There are many complex eye diseases which are the leading causes of blindness,however,the pathogenesis of the complex eye diseases is not fully understood,especially the underlying molecular mechanisms of N6-methylade...There are many complex eye diseases which are the leading causes of blindness,however,the pathogenesis of the complex eye diseases is not fully understood,especially the underlying molecular mechanisms of N6-methyladenosine(m6A)RNA methylation in the eye diseases have not been extensive clarified.Our review summarizes the latest advances in the studies of m6A modification in the pathogenesis of the complex eye diseases,including cornea disease,cataract,diabetic retinopathy,age-related macular degeneration,proliferative vitreoretinopathy,Graves’disease,uveal melanoma,retinoblastoma,and traumatic optic neuropathy.We further discuss the possibility of developing m6A modification signatures as biomarkers for the diagnosis of the eye diseases,as well as potential therapeutic approaches.展开更多
Purpose: To investigate the expression of thrombospondin 1 (TSP-1) in retinal pigment epithelium (RPE) and choroidal neovascular membranes (CNVMs) from patients with age-related macular degeneration (AMD). Methods: Ti...Purpose: To investigate the expression of thrombospondin 1 (TSP-1) in retinal pigment epithelium (RPE) and choroidal neovascular membranes (CNVMs) from patients with age-related macular degeneration (AMD). Methods: Tissue sections from normal human fetal and adult eyes and surgically removed CNVMs were immunostained for TSP-1 localization. Polymerase chain reaction and Western blotting were used to analyze TSP-1 mRNA and protein from human RPE cells, respectively. TSP-1 in the supernatant of cultured RPE cells and eye explants were measured using enzyme-linked immunosorbent assay. MTT assay was used to evaluate the RPE survival after TSP-1 treatment. Results: The strongest immunostaining for TSP-1 was observed in the RPE monolayer around drusen in early AMD. The intensity of TSP-1 staining in normal eye sections was much weaker than that of early AMD and CNVM. TSP-1 mRNA was positive in cultured fetal and adult RPE cells. There was increasing secretion of TSP-1 into the supernatant of cultured RPE and eye explants. The specific band of TSP-1 was identified by Western blot. No significant inhibition of RPE survival was found with the exposure to TSP-1. Conclusions: TSP-1 expression in drusen and CNVM was upregulated and associated with RPE monolayer. TSP-1 may be a natural negative regulator for choroidal neovascularization.展开更多
Purpose:To investigate the role of N6-methyladenosine(m^(6)A)RNA modification in the pathogenesis of Graves'ophthalmopathy(GO).Methods:Surgically excised extraocular muscles from 7 patients with GO and 5 subjects ...Purpose:To investigate the role of N6-methyladenosine(m^(6)A)RNA modification in the pathogenesis of Graves'ophthalmopathy(GO).Methods:Surgically excised extraocular muscles from 7 patients with GO and 5 subjects without GO were used.The global m^(6)A levels in the specimens were determined using an m^(6)A RNA methylation quantification kit.RNA sequencing(RNA-seq)was used to analyze the molecules involved in the regulation of m^(6)A RNA methylation and the differential expression of mRNAs between the two groups(4 eyes,respectively).The expression of m^(6)A RNA modification genes was evaluated by real-time PCR.The functional implications of the gene alterations between the GO and control specimens were determined by Gene Ontology analysis.Results:The m^(6)A level was significantly increased in the specimens of GO patients compared to the control specimens(P<0.05).The expression of m^(6)A methylation regulators,such as WT1 associated protein(WTAP),alkylation repair homolog protein 5(ALKBH5),E74 like ETS transcription factor 3(ELF3),YTH N6-methyladenosine RNA binding protein 2(YTHDF2),YTHDF3 and YTH domain containing 2(YTHDC2),was significantly upregulated(P<0.05).Gene Ontology enrichment analysis showed that the most highly upregulated genes and biological pathways were related to the immune response and inflammatory processes such as lymphocyte activation,leukocyte differentiation,cytokine production and cytokine-mediated signaling pathways.Conclusions:Our results suggest that m^(6)A methylation may play a critical role in the pathogenesis of GO and that targeting genes that regulate m^(6)A methylation may provide a new therapeutic approach for GO.展开更多
基金supported by the National Natural Science Foundation of China(No.81873681,2018,81770952,2017)the Basic Research Project of Henan Eye Hospital,China(No.20JCQN005,2020).
