title: In vitro epigenetic reprogramming of human cardiac mesenchymal stromal cells into functionally competent cardiovascular precursors.
abstract: Adult human cardiac mesenchymal-like stromal cells (CStC) represent a relatively accessible cell type useful for therapy. In this light, their conversion into cardiovascular precursors represents a potential successful strategy for cardiac repair. The aim of the present work was to reprogram CStC into functionally competent cardiovascular precursors using epigenetically active small molecules. CStC were exposed to low serum (5% FBS) in the presence of 5 µM all-trans Retinoic Acid (ATRA), 5 µM Phenyl Butyrate (PB), and 200 µM diethylenetriamine/nitric oxide (DETA/NO), to create a novel epigenetically active cocktail (EpiC). Upon treatment the expression of markers typical of cardiac resident stem cells such as c-Kit and MDR-1 were up-regulated, together with the expression of a number of cardiovascular-associated genes including KDR, GATA6, Nkx2.5, GATA4, HCN4, NaV1.5, and α-MHC. In addition, profiling analysis revealed that a significant number of microRNA involved in cardiomyocyte biology and cell differentiation/proliferation, including miR 133a, 210 and 34a, were up-regulated. Remarkably, almost 45% of EpiC-treated cells exhibited a TTX-sensitive sodium current and, to a lower extent in a few cells, also the pacemaker I(f) current. Mechanistically, the exposure to EpiC treatment introduced global histone modifications, characterized by increased levels of H3K4Me3 and H4K16Ac, as well as reduced H4K20Me3 and H3s10P, a pattern compatible with reduced proliferation and chromatin relaxation. Consistently, ChIP experiments performed with H3K4me3 or H3s10P histone modifications revealed the presence of a specific EpiC-dependent pattern in c-Kit, MDR-1, and Nkx2.5 promoter regions, possibly contributing to their modified expression. Taken together, these data indicate that CStC may be epigenetically reprogrammed to acquire molecular and biological properties associated with competent cardiovascular precursors.
authors.affiliation: Dipartimento di Scienze Cliniche e di Comunità, Università di Milano, Milano, Italy.
authors.forename: Matteo
authors.initials: M
authors.lastname: Vecellio
authors.forename: Viviana
authors.initials: V
authors.lastname: Meraviglia
authors.forename: Simona
authors.initials: S
authors.lastname: Nanni
authors.forename: Andrea
authors.initials: A
authors.lastname: Barbuti
authors.forename: Angela
authors.initials: A
authors.lastname: Scavone
authors.forename: Dario
authors.initials: D
authors.lastname: DiFrancesco
authors.forename: Antonella
authors.initials: A
authors.lastname: Farsetti
authors.forename: Giulio
authors.initials: G
authors.lastname: Pompilio
authors.forename: Gualtiero I
authors.initials: GI
authors.lastname: Colombo
authors.forename: Maurizio C
authors.initials: MC
authors.lastname: Capogrossi
authors.forename: Carlo
authors.initials: C
authors.lastname: Gaetano
authors.forename: Alessandra
authors.initials: A
authors.lastname: Rossini
mesh_terms: D000328:Adult; D015415:Biomarkers / Q000378:metabolism; D015153:Blotting, Western; D002454:Cell Differentiation; D002843:Chromatin / Q000235:genetics; D047369:Chromatin Immunoprecipitation; D004594:Electrophysiology; D044127:Epigenesis, Genetic / Q000235:genetics*; D020869:Gene Expression Profiling; D006321:Heart / Q000502:physiology*; D006657:Histones / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D035683:MicroRNAs / Q000235:genetics; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D020030:Nitric Oxide Donors / Q000494:pharmacology; D020411:Oligonucleotide Array Sequence Analysis; D011401:Promoter Regions, Genetic / Q000235:genetics; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D014212:Tretinoin / Q000494:pharmacology; D015854:Up-Regulation
chemical_list: D015415:Biomarkers; D002843:Chromatin; D006657:Histones; D035683:MicroRNAs; D020030:Nitric Oxide Donors; D014212:Tretinoin