Differential Promoter Methylation of Macrophage Genes Is Associated With Impaired Vascular Growth in Ischemic Muscles of Hyperlipidemic and Type 2 Diabetic XXXX : Genome-Wide Promoter Methylation Study.

@entity407 @entity1748 @entity73128 @entity29799 @entity23233 @entity6 @entity63 @entity19 @entity73 @entity21028 @entity9843 @entity73129 @entity59072 @entity10247
RATIONALE: @entity407 and @entity6 ( @entity6 ) severely impair adaptive vascular growth responses in ischemic muscles. This is largely attributed to dysregulated gene expression, although details of the changes are unknown. OBJECTIVE: To define the role of promoter methylation in adaptive vascular growth in @entity407 ( @entity1748 (-/-)ApoB(100/100)) and @entity6 ( @entity10247 / @entity1748 (-/-)ApoB(100/100)) @entity19 models of hindlimb @entity63 . METHODS AND RESULTS: Unilateral hindlimb @entity63 was induced by ligating femoral artery. Perfusion was assessed using ultrasound, and capillary and arteriole parameters were assessed using immunohistochemistry. Genome-wide methylated DNA sequencing was performed with DNA isolated from ischemic muscle, tissue macrophages (M  s), and endothelial cells. Compared with the controls, @entity407 and @entity6 @entity19 showed impaired perfusion recovery, which was associated with @entity73 and arteriogenesis. Genome-wide proximal promoter DNA methylation analysis suggested differential patterns of methylation in M   genes in ischemic muscles. Classically activated M1-M   gene promoters, including Cfb, @entity59072 , and @entity29799 , were significantly hypomethylated, whereas alternatively activated M2-M   gene promoters, including @entity21028 , @entity9843 , @entity73128 , @entity23233 , @entity73129 , and Fes, were significantly hypermethylated in M  s isolated from @entity407 and @entity6 ischemic muscles compared with controls. These results combined with mRNA expression and immunohistochemistry showed the predominance of proinflammatory M1-M  s, compared with anti-inflammatory and proangiogenic M2-M  s in @entity407 and @entity6 ischemic muscles. CONCLUSIONS: We found significant promoter hypomethylation of genes typical for proinflammatory M1-M  s and hypermethylation of anti-inflammatory, proangiogenic M2-M   genes in @entity407 and @entity6 ischemic muscles. Epigenetic alterations modify M   phenotype toward proinflammatory M1 as opposed to anti-inflammatory, proangiogenic, and tissue repair M2 phenotype, which may contribute to the impaired adaptive vascular growth under these pathological conditions.
