METTL3-mediated <i>N</i><sup>6</sup>-methyladenosine modification governs pericyte dysfunction during diabetes-induced retinal vascular complication
Long Suo, Chang Liu, Qiuyang Zhang, Mu‐Di Yao, Yan Ma, Jin Yao, Qin Jiang, Biao Yan
Abstract
Rationale: Microvascular complication is a major cause of morbidity and mortality among the patients with diabetes. Pericyte dysfunction is the predominant pathological manifestation of microvascular complication. N 6 -methyladenosine (m 6 A) serves as the most prevalent modification in eukaryotic mRNAs. However, the role of m 6 A RNA modification in pericyte dysfunction is still unclear. Methods: Quantitative polymerase chain reactions and western blots were conducted to detect the change of m 6 A RNA modification in pericytes and mouse retinas following diabetic stress. MTT assay, transwell migration assay, caspase 3/7 activity assay, calcein-AM/propidium iodide (PI) staining, and TUNEL staining were conducted to determine the role of METTL3 in pericyte biology in vitro. Retinal trypsin digestion, vascular permeability assay, and IB4-NG2 double immunofluorescent staining were conducted to determine the role of METTL3 in retinal pericyte dysfunction and vascular complication. RNA sequencing, RNA pull-down assays and immunoblots were conducted to clarify the mechanism of METTL3-mediated pericyte dysfunction and vascular complication.