Chen J.-F.,Wenzhou University |
Chen J.-F.,State Key Laboratory Cultivation Base and Key Laboratory of Vision Science |
Zhang S.,Wenzhou University |
Zhang S.,State Key Laboratory Cultivation Base and Key Laboratory of Vision Science |
And 7 more authors.
Molecular Aspects of Medicine | Year: 2017
Retinopathy of prematurity (ROP) is a major cause of childhood blindness in the world and is caused by oxygen-induced damage to the developing retinal vasculature, resulting in hyperoxia-induced vaso-obliteration and subsequent delayed retinal vascularization and hypoxia-induced pathological neovascularization driven by vascular endothelial growth factor (VEGF) signaling pathway in retina. Current anti-VEGF therapy has shown some effective in a clinical trial, but is associated with the unintended effects on delayed eye growth and retinal vasculature development of preterm infants. Notably, cellular responses to hypoxia are characterized by robust increases in extracellular adenosine production and the markedly induced adenosine receptors, which provide a novel target for preferential control of pathological angiogenesis without affecting normal vascular development. Here, we review the experimental evidence in support of adenosine receptor-based therapeutic strategy for ROP, including the aberrant adenosine signaling in oxygen-induced retinopathy and the role of three adenosine receptor subtypes (A1R, A2AR, A2BR) in development and treatment of ROP using oxygen-induced retinopathy models. The clinical and initial animal evidence that implicate the therapeutic effect of caffeine (a non-selective adenosine receptor antagonist) in treatment of ROP are highlighted. Lastly, we discussed the translational potential as well therapeutic advantage of adenosine receptor- and caffeine-based therapy for ROR and possibly other proliferative retinopathy. © 2017 Elsevier Ltd.
PubMed | State Key Laboratory Cultivation Base and Key Laboratory of Vision Science, Boston University, Wenzhou University and Georgia Regents University
Type: Journal Article | Journal: Investigative ophthalmology & visual science | Year: 2016
We critically evaluated the role of the adenosine A1 receptor (A1R) in normal development of retinal vasculature and pathogenesis of retinopathy of prematurity (ROP) by using the A1R knockout (KO) mice and oxygen-induced retinopathy (OIR) model.Mice deficient in A1Rs and their wild-type (WT) littermates were examined during normal postnatal development or after being subjected to 75% oxygen from postnatal day (P) 7 to P12 and to room air from P12 to P17 (OIR model of ROP). Retinal vascularization was examined by whole-mount fluorescence and cross-sectional hematoxylin-eosin staining. Cellular proliferation, astrocyte and microglial activation, and tip cell function were determined by isolectin staining and immunohistochemistry. Apoptosis was determined by TUNEL assay.Genetic deletion of the A1R did not affect normal retinal vascularization during postnatal development with indistinguishable three-layer vascularization patterns in retina between WT and A1R KO mice. In the OIR model, genetic deletion of the A1R resulted in stage-specific effects: reduced hyperoxia-induced retinal vaso-obliteration at P12, but reduced avascular area and attenuated hypoxia-induced intraretinal revascularization without affecting intravitreal neovascularization at P17 and reduced avascular areas in retina at P21. These distinct effects of A1Rs on OIR were associated with A1R control of apoptosis mainly in inner and outer nuclear layers at the vaso-obliterative phase (P12) and the growth of endothelium tip cells at the vasoproliferative phase (P17), without modification of cellular proliferation, astrocytic activation, and tissue inflammation.Adenosine A1 receptor activity is not required for normal postnatal development of retinal vasculature but selectively controls hyperoxia-induced vaso-obliteration and hypoxia-driven revascularization by distinct cellular mechanisms.
Chen X.,Wenzhou Medical College |
Chen X.,State Key Laboratory Cultivation Base and Key Laboratory of Vision Science |
He D.,Wenzhou Medical College |
He D.,State Key Laboratory Cultivation Base and Key Laboratory of Vision Science |
And 14 more authors.
