Chemical and genetic blockade of HDACs enhances osteogenic differentiation of human adipose tissue-derived stem cells by oppositely affecting osteogenic and adipogenic transcription factors
Authors: Stefania Niada
Paola Bendinelli
Anna Teresa Brini
Maria Alfonsina Desiderio
Laura de Girolamo
Emanuela Matteucci
Elena Arrigoni
Paola Maroni
Journal: Biochemical and Biophysical Research Communications (2012)
Discipline: BIOLOGY & BIOCHEMISTRY
Keywords: hASCs; acetylation; Runt-related transcription factor 2 (Runx2); Peroxisome proliferator activated receptor (PPAR) γ; osteogenic
Affiliations: Istituto Ortopedico Galeazzi, Milano, Italy.

Abstract:  The human adipose-tissue derived stem/stromal cells (hASCs) are an interesting source for bone-tissue engineering applications. Our aim was to clarify in hASCs the role of acetylation in the control of Runt-related transcription factor 2 (Runx2) and Peroxisome proliferator activated receptor (PPAR) γ. These key osteogenic and adipogenic transcription factors are oppositely involved in osteo-differentiation. The hASCs, committed or not towards bone lineage with osteoinductive medium, were exposed to HDACs chemical blockade with Trichostatin A (TSA) or were genetically silenced for HDACs. Alkaline phosphatase (ALP) and collagen/calcium deposition, considered as early and late osteogenic markers, were evaluated concomitantly as index of osteo-differentiation. TSA pretreatment, useful experimental protocol to analyse pan-HDAC-chemical inhibition, and switch to osteogenic medium induced early-osteoblast maturation gene Runx2, while transiently decreased PPARγ and scarcely affected late-differentiation markers. Time-dependent effects were observed after knocking-down of HDAC1 and 3: Runx2 and ALP underwent early activation, followed by late-osteogenic markers increase and by PPARγ/ALP activity diminutions mostly after HDAC3 silencing. HDAC1 and 3 genetic blockade increased and decreased Runx2 and PPARγ target genes, respectively. Noteworthy, HDACs knocking-down favoured the commitment effect of osteogenic medium. Our results reveal a role for HDACs in orchestrating osteo-differentiation of hASCs at transcriptional level, and might provide new insights into the modulation of hASCs-based regenerative therapy. 

Retracted: No

PubPeer comments:
 - Figure 4C BBA 1813 and Figure 5 Mol Cancer Res 7, 1328 For HDAC1 and HDAC3 we reported our results in the two papers (Mol Cancer Res 7 and BBA 1813). So, also the vinculins are repeated - The other vinculins in the yellow and blue squares are not the same bands undergoing vertical or horizontal flips (Figure 5 Carcinogenesis 30, Figure 1B BBA 1843, Figure 2 European J Cancer 49). -HDAC1, Figure 5 Mol Cancer Res: when you compare the original pictures in the two articles, it is possible to appreciate that the 3 evidenced (yellow) bands are not the same of those in Figure 5 Carcinogenesis. In fact, in the original pictures they show different thickness. HDAC3, Figure 5 Mol Cancer Res: the vinculins shows some differences from those of Figure 2A BBRC. By simple observation, in Figure5 Carcinogenesis 30, the two bands indicated (red) do not seem stretched; the flip of the bands in blue and green squares is difficult to discern (Carcinogenesis 30 vs. Figure 1B BBA 1843 and Figure 1B Exp Cell Res 319). , 
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