文摘There are many complex eye diseases which are the leading causes of blindness,however,the pathogenesis of the complex eye diseases is not fully understood,especially the underlying molecular mechanisms of N6-methyladenosine(m6A)RNA methylation in the eye diseases have not been extensive clarified.Our review summarizes the latest advances in the studies of m6A modification in the pathogenesis of the complex eye diseases,including cornea disease,cataract,diabetic retinopathy,age-related macular degeneration,proliferative vitreoretinopathy,Graves’disease,uveal melanoma,retinoblastoma,and traumatic optic neuropathy.We further discuss the possibility of developing m6A modification signatures as biomarkers for the diagnosis of the eye diseases,as well as potential therapeutic approaches.
文摘Purpose: To investigate the expression of thrombospondin 1 (TSP-1) in retinal pigment epithelium (RPE) and choroidal neovascular membranes (CNVMs) from patients with age-related macular degeneration (AMD). Methods: Tissue sections from normal human fetal and adult eyes and surgically removed CNVMs were immunostained for TSP-1 localization. Polymerase chain reaction and Western blotting were used to analyze TSP-1 mRNA and protein from human RPE cells, respectively. TSP-1 in the supernatant of cultured RPE cells and eye explants were measured using enzyme-linked immunosorbent assay. MTT assay was used to evaluate the RPE survival after TSP-1 treatment. Results: The strongest immunostaining for TSP-1 was observed in the RPE monolayer around drusen in early AMD. The intensity of TSP-1 staining in normal eye sections was much weaker than that of early AMD and CNVM. TSP-1 mRNA was positive in cultured fetal and adult RPE cells. There was increasing secretion of TSP-1 into the supernatant of cultured RPE and eye explants. The specific band of TSP-1 was identified by Western blot. No significant inhibition of RPE survival was found with the exposure to TSP-1. Conclusions: TSP-1 expression in drusen and CNVM was upregulated and associated with RPE monolayer. TSP-1 may be a natural negative regulator for choroidal neovascularization.
基金This work was supported by grants from the National Natural Science Foundation of China(grant nos.81770943 and 81873681)the Beijing Municipal Science and Technology Commission(Capital Char-acteristic Clinic Applied Research Project,Z161100000516037).
文摘Purpose:To investigate the role of N6-methyladenosine(m^(6)A)RNA modification in the pathogenesis of Graves'ophthalmopathy(GO).Methods:Surgically excised extraocular muscles from 7 patients with GO and 5 subjects without GO were used.The global m^(6)A levels in the specimens were determined using an m^(6)A RNA methylation quantification kit.RNA sequencing(RNA-seq)was used to analyze the molecules involved in the regulation of m^(6)A RNA methylation and the differential expression of mRNAs between the two groups(4 eyes,respectively).The expression of m^(6)A RNA modification genes was evaluated by real-time PCR.The functional implications of the gene alterations between the GO and control specimens were determined by Gene Ontology analysis.Results:The m^(6)A level was significantly increased in the specimens of GO patients compared to the control specimens(P<0.05).The expression of m^(6)A methylation regulators,such as WT1 associated protein(WTAP),alkylation repair homolog protein 5(ALKBH5),E74 like ETS transcription factor 3(ELF3),YTH N6-methyladenosine RNA binding protein 2(YTHDF2),YTHDF3 and YTH domain containing 2(YTHDC2),was significantly upregulated(P<0.05).Gene Ontology enrichment analysis showed that the most highly upregulated genes and biological pathways were related to the immune response and inflammatory processes such as lymphocyte activation,leukocyte differentiation,cytokine production and cytokine-mediated signaling pathways.Conclusions:Our results suggest that m^(6)A methylation may play a critical role in the pathogenesis of GO and that targeting genes that regulate m^(6)A methylation may provide a new therapeutic approach for GO.