Investigative Ophthalmology and Visual Science | Year: 2013
MicroRNA-124a (miR-124a), an abundant microRNA in the central neuron system, has been linked to tumor progression. Here, we investigated the role of miR-124a in uveal melanoma development. METHODS. Expression of miR-124a in uveal melanoma cells was examined using real time RT-PCR. The effect of miR-124a on cell proliferation, migration, and invasion was analyzed using MTS assay, flow cytometry, and transwell experiments. The ability of miR-124a to repress tumor growth was tested in vivo. Target genes of miR-124a were first predicted by bioinformatics, confirmed using a luciferase assay, and their expression determined by Western blotting. DNA methylation and histone modification of miR-124a was analyzed by methylation-specific PCR and ChIP assay. Finally, epigenetic drugs were used to alter the expression of miR-124a. RESULTS. miR-124a expression was downregulated in both uveal melanoma cells and clinical specimens. Transient transfection of miR-124a into uveal melanoma cells inhibited cell growth, migration, and invasion. Moreover, introduction of miR-124a suppressed in vivo growth of tumor. Potential targets of miR- 124a were found to include CDK4, CDK6, cyclin D2, and EZH2. Knockdown of EZH2 by siRNA resulted in inhibition of uveal melanoma cell migration and invasion. In addition, miR- 124a expression was found to be regulated via epigenetic mechanisms, with its expression restored when cells were treated with a DNA hypomethylating agent, 5-aza-20-deoxycytidine, and a histone deacetylase inhibitor, trichostatin A. CONCLUSIONS. Our results demonstrated that miR-124a could function as a potent tumor suppressor by regulation of multiple targets, and was epigenetically silenced in the development of uveal melanoma. © 2013 The Association for Research in Vision and Ophthalmology, Inc.
Yan D.,Wenzhou Medical College |
Yan D.,State Key Laboratory Cultivation Base and Key Laboratory of Vision Science |
da Dong X.,Columbia University |
Chen X.,Wenzhou Medical College |
And 15 more authors.
PLoS ONE | Year: 2012
MicroRNAs (miRNAs) are endogenous small non-coding RNAs that play central roles in diverse pathological processes. In this study, we investigated the effect of microRNA-182 (miR-182) on the development of posterior uveal melanomas. Initially, we demonstrated that miR-182 expression was dependent on p53 induction in uveal melanoma cells. Interestingly, transient transfection of miR-182 into cultured uveal melanoma cells led to a significant decrease in cell growth, migration, and invasiveness. Cells transfected with miR-182 demonstrated cell cycle G1 arrest and increased apoptotic activity. Using bioinformatics, we identified three potential targets of miR-182, namely MITF, BCL2 and cyclin D2. miR-182 was shown to have activity on mRNA expression by targeting the 3′ untranslated region of MITF, BCL2 and cyclin D2. Subsequent Western blot analysis confirmed the downregulation of MITF, BCL2 and cyclin D2 protein expression. The expression of oncogene c-Met and its downstream Akt and ERK1/2 pathways was also downregulated by miR-182. Concordant with the findings that miR-182 was decreased in uveal melanoma tissue samples, overexpression of miR-182 also suppressed the in vivo growth of uveal melanoma cells. Our results demonstrated that miR-182, a p53 dependent miRNA, suppressed the expression of MITF, BCL2, cyclin D2 and functioned as a potent tumor suppressor in uveal melanoma cells. © 2012 Yan et al.
An J.,Wenzhou University |
An J.,State Key Laboratory Cultivation Base and Key Laboratory of Vision Science |
Chen X.,Wenzhou University |
Chen X.,State Key Laboratory Cultivation Base and Key Laboratory of Vision Science |
And 10 more authors.
Investigative Ophthalmology and Visual Science | Year: 2015
Purpose. MicroRNAs (miRNAs) are endogenous short chain (∼22-nucleotide) noncoding RNAs that inhibit protein translation through binding to target mRNAs. Recent studies have implied that miRNAs play a regulatory role in corneal development. Here we profile their involvement in corneal epithelial renewal, develop an miRNA-target network that affects wound healing outcome, and investigate the function of miR-204 in this response. Methods. NanoString nCounter technology and bioinformatics analyzed miRNA expression levels and their targets during mouse corneal epithelial wound healing. Real-time RT-PCR was performed to detect miR-204 expression in mouse corneal epithelium. Human corneal epithelial cells (HCECs) were transfected with miR-204 using transfection reagent. MTS (3-[4,5-dimethylthiazol-2-yl]-5-[3-carboxymethoxyphenyl]-2-[4-sulfophenyl]-2H-tetrazolium, inner salt) and a scratch wound-healing assay evaluated the effects of miR-204 expression on HCEC proliferation and migration, respectively. Cell cycle analysis was performed by flow cytometry. Expression of sirtuin 1 (SIRT1) was determined by Western blot analysis. Results. Fifteen miRNAs were dramatically downregulated, whereas 14 other miRNAs were markedly upregulated during corneal wound healing. Expression of miR-204 fell the most during this process. Transfection of miR-204 into HCECs led to a significant decline in cell proliferation and induced cell cycle G1-arrest. Furthermore, in these cells, miR-204 also inhibited migration. Sirtuin 1 was confirmed as a target of miR-204. Conclusions. During mouse corneal epithelial wound healing, a complex miRNA-gene network was resolved that is modulated by changes in miR-204 expression. Downregulation of this miRNA appears to be an essential response to injury since its decline promotes human corneal epithelial cell proliferation and migration. Therefore, miR-204 could be a biomarker of this process. © 2015 The Association for Research in Vision and Ophthalmology, Inc.
