Experimental Models to Study Epithelial-Mesenchymal Transition in Proliferative Vitreoretinopathy

Datlibagi, Azine; Zein-El-Din, Anna; Frohly, Maxime; Willermain, François; Delporte, Christine; Motulsky, Elie

doi:10.3390/ijms24054509

Cited by 9 publications

(3 citation statements)

References 128 publications

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“…RPE cell migration is a crucial step in the formation of proliferative membranes in PVR. 27 , 28 During the PVR formation, RPE cells first detach from Bruch's membrane and migrate to the epi-retinal or sub-retinal spaces where they interact with other cellular components and the extracellular matrix to form the proliferative membrane. 7 , 8 In our study, both scratch assay and transwell assay demonstrated that silibinin significantly inhibited the migration of RPE cells.…”

Section: Discussionmentioning

confidence: 99%

Silibinin Prevents TGFβ-Induced EMT of RPE in Proliferative Vitreoretinopathy by Inhibiting Stat3 and Smad3 Phosphorylation

Ma,

Xie,

Gong

et al. 2023

Invest. Ophthalmol. Vis. Sci.

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Purpose The purpose of this study was to investigate the effects of silibinin on epithelial-mesenchymal transition (EMT) of retinal pigment epithelial (RPE) and proliferative vitreoretinopathy (PVR) formation, as well as its underlying molecular mechanism. Methods Cellular morphological change and EMT molecular markers were evaluated by using phase contrast imaging, qPCR, and Western blot (WB) to investigate the impact of silibinin on the EMT of ARPE-19 cells. Scratch assay and transwell assay were used to study the effect of silibinin on cell migration. An intravitreally injected RPE-induced rat PVR model was used to assess the effect of silibinin on PVR in vivo. RNA-seq was applied to study the molecular mechanism of silibinin-mediated PVR prevention. Results Silibinin inhibited TGFβ1-induced EMT and migration of RPE in a dose-dependent manner in vitro. Moreover, silibinin prevented proliferative membrane formation in an intravitreal injected RPE-induced rat PVR model. In line with these findings, RNA-seq revealed a global suppression of TGFβ1-induced EMT and migration-related genes by silibinin in RPEs. Mechanistically, silibinin reduced TGFβ1-induced phosphorylation levels of Smad3 and Stat3, and Smad3 nuclear translocation in RPE. Conclusions Silibinin inhibits the EMT of RPE cells in vitro and prevents the formation of PVR membranes in vivo. Mechanistically, silibinin inhibits Smad3 phosphorylation and suppresses Smad3 nuclear translocation through the inhibition of Stat3 phosphorylation. These findings suggest that silibinin may serve as a potential treatment for PVR.

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Section: Discussionmentioning

confidence: 99%

Silibinin Prevents TGFβ-Induced EMT of RPE in Proliferative Vitreoretinopathy by Inhibiting Stat3 and Smad3 Phosphorylation

Ma,

Xie,

Gong

et al. 2023

Invest. Ophthalmol. Vis. Sci.

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“…Neovascular age-related macular degeneration (nAMD) also develops membranes, similar to PVDs’ membranes, but the trigger for this pathology mainly consists of inflammation or oxidative stress. 1 …”

Section: Introductionmentioning

confidence: 99%

“…Neovascular age-related macular degeneration (nAMD) also develops membranes, similar to PVDs' membranes, but the trigger for this pathology mainly consists of inflammation or oxidative stress. 1 It has already been proven that retinal pigment epithelial (RPE) cells are deeply involved in PVDs' development, stimulated by cytokines or growth factors released as a consequence of the trigger events. 2 In particular, RPE cells, that in healthy condition are subjected to contact inhibitory effect and have a very low turnover rate, under a stimulatory event, for example retinal detachment in PVR, improve their proliferative and migratory properties, and undergo morphological changes, losing their cellular polarity and assuming a fibroblastoid appearance, leading to the so-called epithelial-mesenchymal transition (EMT).…”

Section: Introductionmentioning

confidence: 99%

Microfluidic Fabricated Liposomes for Nutlin-3a Ocular Delivery as Potential Candidate for Proliferative Vitreoretinal Diseases Treatment

