Transcriptomic profiling of endothelial cells with altered cholesterol

Cholesterol has been associated with the regulation of angiogenesis in endothelial cells of the central nervous system, but the mechanisms responsible for this regulation have not been explored at the genome level. Here, we performed transcriptomic profiling on the isolated brain vasculature of mouse embryos lacking two main cholesterol transporters, ABCA1 and SR-B1. We complemented these data with transcriptomic profiles of a cell line representing brain endothelial cells and chicken chorioallantoic membranes treated with methyl--cyclodextrin, which removes cholesterol from cells. We observed changes in the expression of many angiogenic genes upon modulation of cellular cholesterol, indicating coupling of cholesterol metabolism and angiogenesis in CNS endothelial cells. We obtained brain vascular fragments from fetuses lacking ABCA1 or SR-B1 and their respective controls at E18.5. We also incubated bEnd.3 cells and chicken chorioallantoic membranes with methyl--cyclodextrin in vitro and in ovo. PoliA+ libraries were constructed from extracted total RNA, and sequenced in PE150 mode to produce around 30 million reads per sample.