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EC4, a truncation of soluble N-cadherin, reduces vascular smooth muscle cell apoptosis and markers of atherosclerotic plaque instability.

Lyon CA, Johnson JL, White S, Sala-Newby GB, George SJ - Mol Ther Methods Clin Dev (2014)

Bottom Line: Interaction with FGFR in the extracellular (EC) 4 domain of SNC was essential for the antiapoptotic effect.EC4 significantly reduced smooth muscle cell, macrophage, and endothelial cell apoptosis in vitro by ~70%, similar to SNC.Elevation of plasma levels of EC4 in male apolipoprotein E-deficient mice with existing atherosclerosis significantly reduced apoptosis in brachiocephalic artery plaques by ~50%.

View Article: PubMed Central - PubMed

Affiliation: Bristol Heart Institute, Bristol Royal Infirmary , Bristol, UK.

ABSTRACT
Atherosclerotic plaque instability is precipitated by vascular smooth muscle cell apoptosis in the fibrous cap, weakening it and leading to plaque rupture. We previously showed that reducing smooth muscle cell apoptosis with soluble N-cadherin (SNC) increased features of plaque stability. We have now identified the active site of SNC and examined whether a truncated form containing this site retains the antiapoptotic effect. SNC was mutated to prevent interaction with N-cadherin or fibroblast growth factor receptor (FGFR). Interaction with FGFR in the extracellular (EC) 4 domain of SNC was essential for the antiapoptotic effect. Therefore, we made a truncated form consisting of the EC4 domain. EC4 significantly reduced smooth muscle cell, macrophage, and endothelial cell apoptosis in vitro by ~70%, similar to SNC. Elevation of plasma levels of EC4 in male apolipoprotein E-deficient mice with existing atherosclerosis significantly reduced apoptosis in brachiocephalic artery plaques by ~50%. EC4 reduced plaque size and the incidence of buried fibrous layers and the macrophage:smooth muscle cell ratio (surrogate markers of plaque instability). Interaction of EC4 with FGFR induced potent antiapoptotic signaling in vitro and in vivo. EC4 modulates atherosclerosis in mice demonstrating its therapeutic potential for retarding plaque size and instability.

No MeSH data available.


Related in: MedlinePlus

SNC-Fc and EC4 reduced apoptosis in macrophages and endothelial cells. Graphs show the percentage of apoptotic (a) macrophages and (b) endothelial cells as assessed by cleaved caspase-3 immunocytochemistry 24 hours after induction of apoptosis with Fas-L and treatment with the purified proteins (mean ± SEM, n = 3, *P < 0.05 versus Fc, fibroblast growth factor receptor (FGFR) mutated and EC4 mutant). EC, extracellular; HUVECs, human umbilical vein endothelial cells; SNC, soluble N-cadherin.
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fig2: SNC-Fc and EC4 reduced apoptosis in macrophages and endothelial cells. Graphs show the percentage of apoptotic (a) macrophages and (b) endothelial cells as assessed by cleaved caspase-3 immunocytochemistry 24 hours after induction of apoptosis with Fas-L and treatment with the purified proteins (mean ± SEM, n = 3, *P < 0.05 versus Fc, fibroblast growth factor receptor (FGFR) mutated and EC4 mutant). EC, extracellular; HUVECs, human umbilical vein endothelial cells; SNC, soluble N-cadherin.

Mentions: Macrophage and endothelial cell apoptosis are also observed during atherosclerotic plaque progression. EC4-Fc reduced apoptosis of blood monocyte–derived macrophages (Figure 2a) and human umbilical vein endothelial cells (Figure 2b) to an extent similar to that of SNC-Fc. As with VSMCs, this effect was ablated when the FGFR binding site was mutated (Figure 2a,b).


EC4, a truncation of soluble N-cadherin, reduces vascular smooth muscle cell apoptosis and markers of atherosclerotic plaque instability.

Lyon CA, Johnson JL, White S, Sala-Newby GB, George SJ - Mol Ther Methods Clin Dev (2014)

SNC-Fc and EC4 reduced apoptosis in macrophages and endothelial cells. Graphs show the percentage of apoptotic (a) macrophages and (b) endothelial cells as assessed by cleaved caspase-3 immunocytochemistry 24 hours after induction of apoptosis with Fas-L and treatment with the purified proteins (mean ± SEM, n = 3, *P < 0.05 versus Fc, fibroblast growth factor receptor (FGFR) mutated and EC4 mutant). EC, extracellular; HUVECs, human umbilical vein endothelial cells; SNC, soluble N-cadherin.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4362368&req=5

fig2: SNC-Fc and EC4 reduced apoptosis in macrophages and endothelial cells. Graphs show the percentage of apoptotic (a) macrophages and (b) endothelial cells as assessed by cleaved caspase-3 immunocytochemistry 24 hours after induction of apoptosis with Fas-L and treatment with the purified proteins (mean ± SEM, n = 3, *P < 0.05 versus Fc, fibroblast growth factor receptor (FGFR) mutated and EC4 mutant). EC, extracellular; HUVECs, human umbilical vein endothelial cells; SNC, soluble N-cadherin.
Mentions: Macrophage and endothelial cell apoptosis are also observed during atherosclerotic plaque progression. EC4-Fc reduced apoptosis of blood monocyte–derived macrophages (Figure 2a) and human umbilical vein endothelial cells (Figure 2b) to an extent similar to that of SNC-Fc. As with VSMCs, this effect was ablated when the FGFR binding site was mutated (Figure 2a,b).

Bottom Line: Interaction with FGFR in the extracellular (EC) 4 domain of SNC was essential for the antiapoptotic effect.EC4 significantly reduced smooth muscle cell, macrophage, and endothelial cell apoptosis in vitro by ~70%, similar to SNC.Elevation of plasma levels of EC4 in male apolipoprotein E-deficient mice with existing atherosclerosis significantly reduced apoptosis in brachiocephalic artery plaques by ~50%.

View Article: PubMed Central - PubMed

Affiliation: Bristol Heart Institute, Bristol Royal Infirmary , Bristol, UK.

ABSTRACT
Atherosclerotic plaque instability is precipitated by vascular smooth muscle cell apoptosis in the fibrous cap, weakening it and leading to plaque rupture. We previously showed that reducing smooth muscle cell apoptosis with soluble N-cadherin (SNC) increased features of plaque stability. We have now identified the active site of SNC and examined whether a truncated form containing this site retains the antiapoptotic effect. SNC was mutated to prevent interaction with N-cadherin or fibroblast growth factor receptor (FGFR). Interaction with FGFR in the extracellular (EC) 4 domain of SNC was essential for the antiapoptotic effect. Therefore, we made a truncated form consisting of the EC4 domain. EC4 significantly reduced smooth muscle cell, macrophage, and endothelial cell apoptosis in vitro by ~70%, similar to SNC. Elevation of plasma levels of EC4 in male apolipoprotein E-deficient mice with existing atherosclerosis significantly reduced apoptosis in brachiocephalic artery plaques by ~50%. EC4 reduced plaque size and the incidence of buried fibrous layers and the macrophage:smooth muscle cell ratio (surrogate markers of plaque instability). Interaction of EC4 with FGFR induced potent antiapoptotic signaling in vitro and in vivo. EC4 modulates atherosclerosis in mice demonstrating its therapeutic potential for retarding plaque size and instability.

No MeSH data available.


Related in: MedlinePlus