Understanding Drivers of Ocular Fibrosis: Current and Future Therapeutic Perspectives

Mallone, Fabiana; Costi, Roberta; Marenco, Marco; Plateroti, Rocco; Minni, Antonio; Attanasio, Giuseppe; Artico, Marco; Lambìase, Alessandro

doi:10.3390/ijms222111748

Cited by 28 publications

(27 citation statements)

References 143 publications

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“…4A). Using wellcharacterized molecular markers [35][36][37][38] , we then grouped glia into resting, reactive, and fibrotic subgroups (Fig. 4B).…”

mentioning

confidence: 99%

Comparative analysis of single-cell and single-nucleus RNA-sequencing in a rabbit model of retinal detachment-related proliferative vitreoretinopathy

Santiago

Gimmen

et al. 2022

Preprint

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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 Muller glia were over-represented in snRNA-seq samples, while reactive Muller 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.

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“…4A). Using wellcharacterized molecular markers [35][36][37][38] , we then grouped glia into resting, reactive, and fibrotic subgroups (Fig. 4B).…”

mentioning

confidence: 99%

Comparative analysis of single-cell and single-nucleus RNA-sequencing in a rabbit model of retinal detachment-related proliferative vitreoretinopathy

Santiago

Gimmen

et al. 2022

Preprint

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“…Compared to other GSIs, DAPT and LY411575, two well studied GSIs in subretinal fibrosis and EMT, 45 RO displayed superiorities in its high efficiency and clinical translation. Among GSIs, DAPT, LY411575, and RO belong to the peptidomimetics-based GSIs, binding to an allosteric site in the γ-secretase complex and affecting its activity, which form the largest and most diverse set of GSIs.…”

Section: Discussionmentioning

confidence: 98%

RO4929097, a Selective γ-Secretase Inhibitor, Inhibits Subretinal Fibrosis Via Suppressing Notch and ERK1/2 Signaling in Laser-Induced Mouse Model

Zhang

Qin

Xie

et al. 2022

Invest. Ophthalmol. Vis. Sci.

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Purpose This study aimed to explore whether RO4929097 (RO), a specific γ-secretase inhibitor, could inhibit the subretinal fibrosis in laser-induced mouse model and the relevant molecular mechanisms. Methods Male C57BL/6J mice were used to produce choroidal neovascularization (CNV) and subretinal fibrosis by laser photocoagulation, and RO was administered intravitreally 1 day after laser induction. The sizes of CNV and subretinal fibrosis were measured and quantified in both 2D and 3D constructions. The ARPE-19 cell line and primary human RPE (phRPE) cells were treated with TGFβ1, in combination with or without RO, to examine Notch related molecules, epithelial mesenchymal transition (EMT), cell viability, migration, and contractile function, as well as the crosstalk between Notch and other EMT relevant signaling pathways. Results Intravitreal injection of RO reduced the sizes of both CNV and subretinal fibrosis in laser-induced young and old mice at day 7 and day 14 after laser induction. Moreover, EMT and Notch activation in RPE-choroid complexes from laser-induced mice were significantly attenuated by RO. In vitro, TGFβ1 activated Notch signaling and induced EMT in ARPE-19 cells, accompanied by enhanced EMT-related function, which were inhibited by RO. The inhibition of RO on EMT was further confirmed in TGFβ1-treated phRPE cells. Blockage of Notch signaling by RO could inhibit ERK1/2 signaling; whereas ERK1/2 inhibition had no effect on Notch. The action of RO was independent of Smad2/3 or p38, and co-inhibition of Notch and Smad2/3 showed synergistic effect on EMT inhibition. Conclusions RO exerts its antifibrotic effect by directly inhibiting Notch signaling and indirectly suppressing ERK1/2 signaling. Targeting Notch signaling might provide a therapeutic strategy in prevention and treatment of subretinal fibrosis in neovascular age-related macular degeneration (nAMD).

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“…Numerous factors are reported to be capable of driving EMT, such as hepatocyte growth factor (HGF), insulin-like growth factor-1 (IGF1), epidermal growth factor (EGF), platelet-derived growth factor (PDGF), and connective tissues growth factor (CTGF) [ 12 , 13 ]. The TGFB family is reported as a primary inducer [ 12 , 13 , 27 , 32 ]. It is found to cause increased myofibroblast-like epithelial cells in the lung, kidney, and breast, and it correlates with the above alterations in fibroproliferative diseases in the eye [ 4 , 27 , 32 , 33 ].…”

Section: Discussionmentioning

confidence: 99%

MiR-302d inhibits TGFB-induced EMT and promotes MET in primary human RPE cells

Binter

Hufendiek

et al. 2022

PLoS ONE

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Purpose Transforming growth factor-beta (TGFB)-mediated epithelial-mesenchymal transition (EMT) plays a crucial role in the pathogenesis of retinal fibrosis, which is one of the leading causes of impaired vision. Current approaches to treating retinal fibrosis focus, among other things, on inhibiting the TGFB signaling pathway. Transient expression of microRNAs (miRNAs) is one way to inhibit the TGFB pathway post-transcriptionally. Our previous study identified the miRNA miR-302d as a regulator of multiple TGFB-related genes in ARPE-19 cells. To further explore its effect on primary cells, the effect of miR-302d on TGFB-induced EMT in primary human retinal pigment epithelium (hRPE) was investigated in vitro. Methods hRPE cells were extracted from patients receiving enucleation. Transfection of hRPE cells with miR-302d was performed before or after TGFB1 stimulation. Live-cell imaging, immunocytochemistry staining, Western blot, and ELISA assays were utilized to identify the alterations of cellular morphology and EMT-related factors expressions in hRPE cells. Results hRPE cells underwent EMT by TGFB1 exposure. The transfection of miR-302d inhibited the transition with decreased production of mesenchymal markers and increased epithelial factors. Meanwhile, the phosphorylation of SMAD2 activated by TGFB1 was suppressed. Moreover, miR-302d expression promoted TGFB1-induced fibroblast-like hRPE cells to revert towards an epithelial stage. As confirmed by ELISA, miR-302d reduced TGFB receptor 2 (TGFBR2) and vascular endothelial growth factor A (VEGFA) levels 48 hours after transfection. Conclusions The protective effect of miR-302d might be a promising approach for ameliorating retinal fibrosis and neovascularization. MiR-302d suppresses TGFB-induced EMT in hRPE cells via downregulation of TGFBR2, even reversing the process. Furthermore, miR-302d reduces the constitutive secretion of VEGFA from hRPE cells.

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Understanding Drivers of Ocular Fibrosis: Current and Future Therapeutic Perspectives

Cited by 28 publications

References 143 publications

Comparative analysis of single-cell and single-nucleus RNA-sequencing in a rabbit model of retinal detachment-related proliferative vitreoretinopathy

Comparative analysis of single-cell and single-nucleus RNA-sequencing in a rabbit model of retinal detachment-related proliferative vitreoretinopathy

RO4929097, a Selective γ-Secretase Inhibitor, Inhibits Subretinal Fibrosis Via Suppressing Notch and ERK1/2 Signaling in Laser-Induced Mouse Model

MiR-302d inhibits TGFB-induced EMT and promotes MET in primary human RPE cells

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