ABSTRACT
INTRODUCTION: Runx2 is one of the most studied transcription factors expressed in mesenchymal stem cells (MSCs) upon their commitment toward an osteogenic differentiation. During endochondral bone formation in vivo, Sox9 directly interacts with Runx2 and represses its activity; however, the role of Sox9 in direct osteogenesis in vitro has been largely overlooked. METHODS: Bone marrow-derived human MSCs (hMSCs) were cultured in vitro either in the control or osteogenic medium supplemented with dexamethasone (DEX). To further investigate the role of Sox9 in direct osteogenesis in vitro, hMSCs were treated with Sox9 siRNA. RESULTS: We show here that Sox9 is the key early indicator during in vitro osteogenic differentiation of hMSCs. Osteogenic induction leads to a significant decrease of Sox9 gene and protein expression by day 7. Treatment of hMSCs with Sox9 siRNA enhanced mineralization in vitro, suggesting that downregulation of Sox9 is involved in direct osteogenesis. siRNA knockdown of Sox9 did not in itself induce osteogenesis in the absence of DEX, indicating that other factors are still required. CONCLUSION: Screening of not preselected donors of different ages and gender (n=12) has shown that the Runx2/Sox9 ratio on day 7 is correlated to the (45)Ca incorporation on day 28. The impact of Sox9 downregulation in the mineralization of human MSCs in vitro indicates a so far unprecedented role of Sox9 as a major regulator of direct osteogenesis. We propose that the Runx2/Sox9 ratio is a promising, early, in vitro screening method for osteogenicity of human MSCs.
Subject(s)
Core Binding Factor Alpha 1 Subunit/metabolism , Mesenchymal Stem Cells/metabolism , Osteogenesis , SOX9 Transcription Factor/metabolism , Adult , Aged , Core Binding Factor Alpha 1 Subunit/genetics , Dexamethasone/pharmacology , Down-Regulation/drug effects , Electroporation , Female , Humans , Male , Mesenchymal Stem Cells/cytology , Mesenchymal Stem Cells/drug effects , Middle Aged , Osteogenesis/drug effects , Osteogenesis/genetics , RNA, Messenger/genetics , RNA, Messenger/metabolism , RNA, Small Interfering/metabolism , SOX9 Transcription Factor/genetics , Tissue Donors , Young AdultABSTRACT
Proper development of the vascular system as one of the earliest and most critical steps during vertebrate embryogenesis is ensured by the exact spatial and temporal control of gene expression in cells forming the vessel network. Whereas the regulation of vascular system development is well elucidated on the level of ligand-receptor signaling, the processes on the transcriptional level are much less understood. As the signaling mechanisms in embryogenesis and pathological conditions are similar, the study of embryonic blood vessel development is of great interest for the treatment of cardiovascular diseases and cancer. This review discusses two transcription factors, HOXA9 and VEZF1, which are relevant for endothelial biology but are excluded in the bulk of transcription factor references discussing endothelial biology. To our knowledge, there is no comprehensive overview of these two transcription factors available to date. Here, we summarize the current knowledge of human HOXA9 and VEZF1 biology and function, we detail their target genes and roles in endothelial biology and propose that HOXA9 and VEZF1 also deserve consideration as relevant transcriptional regulators of endothelial biology. Due to their broad role in multiple aspects of endothelial biology, they might potentially become interesting targets for therapeutic manipulation of pathological blood vessel growth.
