In vitro response and gene expression of human retinal Müller cells treated with different anti-VEGF drugs

Cáceres-del-Carpio, Javier; Moustafa, M. Tarek; Toledo-Corral, Jaime; Hamid, Mohamed Abdel; Atilano, Shari R.; Schneider, Kevin; Fukuhara, Paula Sakemi; Costa, Rodrigo Donato; Norman, Jon Lucas; Malik, Deepika; Chwa, Marilyn; Boyer, David S.; Limb, G. Astrid; Kenney, M. Cristina; Kuppermann, Baruch D.

doi:10.1016/j.exer.2019.107903

Cited by 8 publications

(4 citation statements)

References 47 publications

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“…Indeed, as shown by our immunofluorescence data, they seem to be the only retinal cell type, together with endothelial cells, displaying evident VEGFR2 expression, and both MIO-M1 cells and Müller cells in retinal explants greatly increase VEGFR2 expression in response to exo-VEGF. Müller cells are the main source of VEGF in the retina and they release VEGF in pathologic conditions such as DR [ 42 , 43 , 44 ]. In addition, VEGF released by Müller cells is protective for the Müller cells themselves and for retinal neurons [ 21 , 22 ].…”

Section: Discussionmentioning

confidence: 99%

Oxidative Stress Induces a VEGF Autocrine Loop in the Retina: Relevance for Diabetic Retinopathy

Rossino

Lulli

Amato

et al. 2020

Cells

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Background: Oxidative stress (OS) plays a central role in diabetic retinopathy (DR), triggering expression and release of vascular endothelial growth factor (VEGF), the increase of which leads to deleterious vascular changes. We tested the hypothesis that OS-stimulated VEGF induces its own expression with an autocrine mechanism. Methods: MIO-M1 cells and ex vivo mouse retinal explants were treated with OS, with exogenous VEGF or with conditioned media (CM) from OS-stressed cultures. Results: Both in MIO-M1 cells and in retinal explants, OS or exogenous VEGF induced a significant increase of VEGF mRNA, which was abolished by VEGF receptor 2 (VEGFR-2) inhibition. OS also caused VEGF release. In MIO-M1 cells, CM induced VEGF expression, which was abolished by a VEGFR-2 inhibitor. Moreover, the OS-induced increase of VEGF mRNA was abolished by a nuclear factor erythroid 2-related factor 2 (Nrf2) blocker, while the effect of exo-VEGF resulted Nrf2-independent. Finally, both the exo-VEGF- and the OS-induced increase of VEGF expression were blocked by a hypoxia-inducible factor-1 inhibitor. Conclusions: These results are consistent with the existence of a retinal VEGF autocrine loop triggered by OS. This mechanism may significantly contribute to the maintenance of elevated VEGF levels and therefore it may be of central importance for the onset and development of DR.

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Section: Discussionmentioning

confidence: 99%

Oxidative Stress Induces a VEGF Autocrine Loop in the Retina: Relevance for Diabetic Retinopathy

Rossino

Lulli

Amato

et al. 2020

Cells

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“…Regarding the underlying mechanism, the promotion of gliosis of activated glial cells into fibrosis is induced by cytokines or profibrotic factors in the microenvironment [44][45] . Anti-VEGF agents cause structural protein abnormalities in cultured Müller cells, with decreased levels of GFAP and vimentin, probably due to the VEGF family receptors expressed in Müller cells [46][47] . Another way in which IVR can promote GMT is the neutralizing influence of VEGF on the inductive effects of released profibrotic factors, such as transforming growth factor (TGF)-β [48][49] , which is supported by previous studies showing that α-SMA or fibronectin in myofibroblasts depends on profibrotic cytokines, and that GMT in diabetic fibrovascular proliferation is driven by autoinduction of TGF-β in Müller glial cells [15,50] .…”

Section: Discussionmentioning

confidence: 99%

Effect of intravitreal ranibizumab on fibrovascular membranes in patients with proliferative diabetic retinopathy

