Asporin expression is highly regulated in human chondrocytes

TL;DR

This study investigates how asporin gene expression in human chondrocytes is regulated by cytokines, differentiation state, and the transcription factor Sp1, revealing complex control mechanisms relevant to osteoarthritis.

Contribution

It is the first to detail how cytokines and Sp1 regulate asporin expression in human chondrocytes, highlighting its role in cartilage biology.

Findings

Interleukin-1β and TNF-α downregulate ASPN

TGF-β1 upregulates ASPN in serum-free conditions

Sp1 increases ASPN expression via promoter interaction

Abstract

A significant association between a polymorphism in the D repeat of the gene encoding asporin and osteoarthritis, the most frequent of articular diseases, has been recently reported. The goal of the present study was to investigate the expression of this new class I small leucine-rich proteoglycan (SLRP) in human articular chondrocytes. First, we studied the modulation of asporin (ASPN) expression by cytokines by Western blot and reverse transcription-polymerase chain reaction. Interleukin-1$\beta$ and tumor necrosis factor-$\alpha$ downregulated ASPN, whereas transforming growth factor-$\beta$1 (when incubated in a serum-free medium) upregulated it. Similarly to proinflammatory cytokines, chondrocyte dedifferentiation induced by a successive passages of cells was accompanied by a decreased asporin expression, whereas their redifferentiation by three-dimensional culture restored its expression. Finally, we found an important role of the transcription factor Sp1 in the regulation of ASPN expression. Sp1 ectopic expression increased ASPN mRNA level and promoter activity. In addition, using gene reporter assay and electrophoretic mobility shift assay, we showed that Sp1 mediated its effect through a region located between -473 and -140 bp upstream of the transcription start site in ASPN gene. In conclusion, this report is the first study on the regulation of asporin expression by different cytokines in human articular chondrocytes. Our data indicate that the expression of this gene is finely regulated in cartilage and suggest a major role of Sp1.