Endothelial NOS is required for SDF-1α/CXCR4-mediated peripheral endothelial adhesion of c-kit+ bone marrow stem cells

Abstract

In the era of intravascular approaches for regenerative cell therapy, the underlying mechanisms of stem cell migration to non-marrow tissue have not been clarified. We hypothesized that next to a local inflammatory response implying adhesion molecule expression, endothelial nitric oxide synthase (eNOS)-dependent signaling is required for stromalcell-derived factor-1 alpha (SDF-1a)-induced adhesion of c-kit þ cells to the vascular endothelium. SDF-1a/tumor necrosis factor-alpha (TNF-a)-induced c-kit þ -cell shape change and migration capacity was studied in vitro using immunohistochemistry and Boyden chamber assays. In vivo interaction of c-kit þ cells from bone marrow with the endothelium in response to SDF-1a/TNF-a stimulation was visualized in the cremaster muscle microcirculation of wild-type (WT) and eNOS (À/À) mice using intravital fluorescence microscopy. In addition, NOS activity was inhibited with N-nitro-Larginine-methylester-hydrochloride in WT mice. To reveal c-kit þ -specific adhesion behavior, endogenous leukocytes (EL) and c-kit þ cells from peripheral blood served as control. Moreover, intercellular adhesion molecule-1 (ICAM-1) and CXCR4 were blocked systemically to determine their role in inflammation-related c-kit þ -cell adhesion. In vitro, SDF-1a enhanced c-kit þ -cell migration. In vivo, SDF-1a alone triggered endothelial rolling-not firm adherence-of c-kit þ cells in WT mice. While TNF-a alone had little effect on adhesion of c-kit þ cells, it induced maximum endothelial EL adherence. However, after combined treatment with SDF-1a þ TNF-a, endothelial adhesion of c-kit þ cells increased independent of their origin, while EL adhesion was not further incremented. Systemic treatment with anti-ICAM-1 and anti-CXCR4-monoclonal antibody completely abolished endothelial c-kit þ -cell adhesion. In N-nitro-L-arginine-methylester-hydrochloride-treated WT mice as well as in eNOS (À/À) mice, firm endothelial adhesion of c-kit þ cells was entirely abrogated, while EL adhesion was significantly increased. The chemokine SDF-1a mediates firm adhesion c-kit þ cells only in the presence of TNF-a stimulation via an ICAM-1-and CXCR4-dependent mechanism. The presence of eNOS appears to be a crucial and specific factor for firm c-kit þ -cell adhesion to the vascular endothelium.

SDF-1α stimulates JNK3 activity via eNOS-dependent nitrosylation of MKP7 to enhance endothelial migration

“…SDF-1␣ activates Akt/eNOS and MAP kinases including ERK1/2 and p38 in different cell types (including endothelial cells and EPCs), resulting in a vast array of consequences including cell migration, apoptosis, and cell survival (27)(28)(29)(30). However, until now, no detailed description of the signaling pathways involved in SDF-1␣-dependent angiogenesis have been published.…”

Long term effects of the implantation of autologous bone marrow mononuclear cells for type 2 diabetes mellitus

“…The number of EPCs in patients with T2DM is reduced and the risk of cardiovascular disease is increased. This is as a result of multiple factors [3,27,[37][38][39]. EPCs can augment vascular repair and revascularization [40].…”

Impact of Reactive Oxygen and Reactive Nitrogen Species for Stem Cell Mobilization, Function and Cardiovascular Differentiation

“…eNOS in the host myocardium promoted mesenchymal stem cell migration to the ischemic myocardium and improved cardiac function through cGMP-dependent increases in SDF-1α expression ). The local inflammatory response implying adhesion molecule expression and eNOS-dependent signaling was required for SDF-1α-induced adhesion of c-kit + cells to the vascular endothelium (Kaminski et al, 2008). Furthermore, oxidative stress from lactate metabolism by circulating stem/progenitor cells accelerated further stem cell recruitment and differentiation through thioredoxin-1 (Trx1)-mediated elevations in HIF-1 levels and the subsequent synthesis of HIF-1-dependent growth factors including VEGF and SDF-1α (Milovanova et al, 2008).…”