Comparative analysis of single-cell and single-nucleus RNA-sequencing in a rabbit model of retinal detachment-related proliferative vitreoretinopathy
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PROVIDER: PRJNA898403 | ENA |
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Project description:Proliferative vitreoretinopathy (PVR) is the most common cause of failure of retinal reattachment surgery and the molecular changes leading to this aberrant wound healing process is currently unknown. We aimed to study PVR pathogenesis using single-cell transcriptomics to dissect cellular heterogeneity in a rabbit PVR model. PVR was induced unilaterally in Dutch Belted rabbits. At different timepoints following PVR induction, retinas were dissociated into either cells or nuclei suspension and processed for single-cell or single-nucleus RNA sequencing (scRNA-seq or snRNA-seq). ScRNA-Seq and snRNA-Seq were conducted on retinas at 4 hours and 14 days after disease induction. While the capture rate of unique molecular identifiers (UMI) and genes were greater in scRNA-seq samples, overall gene expression profiles of individual cell types were highly correlated between scRNA-seq and snRNA-seq. A major disparity between the two sequencing modalities is the cell type capture rate, however, with glial cell types over-represented in scRNA-seq, and inner retinal neurons were enriched by snRNA-seq. Furthermore, fibrotic Müller glia were over-represented in snRNA-seq samples, while reactive Müller glia were in scRNA-seq samples. Trajectory analyses were similar between the two methods, allowing for the combined analysis of the scRNA-seq and snRNA-seq datasets.These findings highlight limitations of both scRNA-seq and snRNA-seq analysis and imply that use of both techniques can more accurately identify transcriptional networks critical for aberrant fibrogenesis in PVR.
2022-11-08 | GSE217333 | GEO
Project description:Proliferative vitreoretinopathy (PVR) is a severe vision-threatening disorder characterized by the formation of cicatricial fibrous membranes leading to traction retinal detachment. As the pathophysiology remains unclear, there has been no effective therapeutic approach to date other than vitreoretinal surgery.
2012-09-21 | GSE41019 | GEO
Project description:scRNAseq of mouse proliferative vitreoretinopathy
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Project description:Transcriptional Characterisation of Proliferative Vitreoretinopathy
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Project description:We performed single cell transcriptional profiling of mouse proliferative vitreoretinopathy (PVR), to decipher the molecular mechanisms underlying mouse PVR fibrogenesis. In this study, through combination of the single-cell transcriptional profiling and genetic tracing, we provided comprehensive discrimination for the sources of PVR-associated fibroblasts during ocular wound healing.
2025-09-05 | GSE274480 | GEO
Project description:Fibrous membrane (FM), the hallmark for proliferative diabetic retinopathy (PDR) and proliferative vitreoretinopathy (PVR), can cause hemorrhages and retinal detachment, which may lead to blindness if not properly treated. However, little is known about the pathophysiology of FM. In this study, we successfully employed single-cell RNA sequencing on the small-sized vitreous FMs, and generated a comprehensive cell atlas of FMs derived from PVR and PDR. Distinct cell compositions were identified in the FMs, with microglia as the major cell population. Additionally, our analysis revealed a spectrum of microglia activation states with distinct molecular signatures and intercellular interactions in disease-specific microenvironment. This description of microglia phenotypes in the FM of PVR and PDR may offer insight into the activation of microglia in the FM pathogenesis, as well as potential signaling pathways amenable to disease-specific intervention.
2022-01-27 | GSE165784 | GEO
Project description:Proliferative vitreoretinopathy (PVR) is a severe vision-threatening disorder characterized by the formation of cicatricial fibrous membranes leading to traction retinal detachment. As the pathophysiology remains unclear, there has been no effective therapeutic approach to date other than vitreoretinal surgery. In order to identify genes associated with the pathogenesis of PVR, we performed gene expression analyses in fibrous membrane in patients with PVR using DNA microarray technology. This study was approved by the Ethics Committee of the Kyushu University Hospital and Fukuoka University Chikushi Hospital, and the surgical specimens were handled in accordance with the ethical standards of the 1989 Declaration of Helsinki. All patients gave informed consent before inclusion in the study. Fibrousl membranes were surgically dissected from the retinal surface with horizontal scissors of patients with PVR undergoing pars plana vitrectomy. Total RNA were extracted from the fibrous membranes with three different PVR patients. RNA from human retina was obtained from Clontech (Palo Alto, CA).