Huang X.-F.,Wenzhou University |
Huang X.-F.,State Key Laboratory Cultivation Base and Key Laboratory of Vision Science |
Wu J.,Wenzhou University |
Wu J.,State Key Laboratory Cultivation Base and Key Laboratory of Vision Science |
And 6 more authors.
Genetics in Medicine | Year: 2015
Purpose:Retinitis pigmentosa (RP) is a major cause of heritable human blindness with extreme genetic heterogeneity. A large number of causative genes have been defined by next-generation sequencing (NGS). However, due to technical limitations, determining the existence of uncovered or low-depth regions is a fundamental challenge in analyzing NGS data. Therefore, undetected mutations may exist in genomic regions less effectively covered by NGS.Methods:To address this problem, we tested a complementary approach for identifying previously undetected mutations in NGS data sets. The strategy consisted of coverage-based analysis and additional target screening of low-depth regions. Fifty RP patients were analyzed, and none of the mutations found had previously been identified by NGS.Results:Coverage-based analysis indicated that, because of a highly repetitive sequence, the RPGR open reading frame (ORF)15 was located in an uncovered or low-depth region. Through additional screening of ORF15, we identified pathogenic mutations in 14% (7/50) of patients, including four novel mutations first described herein.Conclusion:In brief, we support the need for a complementary approach to identify mutations undetected by NGS, underscoring the power and significance of combining coverage-based analysis with additional target screening of low-depth regions in improving diagnosis of genetic diseases. In addition to its usefulness in RP, this approach is likely applicable to other Mendelian diseases.Genet Med 17 4, 307-311.
Hou Q.,Wenzhou University |
Hou Q.,State Key Laboratory Cultivation Base and Key Laboratory of Vision Science |
Tang J.,Wenzhou University |
Tang J.,State Key Laboratory Cultivation Base and Key Laboratory of Vision Science |
And 11 more authors.
Investigative Ophthalmology and Visual Science | Year: 2013
Purpose. Retinal pigment epithelial (RPE) cells play important roles in ophthalmologic diseases such as proliferative vitreoretinopathy, AMD, and diabetic retinopathy. MicroRNA-34a (miR-34a) has been reported to be important in the regulation of cell proliferation, migration, differentiation, and apoptosis. In this study, we explored the effects of miR-34a on RPE cells. Methods. The expression level of miR-34a in subconfluent and postconfluent ARPE-19 cells was investigated with quantitative real-time PCR. MicroRNA mimic and small interfering RNA (siRNA) were transiently transfected into RPE cells. Transfected RPE cells were analyzed with WST-1 proliferation assay, and their migration was analyzed with transwell assay and in vitro scratch study. The expression or activation of target proteins was detected by Western blotting. Results. MicroRNA-34a was significantly downregulated in subconfluent ARPE-19 cells compared with postconfluent cells. Introduction of miR-34a inhibited the proliferation and migratory ability of RPE cells without obvious cell apoptosis. In miR-34a transfected cells, many important proliferation and/or migration related molecules such as c-Met, CDK2, CDK4, CDK6, E2F1, and phosphorylated-Cdc2 (p-Cdc2) were downregulated. Small interfering RNA designed to target c-Met also inhibited the proliferation and migration of RPE cells and downregulated CDK2, CDK6, E2F1, and p-Cdc2. Conclusions. MicroRNA-34a is downregulated in subconfluent RPE cells. MicroRNA-34a can inhibit the proliferation and migration of RPE cells through downregulation of its targets c-Met and other cell cycle-related molecules. Our results indicated that miR-34a is involved in the regulation of RPE cells. © 2013 The Association for Research in Vision and Ophthalmology, Inc.
