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

Abstract

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 mesodermalspecific 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. STEM CELLS 2010;28:431-442 Disclosure of potential conflicts of interest is found at the end of this article.

NO/beta-catenin crosstalk modulates primitive streak formation prior to embryonic stem cell osteogenic differentiation

“…While NO has been reported to control T-Bra levels in ESCs recently (Spallotta et al, 2010), we show here for the first time that NO potentially crosstalks with Wnt/CatnB signaling during primitive streak formation in ESCs to regulate T-Bra. Previous studies have speculated that this NO/Wnt crosstalk plays a role in colon cancer, leukemia, and colitis (Williams et al, 2003;Nath et al, 2005;Wang et al, 2009).…”

Splicing of Histone Deacetylase 7 Modulates Smooth Muscle Cell Proliferation and Neointima Formation Through Nuclear β-Catenin Translocation

“…6 HDAC7 is a member of the class II HDACs and has been linked to cardiovascular diseases. [7][8][9][10][11][12][13] Several pioneering studies have demonstrated that HDAC7 plays a crucial role in early embryonic development, 7 angiogenesis, 8,9 and SMC differentiation. 11 Moreover, HDAC7 can suppress endothelial cell proliferation 12 and responds to growth factors such as vascular endothelial growth factor and PDGF.…”

Pharmacologic Inhibition of Cardiac Stem Cell Senescence

“…Although NO production and endothelial nitric oxide synthase have been shown to be greater in longer living rodents, NO donors do not seem to influence animal maximum lifespan (Csiszar, et al, 2007). However, it has been shown both that NO can regulate telomerase activity (Farsetti, et al, 2009) and that it has a profound impact on mouse embryonic stem cell differentiation towards a cardiovascular fate (Spallotta, et al, 2010). Data on the effects of NO on hCSC are still missing.…”