2012-09-21 | E-GEOD-41019 | biostudies-arrayexpress
Project description:Purpose: The development of vitreoretinal scars remains an unsolved challenge in clinical practice and often leads to repeated revision surgery and blindness due to lack of efficient therapy. The aim of this study was to characterize the cellular and molecular environment in vitreoretinal scar tissue from patients with proliferative vitreoretinopathy (PVR) in comparison to membranes of the vitreoretinal junction, in order to subsequently identify potential drug treatment options using bioinformatics techniques. Methods: A total of 23 patients undergoing vitrectomy for retinal detachment due to proliferative vitreoretinopathy (PVR, n = 15), idiopathic macular hole (MH, n = 10) or idiopathic macular pucker (MP, n = 8), were included in this study. Vitreoretinal samples were analysed by RNA sequencing, MS-CyTOF proteomic and by conventional immunohistochemistry. The cellular microenvironment was assessed by bioinformatics cell type enrichment analysis. Drug target screening was performed by using a transcriptome-based drug-repurposing approach using drug-exposure transcriptome data from public available databases. Results: RNA sequencing of vitreoretinal tissue samples showed distinct transcriptional differences of PVR, ERM and ILM samples allowing an accurate clustering according to the tissue types. The cellular microenvironment of PVR membranes was characterized by the enrichment of melanocytes, astrocytes and various immune cell types, such as M2 macrophages. Differential gene expression (DEG) analysis revealed XX up- and XX downregulated genes in PVR compared to ILM. Among them, FN1 (fibronectin1), SPARC (osteonectin) and various collagens were among the most upregulated factors in PVR, contributing to biological processes such as the organization of extracellular structures, the regulation of cell adhesion and the morphogenesis of blood vessels. Finally, we identified 13 drugs that induce a gene expression profile complementary to the PVR signature and thus represent potential therapeutic options for PVR, including aminocaproic acid and various topoisomerase 2A inhibitors such as doxorubicin, etoposide and mitoxantrone. Conclusion: The present study reveals significant differences in the transcriptional signature of vitreoretinal scar tissue including PVR, MP and ILM tissue. Among a plethora of differentially expressed genes, FN1 and SPARC appeared to be central mediators underlying PVR development. Based on the transcriptome analyses, aminocaproic acid and topoisomerase-2 inhibitors, such as doxorubicin, etoposide and mitoxantrone, were identified as compatible drugs for the treatment of PVR.
2022-05-25 | GSE179603 | GEO
Project description:To characterize vitreous humor (VH) exosomes and to explore their role in the development of proliferative vitreoretinopathy (PVR) using mass-spectrometry based proteome profiling.
2022-06-09 | PXD028539 | Pride
Project description:PurposeProliferative vitreoretinopathy (PVR) is the most common cause of failure of retinal reattachment surgery, and the molecular changes leading to this aberrant wound healing process are currently unknown. Our ultimate goal is to study PVR pathogenesis by employing single-cell transcriptomics to dissect cellular heterogeneity.DesignHere we aimed to compare single-cell RNA sequencing (scRNA-seq) and single-nucleus RNA-sequencing (snRNA-seq) of retinal PVR samples in the rabbit model.ParticipantsUnilateral induction of PVR lesions in rabbit eyes with contralateral eyes serving as controls.MethodsProliferative vitreoretinopathy was induced unilaterally in Dutch Belted rabbits. At different timepoints after PVR induction, retinas were dissociated into either cells or nuclei suspension and processed for scRNA-seq or snRNA-seq.Main outcome measuresSingle cell and nuclei transcriptomic profiles of retinas after PVR induction.ResultsSingle-cell RNA sequencing and snRNA-seq were conducted on retinas at 4 hours and 14 days after disease induction. Although the capture rate of unique molecular identifiers and genes were greater in scRNA-seq samples, overall gene expression profiles of individual cell types were highly correlated between scRNA-seq and snRNA-seq. A major disparity between the 2 sequencing modalities was the cell type capture rate, however, with glial cell types overrepresented in scRNA-seq, and inner retinal neurons were enriched by snRNA-seq. Furthermore, fibrotic Müller glia were overrepresented in snRNA-seq samples, whereas reactive Müller glia were overrepresented in scRNA-seq samples. Trajectory analyses were similar between the 2 methods, allowing for the combined analysis of the scRNA-seq and snRNA-seq data sets.ConclusionsThese findings highlight limitations of both scRNA-seq and snRNA-seq analysis and imply that use of both techniques together can more accurately identify transcriptional networks critical for aberrant fibrogenesis in PVR than using either in isolation.Financial disclosuresProprietary or commercial disclosure may be found after the references.
| S-EPMC10365955 | biostudies-literature