Effect of miRNA26b knockout on vascular calcification [scRNA-seq]

Vascular calcification is the abnormal deposition of calcium phosphates within the blood vessels. Although it significantly contributes to the development of cardiovascular disease, much remains unknown about the mechanisms driving this process. Recent advances have shown that microRNAs (miRNAs) could be critical in regulating the biological processes driving vascular calcification. Here, we investigated whether miRNA-26b (miR-26b) could regulate vascular calcification. By performing scRNA-seq of the aorta of miR-26b knockout (miR-26bKO) mice, we identified cell-specific targets and demonstrated that components of the BMP signalling pathway are dysregulated in the aortic cell types. Overall design: To investigate the contribution of miRNA26b to the development of vascualr calcification in mice we generated miRNA26b knockout model (miRNA26bKO) . We performed scRNA-seq using the whole aorta from deliniate regualated pathways in individual cell types. The whole mouse aorta was harvested from 6-month-old wild-type and miR-26bKO mice (wildtype n = 3 and miR26bKO n = 3) to determine the transcriptomes. The aorta was dissected from surrounding tissue, and the perivascular fat was removed. Upon digestion of tissue using protocol as reported below, cells were isolated and sorted using a FACS Aria II instrument and BD FACSDiva software (BD Biosciences).