Frontiers in cell and developmental biology
Methylglyoxal hampers the angiogenic capacity of human retinal endothelial cells modifying cell transcriptome with the contribution of NRSF/REST
By Aprile M, Nigro C, Perfetto C and 9 others.
Abstract
Background
The functional impairment of retinal microvascular cells is an early and central event in diabetic retinopathy (DR). Methylglyoxal (MGO), the most reactive precursor of advanced glycation end products (AGEs), has been implicated in vascular injury and neuro-retinal dysfunction. Its direct effects on retinal endothelial cells and underlying molecular mechanisms remain poorly understood.
Methods
Human retinal endothelial cells (hRECs) were exposed to MGO and the effect of sublethal doses on the proangiogenic function was evaluated by migration, tube formation and pericyte recruitment assays. MGO derived AGEs were evaluated by western blot. Transcriptome and miRNome profiling were performed to identify gene expression and microRNA perturbations. Integrative in silico analyses combining RNA-Seq with ENCODE ChIP-Seq datasets were used to identify transcriptional regulators responsible for MGO-induced gene deregulation. Transcriptomic profiles were compared with independent human DR datasets, and REST was silenced by siRNAs followed by analyses of proangiogenic function.