PubMed | Chinese Academy of Sciences, State Key Laboratory Cultivation Base and Key Laboratory of Vision Science, Wenzhou University, Changzhi Medical College and U.S. National Institutes of Health
Type: Case Reports | Journal: Genetics in medicine : official journal of the American College of Medical Genetics | Year: 2015
Genetic etiology of congenital/infantile nystagmus remains largely unknown. This study aimed to identify genomic mutations in patients with infantile nystagmus and an associated disease network.Patients with inherited and sporadic infantile nystagmus were recruited for whole-exome and Sanger sequencing. -Mannosidase activities were measured. Gene expression, protein-protein interaction, and nystagmus-associated lysosomal storage disease (LSD) genes were analyzed.A novel heterozygous mutation (c.2013G>A; p.R638H) of MANBA, which encodes lysosomal -mannosidase, was identified in patients with autosomal-dominant nystagmus. An additional mutation (c.2346T>A; p.L749H) in MANBA was found by screening patients with sporadic nystagmus. MANBA was expressed in the pretectal nucleus of the developing midbrain, known to be involved in oculomotor and optokinetic nystagmus. Functional validation of these mutations demonstrated a significant decrease of -mannosidase activities in the patients as well as in mutant-transfected HEK293T cells. Further analysis revealed that nystagmus is present in at least 24 different LSDs involving the brain.This is the first identification of MANBA mutations in patients with autosomal-dominant nystagmus, suggesting a new clinical entity. Because -mannosidase activities are required for development of the oculomotor nervous system, our findings shed new light on the role of LSD-associated genes in the pathogenesis of infantile nystagmus.
PubMed | State Key Laboratory Cultivation Base and Key Laboratory of Vision Science, Wenzhou University and Zhejiang University
Type: Journal Article | Journal: PloS one | Year: 2016
As increases in hepatocyte growth factor/scatter factor (HGF/SF) induce retinal pigment epithelial (RPE) migration and proliferation into the vitreous cavity and contribute to proliferative vitreoretinopathy (PVR) development, we determined if changes in miR-182 expression affect such behavioral changes. We found that miR-182 expression was less in PVR clinical samples than in primary RPE cells whereas c-Met was upregulated. Ectopic miR-182 inhibited RPE cell proliferation, cell cycle, and migration. Bioinformatic analysis identified c-Met as a miR-182 target, which was confirmed with the luciferase reporter assay. Transfection of miR-182 into RPE cells induced c-Met downregulation, which led to reduced cell proliferation and migration through declines in p-Akt formation. MiR-182 downregulation along with c-Met upregulation in PVR tissues suggest that these two opposing effects play important roles in PVR development. As ectopic miR-182 expression suppressed RPE cell proliferation and migration, strategies to selectively upregulate miR-182 expression in a clinical setting may provide a novel option to treat this disease.
Chen W.,Wenzhou Medical College |
Chen W.,State Key Laboratory Cultivation Base and Key Laboratory of Vision Science |
Chen J.,Wenzhou Medical College |
Chen J.,State Key Laboratory Cultivation Base and Key Laboratory of Vision Science |
And 4 more authors.
American Journal of Ophthalmology | Year: 2013
Purpose: To assess correlations between associated factors and treatment outcome of isoametropic amblyopia attributable to high hyperopia in children and to evaluate changes in retinal thickness during amblyopic treatment. Design: Retrospective (visual outcome) and prospective (retinal thickness) study. Methods: Isoametropic (spherical equivalent ≥5.00 diopters in both eyes) amblyopic subjects (n = 217) with corrected visual acuity (VA) ≤0.5 (20/40) for children <5 years old and VA ≤0.6 (20/32) for children 6-10 years old were included. Sixty-nine of these children had refractive accommodative esotropia. All subjects were treated by full-time wearing of optical correction. The magnitude of spherical equivalent hyperopia, age at first treatment, VA, treatment duration, and binocular alignment were analyzed. Thicknesses of the retinal macula and nerve fiber layer were measured using optical coherence tomography in selected amblyopic children (n = 27) during treatment and in control subjects (n = 31). Results: The mean follow-up was 28.8 months. The age at first optical correction ranged from 3 to 10 years. The mean VA improved from 0.38 to 0.82, with 74.7% having acuity ≥0.8 and 28.6% having acuity ≥1.0. VA improvement was positively correlated with duration of treatment and negatively correlated with age at first correction. Foveal thickness slightly decreased after treatment; however, it was not correlated with visual improvement. Conclusion: Visual acuity of isoametropic amblyopia improved satisfactorily with spectacle correction and vision therapy. Treatment duration had the greatest impact on VA improvement. Age at first correction also influenced VA improvement but was not a good clinical predictor. Foveal thinning occurring with treatment was not correlated with visual improvement. © 2013 by Elsevier Inc. All rights reserved.