Research Article: MEPE-Derived ASARM Peptide Inhibits Odontogenic Differentiation of Dental Pulp Stem Cells and Impairs Mineralization in Tooth Models of X-Linked Hypophosphatemia

Date Published: February 22, 2013

Publisher: Public Library of Science

Author(s): Benjamin Salmon, Claire Bardet, Mayssam Khaddam, Jiar Naji, Benjamin R. Coyac, Brigitte Baroukh, Franck Letourneur, Julie Lesieur, Franck Decup, Dominique Le Denmat, Antonino Nicoletti, Anne Poliard, Peter S. Rowe, Eric Huet, Sibylle Opsahl Vital, Agnès Linglart, Marc D. McKee, Catherine Chaussain, Irina Kerkis. http://doi.org/10.1371/journal.pone.0056749

Abstract

Mutations in PHEX (phosphate-regulating gene with homologies to endopeptidases on the X-chromosome) cause X-linked familial hypophosphatemic rickets (XLH), a disorder having severe bone and tooth dentin mineralization defects. The absence of functional PHEX leads to abnormal accumulation of ASARM (acidic serine- and aspartate-rich motif) peptide − a substrate for PHEX and a strong inhibitor of mineralization − derived from MEPE (matrix extracellular phosphoglycoprotein) and other matrix proteins. MEPE-derived ASARM peptide accumulates in tooth dentin of XLH patients where it may impair dentinogenesis. Here, we investigated the effects of ASARM peptides in vitro and in vivo on odontoblast differentiation and matrix mineralization. Dental pulp stem cells from human exfoliated deciduous teeth (SHEDs) were seeded into a 3D collagen scaffold, and induced towards odontogenic differentiation. Cultures were treated with synthetic ASARM peptides (phosphorylated and nonphosphorylated) derived from the human MEPE sequence. Phosphorylated ASARM peptide inhibited SHED differentiation in vitro, with no mineralized nodule formation, decreased odontoblast marker expression, and upregulated MEPE expression. Phosphorylated ASARM peptide implanted in a rat molar pulp injury model impaired reparative dentin formation and mineralization, with increased MEPE immunohistochemical staining. In conclusion, using complementary models to study tooth dentin defects observed in XLH, we demonstrate that the MEPE-derived ASARM peptide inhibits both odontogenic differentiation and matrix mineralization, while increasing MEPE expression. These results contribute to a partial mechanistic explanation of XLH pathogenesis: direct inhibition of mineralization by ASARM peptide leads to the mineralization defects in XLH teeth. This process appears to be positively reinforced by the increased MEPE expression induced by ASARM. The MEPE-ASARM system can therefore be considered as a potential therapeutic target.

Partial Text

Mineralization is regulated by a number of determinants, including small molecules such as pyrophosphate, and proteins (including enzymes and hormones), that act either locally in the extracellular matrix and/or systemically to regulate calcium and phosphate metabolism. Currently, the complex links between these molecules are not well understood. Of note for tooth dentin (unlike bone) is that dentin is not involved in the regulation of the calcium and phosphate metabolism, and dentin is not remodeled. Thus, it is a tissue showing a permanent chronological record of disease that may be useful in deciphering these complex molecular interactions and mechanisms of mineralization.

A MEPE-derived ASARM peptide was previously identified in human hypophosphatemic tooth dentin [12]. Together with the results of the present study, these data present evidence that this peptide (and potentially other similar ASARM peptides derived from other matrix SIBLING proteins) may be involved in impairing tooth dentin mineralization as observed in patients affected with XLH. Using a cell culture collagen/tooth slice 3D scaffold model, together with an in vivo wound healing animal model, the present study demonstrates that the MEPE-derived ASARM peptide inhibits odontogenic differentiation of deciduous dental pulp stem cells (SHEDs), enhances MEPE expression, and impairs dentin ECM mineralization.

Source:

http://doi.org/10.1371/journal.pone.0056749

 

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