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A pituitary homeobox 2 (Pitx2):microRNA-200a-3p:β-catenin pathway converts mesenchymal cells to amelogenin-expressing dental epithelial cells
Journal article   Open access   Peer reviewed

A pituitary homeobox 2 (Pitx2):microRNA-200a-3p:β-catenin pathway converts mesenchymal cells to amelogenin-expressing dental epithelial cells

Thad Sharp, Jianbo Wang, Xiao Li, Huojun Cao, Shan Gao, Myriam Moreno and Brad A Amendt
The Journal of biological chemistry, Vol.289(39), pp.27327-27341
09/26/2014
DOI: 10.1074/jbc.M114.575654
PMCID: PMC4175363
PMID: 25122764
url
https://doi.org/10.1074/jbc.M114.575654View
Published (Version of record) Open Access

Abstract

Pitx2, Wnt/β-catenin signaling, and microRNAs (miRs) play a critical role in the regulation of dental stem cells during embryonic development. In this report, we have identified a Pitx2:β-catenin regulatory pathway involved in epithelial cell differentiation and conversion of mesenchymal cells to amelogenin expressing epithelial cells via miR-200a. Pitx2 and β-catenin are expressed in the labial incisor cervical loop or epithelial stem cell niche, with decreased expression in the differentiating ameloblast cells of the mouse lower incisor. Bioinformatics analyses reveal that miR-200a-3p expression is activated in the pre-ameloblast cells to enhance epithelial cell differentiation. We demonstrate that Pitx2 activates miR-200a-3p expression and miR-200a-3p reciprocally represses Pitx2 and β-catenin expression. Pitx2 and β-catenin interact to synergistically activate gene expression during odontogenesis and miR-200a-3p attenuates their expression and directs differentiation. To understand how this mechanism controls cell differentiation and cell fate, oral epithelial and odontoblast mesenchymal cells were reprogrammed by a two-step induction method using Pitx2 and miR-200a-3p. Conversion to amelogenin expressing dental epithelial cells involved an up-regulation of the stem cell marker Sox2 and proliferation genes and decreased expression of mesenchymal markers. E-cadherin expression was increased as well as ameloblast specific factors. The combination of Pitx2, a regulator of dental stem cells and miR-200a converts mesenchymal cells to a fully differentiated dental epithelial cell type. This pathway and reprogramming can be used to reprogram mesenchymal or oral epithelial cells to dental epithelial (ameloblast) cells, which can be used in tissue repair and regeneration studies.
Epithelial Cells - metabolism Homeodomain Proteins - metabolism Humans Mesenchymal Stromal Cells - metabolism Mice, Transgenic MicroRNAs - metabolism Transcription Factors - genetics beta Catenin - metabolism Homeodomain Proteins - genetics beta Catenin - genetics SOXB1 Transcription Factors - metabolism Transcription Factors - metabolism Incisor - cytology Mesenchymal Stromal Cells - cytology Animals Amelogenin - genetics SOXB1 Transcription Factors - genetics Incisor - metabolism Amelogenin - metabolism Mice MicroRNAs - genetics Epithelial Cells - cytology Cell Differentiation - physiology

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