Project description:As the most prevalent, abundant and conserved internal cotranscriptional modification in eukaryotic RNAs, N6-methyladenosine (m6A) plays vital role in the progression of different types of human diseases, including cancers. m6A RNA methylation have been reported to affects the physiological and pathological progressions of hematopoietic, central nervous and reproductive systems. As diabetic retinopathy is one of the most importent microvascular complication of diabetes, in this work, MeRIP-seq was employed to investigate the RNA m6A levels in the vitreous body samples of diabetic retinopathy and control groups. A group of mRNAs were suggested with significantly fluctuated transcription and m6A levels, and they would be inverstigated in the further work to understand the mechanisam of the progression of diabetic retinopathy.
Project description:We identified lncRNA expression profiles in vitreous samples between proliferative diabetic retinopathy (PDR) patients and idiopathic macular hole (IMH) patients, and between PDR patients who had received preoperative anti-vascular endothelial growth factor (anti-VEGF) therapy and PDR patients who had received surgery alone. There had been the systemic expression differences in vitreous at the microarray level from PDR patients and IMH patients, and from PDR patients after anti-VEGF treatment and untreated PDR patients.
Project description:To reveal the expression profiles of transfer RNA-derived small RNA (tsRNA)s and microRNA (miRNA)s in the vitreous humour of proliferative diabetic retinopathy (PDR).
Project description:Despite great advancements in proliferative diabetic retinopathy (PDR) therapy over the last decades, one third of treated patients continue to lose vision. Vitreous macrophages called hyalocytes have been implied in the pathogenesis of vitreoretinal proliferative disease previously but little is known about their role in PDR. In this study, we address molecular and cellular alterations of the PDR vitreous, in order to assess the potential contribution of hyalocytes to PDR development. A total of 55 patients were included in this study encompassing RNA-Sequencing analysis of vitreous hyalocytes from PDR and control patients, multiplex immunoassay and ELISA analysis of undiluted vitreous samples and isolation and immunohistochemical staining of cultured porcine hyalocytes. Our transcriptional analysis revealed an enhanced inflammatory signature of hyalocytes contributing to the cytokine pool within the PDR vitreous by expressing, among others, interleukin-6. The data further indicate an expression shift, involving angiopoietin-2, in PDR hyalocytes converting them to a proangiogenic cellular component, which may mediate formation of retinal neovascularization, hallmark of PDR. Finally, our data suggest a role of vitreous macrophages in erythrophagocytosis and, thereby, removal of vitreous hemorrhage, a severe complication of PDR. Immunomodulation of hyalocytes may thus prove an essential novel therapeutical approach in diabetic vitreoretinal disease.
Project description:To investigate the key regulators of the disease by comparing the abundance of vitreous proteins between the patients with proliferative diabetic retinopathy (PDR) and the controls with idiopathic epiretinal membrane (iERM).
Project description:The microRNA profiles in the vitreous of proliferative vitreoretinal disease (PVD) such as proliferative diabetic retinopathy with fibrovascular membrane and macular hole (MH) patients were studied by RT-PCR. From each individual in the two cohorts: the PVD (n=3) and MH patients (n=3), vitreous specimens were collected and microRNAs were extracted for miRNA profiles analysis.
Project description:The microRNA profiles in the vitreous of proliferative vitreoretinal disease (PVD) such as proliferative diabetic retinopathy with fibrovascular membrane and macular hole (MH) patients were studied by RT-PCR.
Project description:Diabetic retinopathy (DR) is a major complication of diabetes mellitus causing significant vision loss. Despite the success of anti-VEGF therapy as an effective therapy, many DR patients do not respond well to the treatment, emphasizing the involvement of other molecular players. To unveil these, we performed deep vitreous proteome profiling of patients with proliferative DR and extracted aberrated protein networks. This revealed impairment in ectodomain shedding of several transmembrane proteins playing critical roles in neurodegeneration and angiogenesis, that pointed to defects in their regulating sheddases, particularly ADAM10, which emerged as the predominant sheddase. We confirmed that ADAM10 protease activity was reduced in ocular disease models and established that activation of ADAM10 can suppress endothelial cell activation and angiogenesis. Furthermore, we identified impaired ADAM10-AXL axis as a retinal angiogenic driver. Taken together, we demonstrate restoration of aberrant ectodomain shedding as an effective strategy for treating DR and propose ADAM10 as an attractive novel target.
Project description:PURPOSE: To investigate the circulatory microRNA (miRNA) profiles of aqueous, vitreous, and plasma in order to identify biomarkers in aqueous humor or plasma that are reflecting changes in vitreous of patients with diabetes. METHODS: Aqueous, vitreous and plasma samples were collected from a total of 27 patients - 11 controls (macular pucker or macular hole patients) and 16 patients with diabetes mellitus (DM) undergoing vitreoretinal surgery: DM-Type I with proliferative diabetic retinopathy (PDR) (DMI-PDR), DM Type II with PDR (DMII-PDR) and DM Type II with nonproliferative DR (DMII-NPDR). MiRNAs were isolated using Qiagen microRNeasy kit, quantified on BioAnalyzer, labeled with FlashTag kit, and profiled on Affymetrix GeneChip miRNA 3.0 microarrays. Data analysis was done using Expression Console (EC), Transcriptome Analysis Console (TAC), and Ingenuity Pathway Analysis (IPA) software. RESULTS: Our comparison of circulatory miRNA population of aqueous and vitreous humor and plasma showed that out of total of 847 human miRNA probes on the Affymetrix GeneChip miRNA 3.0 we found common miRNAs for both aqueous and vitreous samples, as well as larger number of unique miRNA, dependent on the DM type and presence of retinopathy. Most of the dysregulated miRNAs in aqueous and vitreous of DM patients were upregulated, while in plasma, most of the DM-specific miRNAs were downregulated. Dysregulation of miRNAs in aqueous generally do not appear to be a good representative of the miRNA abundance in vitreous, or plasma, although we did identify a few candidates for common biomarkers: let-7b, miR-320b, miR-762 and miR-4488. Additionally, each of the DR subtypes showed a set of miRNA that is uniquely dysregulated in each fluid, for example in aqueous samples for DMII-NPDR it was miR-455-3p, for DMII-PDR was miR-296, and for DMI-PDR it was miR-3202. Pathway analysis identified TGF-beta and VEGF pathways as the common targets for miRNAs dysregulated in DR aqueous and vitreous. CONCLUSIONS: The comparative profiling of circulatory miRNAs in aqueous, vitreous, and plasma showed that a small number of circulatory miRNAs displayed differential presence in controls vs. diabetic retinopathy. A pattern is emerging of sets of miRNA that are common or uniquely dysregulated in the blood plasma or ocular fluids of DR subtypes, offering promise for the use of ocular fluids and plasma for identifying diagnostic and therapeutic targets.