Abstract
Purpose :
How long non-coding RNAs (lncRNAs) regulate angiogenesis, especially ocular angiogenesis, remains unclear. A novel human lncRNA, termed lncEGFL7OS, was recently identified by our lab. A series of assays revealed that lncEGFL7OS is required for human ocular angiogenesis at least partly through regulating its host genes EGFL7 and miR-126. The purpose of the project is to further study the functional mechanism of lncEGFL7OS in endothelial cells. We test the hypothesis that lncEGFL7OS regulates angiogenesis by regulating histone modification at the host gene locus.
Methods :
RNA interferences, microRNA mimic or adenovirus overexpression approaches were used to regulate lncEGFL7OS and miR126 expression in endothelial cells. Real-time PCR were employed to confirm the regulation of the host genes EGFL7 and miR-126 by lncEGFL7OS. Regulation of angiogenesis by lncEGFL7OS was examined using EC-fibroblast co-culture assays and ex vivo models. Chromatin remodeling of the host genes was analyzed by PCR after precipitation of chromatins with antibodies probing different histone modification status, including H3K4me1, H3K4me3 and H3K27ac3.
Results :
We found that knockdown of lncEGFL7OS represses angiogenesis in vitro and ex vivo by regulating the expression of its host genes EGFL7 and miR-126. Overexpression of lncEGFL7OS has the opposite effect, consistent with an increase in H3K27ac3 and H3K4me3 modification at the host gene locus.
Conclusions :
lncEGFL7OS regulates human ocular angiogenesis by regulating histone modification at its host gene locus. Manipulation lncRNA expression may have therapeutic implications for vascular retinopathies in humans.
This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.