Esposito,

Pozza,

Contado

et al. 2024

IJN

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Purpose Proliferative vitreoretinal diseases (PVDs) represent a heterogeneous group of pathologies characterized by the presence of retinal proliferative membranes, in whose development retinal pigment epithelium (RPE) is deeply involved. As the only effective treatment for PVDs at present is surgery, we aimed to investigate the potential therapeutic activity of Nutlin-3a, a small non-genotoxic inhibitor of the MDM2/p53 interaction, on ARPE-19 cell line and on human RPE primary cells, as in vitro models of RPE and, more importantly, to formulate and evaluate Nutlin-3a loaded liposomes designed for ophthalmic administration. Methods Liposomes were produced using an innovative approach by a microfluidic device under selection of different conditions. Liposome size distribution was evaluated by photon correlation spectroscopy and centrifugal field flow fractionation, while the liposome structure was studied by transmission electron microscopy and Fourier-transform infrared spectroscopy. The Nutlin-3a entrapment capacity was evaluated by ultrafiltration and HPLC. Nutlin-3a biological effectiveness as a solution or loaded in liposomes was evaluated by viability, proliferation, apoptosis and migration assays and by morphological analysis. Results The microfluidic formulative study enabled the selection of liposomes composed of phosphatidylcholine (PC) 5.4 or 8.2 mg/mL and 10% ethanol, characterized by roundish vesicular structures with 150–250 nm mean diameters. Particularly, liposomes based on the lower PC concentration were characterized by higher stability. Nutlin-3a was effectively encapsulated in liposomes and was able to induce a significant reduction of viability and migration in RPE cell models. Conclusion Our results lay the basis for a possible use of liposomes for the ocular delivery of Nutlin-3a.

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Critical role of apoptosis in MeCP2‐mediated epithelial–mesenchymal transition of ARPE‐19 cells

Zhang,

Wu,

Guo

et al. 2024

Journal Cellular Physiology

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Proliferative vitreoretinopathy (PVR) is a complex disease that significantly contributes to recurrent retinal detachment. Its development is notably affected by epithelial–mesenchymal transition (EMT), where apoptosis plays a crucial role as a regulator of EMT. However, the function of MeCP2 in governing apoptosis and EMT in retinal pigment epithelial (RPE) cells and its implications for PVR development have remained inadequately understood. Thus, we investigated the impact of MeCP2 on proliferation, migration, apoptosis and EMT in ARPE‐19 cells to provide a fresh perspective on the etiology of PVR. The morphological changes in ARPE‐19 cells induced by recombinant human MeCP2 protein and MeCP2 knockdown were observed. Wound healing assay were performed to verify the effects of recombinant human MeCP2 protein and MeCP2 knockdown on ARPE‐19 cell migration. Furthermore, cell proliferation was assessed using the CCK‐8 assay and flow cytometry. Western blot analysis, quantitative reverse transcriptase‐polymerase chain reaction (qRT‐PCR), and immunofluorescence analysis were conducted to measure the protein levels associated with apoptosis, cell cycle and EMT. Western blot analysis and immunofluorescence assays confirmed that MeCP2 promoted EMT formation in ARPE‐19 cells. The CCK‐8 assay revealed that MeCP2 treatment enhanced the proliferation of ARPE‐19 cells, whereas MeCP2 knockdown inhibited ARPE‐19 cell proliferation. Treatment with recombinant human MeCP2 protein and MeCP2 knockdown altered the morphology of ARPE‐19 cells. Wound healing assay demonstrated that MeCP2 knockdown inhibited ARPE‐19 cell migration, and MeCP2 treatment promoted ARPE‐19 cell migration. MeCP2 knockdown induced a G0/G1 phase block, inhibiting cell growth, and qRT‐PCR data indicated reduced expression of cell cycle‐related genes. Increased apoptosis was observed after MeCP2 knockdown in ARPE‐19 cells. Overall, MeCP2 treatment stimulates cell proliferation, migration and EMT formation; conversely, MeCP2 knockdown inhibits EMT, cell proliferation, migration and cell cycle G1/S phase transition, and induces apoptosis.

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Experimental Models to Study Epithelial-Mesenchymal Transition in Proliferative Vitreoretinopathy

Cited by 9 publications

References 128 publications

Silibinin Prevents TGFβ-Induced EMT of RPE in Proliferative Vitreoretinopathy by Inhibiting Stat3 and Smad3 Phosphorylation

Silibinin Prevents TGFβ-Induced EMT of RPE in Proliferative Vitreoretinopathy by Inhibiting Stat3 and Smad3 Phosphorylation

Microfluidic Fabricated Liposomes for Nutlin-3a Ocular Delivery as Potential Candidate for Proliferative Vitreoretinal Diseases Treatment

Critical role of apoptosis in MeCP2‐mediated epithelial–mesenchymal transition of ARPE‐19 cells

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