title: Chemical and genetic blockade of HDACs enhances osteogenic differentiation of human adipose tissue-derived stem cells by oppositely affecting osteogenic and adipogenic transcription factors.
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.
authors.affiliation: Istituto Ortopedico Galeazzi, Milano, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Maroni
authors.forename: Anna Teresa
authors.initials: AT
authors.lastname: Brini
authors.forename: Elena
authors.initials: E
authors.lastname: Arrigoni
authors.forename: Laura
authors.initials: L
authors.lastname: de Girolamo
authors.forename: Stefania
authors.initials: S
authors.lastname: Niada
authors.forename: Emanuela
authors.initials: E
authors.lastname: Matteucci
authors.forename: Paola
authors.initials: P
authors.lastname: Bendinelli
authors.forename: Maria Alfonsina
authors.initials: MA
authors.lastname: Desiderio
mesh_terms: D050156:Adipogenesis / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D000273:Adipose Tissue / Q000166:cytology* / Q000187:drug effects; D000328:Adult; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D002478:Cells, Cultured; D050659:Core Binding Factor Alpha 1 Subunit / Q000378:metabolism; D005260:Female; D055785:Gene Knockdown Techniques; D056284:Histone Deacetylase 1 / Q000235:genetics; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology; D006655:Histone Deacetylases / Q000235:genetics / Q000502:physiology*; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D008875:Middle Aged; D010012:Osteogenesis / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D047495:PPAR gamma / Q000378:metabolism; D044968:Regenerative Medicine; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects; D013489:Suppression, Genetic
chemical_list: D050659:Core Binding Factor Alpha 1 Subunit; D056572:Histone Deacetylase Inhibitors; D006877:Hydroxamic Acids; D047495:PPAR gamma; C493709:RUNX2 protein, human; C012589:trichostatin A; C110550:HDAC1 protein, human; D056284:Histone Deacetylase 1; D006655:Histone Deacetylases