Comprehensive Circular RNA Profiling Of Proliferative Vitreoretinopathy

Objective:Proliferative vitreoretinopathy?PVR?is a complication that can follow rhegmatogenous retinal detachments and penetrating injury,as well as intraocular surgery.Circular RNAs?circRNAs?are a novel type of RNA transcripts,and they have emerged as important regulators in many biological processes and disease development.However,characteristics and functions of circRNAs in PVR remains elusive.This study aims to investigate the role of circRNAs in PVR and its clinical significance.Methods:1.Microarray analysis was conducted to detect PVR-related circRNAs.Quantitative reverse transcription polymerase chain reactions?qRT-PCRs?were used randomly to verify the expression pattern of some of these differentially expressed circRNAs,and we selected the most up-regulated circRNA as the target for the following study.2.qRT-PCRs was used to compare the target circRNA expression in retinas of trauma patients,PVR-ERMs,and secondary ERMs developed after cataract surgery.Then,we detected the expression of PVR markers PDGFA,PDGFC and KNG1 in traumatic retinas,PVR-ERMs,and secondary ERMs.In addition,qRT-PCRs were used to detect the target circRNA expression in serum and vitreous of different PVR patients.3.The expression of target circRNA in Retinal Pigment Epithelial cells?RPE?was down-regulated or up-regulated by cell transfection.Ki67 staining,transwell and ELISA were used to detect cell proliferation,migration and secretion of CCL2,CXCL8,IL-6 and VEGF-A.4.Cell transfection was used to down-regulate or up-regulate the expression of target circRNA in human fetal RPE cells.Ki-67 staining and transwell were used to detect cell proliferation and migration ability.Results:1.Microarray analysis revealed that 91 differentially expressed circRNAs between PVR-ERMs and secondary ERMs,including 55 up-regulated circRNAs and36 down-regulated circRNAs.qRT-PCRs verified 20 dysregulated circRNAs in the ERMs of PVR.Notably,circ₀043144 was the most up-regulated circRNA in PVR-ERMs.2.The expression of circ₀043144 in PVR-ERMs was significantly up-regulated as well as the PVR markers,such as PDGFA,PDGFC and KNG1.We also found that circ₀043144 was significantly up-regulated in serum and vitreous of PVR patients,and was positively correlated with the severity of PVR.3.Circ₀043144 silencing significantly decreased the proliferation and migration ability of ARPE-19,and decreased the secretion of cytokines and growth factors.And circ₀043144 overexpression significantly increased the proliferation and migration ability of ARPE-19,and decreased the secretion of cytokines and growth factors.4.Circ₀043144 silencing significantly decreased the proliferation and migration ability of human fetal RPE cells,and circ₀043144 overexpression significantly increased the proliferation and migration ability of human fetal RPE cells.Conclusion:This study indicates that circRNAs are potential regulators of the pathogenesis of PVR.Circ₀043144 is a promising prognostic and diagnostic indicator for PVR diseases.