Nitric oxide determines mesodermic differentiation of mouse embryonic stem cells by activating class IIa histone deacetylases: potential therapeutic implications in a mouse model of hindlimb ischemia.
In human endothelial cells, nitric oxide (NO) results in class IIa histone deacetylases (HDACs) activation and marked histone deacetylation. It is unknown whether similar epigenetic events occur in embryonic stem cells (ESC) exposed to NO and how this treatment could influence ESC therapeutic potential during tissue regeneration.This study reports that the NO-dependent class IIa HDACs subcellular localization and activity decreases the global acetylation level of H3 histones in ESC and that this phenomenon is associated with the inhibition of Oct4, Nanog, and KLF4 expression. Further, a NO-induced formation of macromolecular complexes including HDAC3, 4, 7, and protein phosphatase 2A (PP2A) have been detected. These processes correlated with the expression of the mesodermal-specific protein brachyury (Bry) and the appearance of several vascular and skeletal muscle differentiation markers. These events were abolished by the class IIa-specific inhibitor MC1568 and by HDAC4 or HDAC7 short interfering RNA (siRNA). The ability of NO to induce mesodermic/cardiovascular gene expression prompted us to evaluate the regenerative potential of these cells in a mouse model of hindlimb ischemia. We found that NO-treated ESCs injected into the cardiac left ventricle selectively localized in the ischemic hindlimb and contributed to the regeneration of muscular and vascular structures. These findings establish a key role for NO and class IIa HDACs modulation in ESC mesodermal commitment and enhanced regenerative potential in vivo.
Spallotta
Francesco
F
Laboratorio di Patologia Vascolare, Istituto Dermopatico dell' Immacolata - Istituto di Ricovero e Cura a Carattere Scientifico, Rome, Italy.
Rosati
Jessica
J
Straino
Stefania
S
Nanni
Simona
S
Grasselli
Annalisa
A
Ambrosino
Valeria
V
Rotili
Dante
D
Valente
Sergio
S
Farsetti
Antonella
A
Mai
Antonello
A
Capogrossi
Maurizio C
MC
Gaetano
Carlo
C
Illi
Barbara
B
D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line; D049109:Cell Proliferation; D004195:Disease Models, Animal; D053595:Embryonic Stem Cells / Q000187:drug effects / Q000201:enzymology*; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects / Q000235:genetics; D006085:Graft Survival / Q000187:drug effects / Q000235:genetics; D056464:Histone Deacetylase 2 / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D006657:Histones / Q000187:drug effects / Q000378:metabolism; D007511:Ischemia / Q000628:therapy*; D000090062:Kruppel-Like Factor 4; D046911:Macromolecular Substances / Q000378:metabolism; D008297:Male; D008648:Mesoderm / Q000187:drug effects / Q000201:enzymology*; D051379:Mice; D008810:Mice, Inbred C57BL; D009569:Nitric Oxide / Q000378:metabolism* / Q000494:pharmacology; D020127:Recovery of Function / Q000187:drug effects / Q000235:genetics; D012038:Regeneration / Q000187:drug effects / Q000235:genetics; D033581:Stem Cell Transplantation / Q000379:methods
D015415:Biomarkers; D006657:Histones; C000715467:KLF4 protein, human; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D046911:Macromolecular Substances; D009569:Nitric Oxide; C533424:Hdac2 protein, mouse; D056464:Histone Deacetylase 2