pathogenesis of congenital kyphoscoliosis through impaired osteoblast differentiation. We first investigated the variety and progression of abnormalities of the lumbar spines in Ishibashi (IS) rats, a rat model of congenital kyphoscoliosis. The rats had already shown fusion and division of the primary ossification center at postnatal day 4. Over time, the rats showed various abnormalities of the lumbar spine, including the fusion of the annular epiphyseal nucleus. At postnatal day 42, spinal curvature was clearly observed due to the fusion of the vertebral bodies. Using a microRNA array, we found that the expression of miR-224-5p was increased in the lumbar spine of the rats at postnatal day 4. The expression of Pai-1, which is involved in osteoblast differentiation regulated by miR-224-5p, was also increased, while the levels of type I collagen, a marker of osteoblast differentiation, were decreased in the lumbar spine. These results indicate that the aberrant expression of miRNA-224-5p and its target genes is involved in the impaired osteoblast differentiation and may provide a partial molecular explanation for the pathogenesis of congenital scoliosis.

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This work was supported by a Grant-in-Aid for Scientific Research (C) (16K10809) from the JSPS (to H.I.) and the Mishima Kaiun Memorial Foundation (2015-3), Japan (to N.S.).

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SI, HS, and NS reproduced the congenital kyphoscoliosis model rats. SI, HS, DT, and NS conducted the experiments of gene and protein expression of miRNAs, Smads, Pai-1, and Col1A1. SI, HS, and YT performed the skeletal preparations and staining of rat lumber spines. SI, HI, HS, DT, HC, NK, and NS contributed to the supervision of the experiments and interpretation of the results. SI, HI, and NS designed the experiments and wrote the manuscript with input from all authors.

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Correspondence to Noriaki Shimokawa.

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Ishiwata, S., Iizuka, H., Sonoda, H. et al. Upregulated miR-224-5p suppresses osteoblast differentiation by increasing the expression of Pai-1 in the lumbar spine of a rat model of congenital kyphoscoliosis. Mol Cell Biochem 475, 53–62 (2020).

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  • Congenital scoliosis
  • Kyphoscoliosis model rats
  • microRNA array
  • miR-224-5p
  • Plasminogen activator inhibitor-1
  • Osteoblast differentiation