Liang¹,

Li²,

Yang³

et al. 2022

Int J Ophthalmol

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AIM: To assess the effects of intravitreal ranibizumab (IVR) on angiogenesis and glial activity of the fibrovascular membrane (FVM) in patients with proliferative diabetic retinopathy (PDR). METHODS: Forty-two eyes from 42 patients with PDR requiring vitrectomy were included and divided into two groups: control group (n=16) did not receive IVR, while IVR group (n=26) underwent IVR 5d before vitrectomy. FVM specimens were collected by the same surgeon during the interventions. Histopathological morphology was examined by hematoxylin-eosin (H-E) staining and cell densities in the FVM was assessed. Microvessels were outlined by immunohistochemical staining of CD31 and microvessel density (MVD) assessed as an index of FVM angiogenesis. Dual-color immunofluorescence staining, and confocal microscopy was used to detect co-localization and relative expression levels of glial fibrillary acidic protein (GFAP) and α-smooth muscle actin (α-SMA) as markers of glial-mesenchymal transition (GMT). The GMT index (GI; ratio of relative GFAP/α-SMA expression) was used to semi-quantify the degree of GMT or glial activity of FVMs. RESULTS: H-E staining showed similar vascularization in both groups, with microvessels and scattered stromal cells in the matrix. Infiltrated cell densities did not differ significantly between the two groups (P>0.05). The MVD of the IVR group (130.62±15.46/mm2) was significantly lower than that of the controls (142.25±19.16/mm2, P<0.05). In both groups, all sections were strongly immunostained for GFAP and α-SMA. The Pearson's correlation coefficients (PCC) of intensity of automated pixel count of two markers indicated GFAP and α-SMA co-stained well and GMT participated in the remolding of FVMs in PDR. The mean relative GFAP expression in the IVR group was significantly lower, whereas that of α-SMA was significantly higher than in controls (P<0.05). GI in the IVR group (1.10±0.10) was significantly lower than in the controls (1.21±0.12, P<0.05). CONCLUSION: IVR can reduce angiogenesis, glial activity of FVM and promote glial-fibrotic transformation by reducing MVD and promoting GMT but does not decrease the cell density in patients with PDR.

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“…Previous observations had shown the presence of the VEGFR2 protein in Müller cells of rat and murine retina, VEGF neutralization or Vegfr2 disruption under diabetic conditions leading to Müller cell apoptosis in the two animal models, respectively [ 51 , 52 ]. On the other hand, treatment of MIO-M1 cells with anti-VEGF agents have led to contrasting results with modest or no effect on cell viability/apoptosis [ 53 , 54 ]. Further experiments are required to fully elucidate the role of the VEGF/VEGFR2 system in Müller cells.…”

Section: Discussionmentioning

confidence: 99%

VEGF-Independent Activation of Müller Cells by the Vitreous from Proliferative Diabetic Retinopathy Patients

Rezzola

Guerra

Chandran

et al. 2021

IJMS

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Proliferative diabetic retinopathy (PDR), a major complication of diabetes mellitus, results from an inflammation-sustained interplay among endothelial cells, neurons, and glia. Even though anti-vascular endothelial growth factor (VEGF) interventions represent the therapeutic option for PDR, they are only partially efficacious. In PDR, Müller cells undergo reactive gliosis, produce inflammatory cytokines/chemokines, and contribute to scar formation and retinal neovascularization. However, the impact of anti-VEGF interventions on Müller cell activation has not been fully elucidated. Here, we show that treatment of MIO-M1 Müller cells with vitreous obtained from PDR patients stimulates cell proliferation and motility, and activates various intracellular signaling pathways. This leads to cytokine/chemokine upregulation, a response that was not mimicked by treatment with recombinant VEGF nor inhibited by the anti-VEGF drug ranibizumab. In contrast, fibroblast growth factor-2 (FGF2) induced a significant overexpression of various cytokines/chemokines in MIO-M1 cells. In addition, the FGF receptor tyrosine kinase inhibitor BGJ398, the pan-FGF trap NSC12, the heparin-binding protein antagonist N-tert-butyloxycarbonyl-Phe-Leu-Phe-Leu-Phe Boc2, and the anti-inflammatory hydrocortisone all inhibited Müller cell activation mediated by PDR vitreous. These findings point to a role for various modulators beside VEGF in Müller cell activation and pave the way to the search for novel therapeutic strategies in PDR.

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In vitro response and gene expression of human retinal Müller cells treated with different anti-VEGF drugs

Cited by 8 publications

References 47 publications

Oxidative Stress Induces a VEGF Autocrine Loop in the Retina: Relevance for Diabetic Retinopathy

Oxidative Stress Induces a VEGF Autocrine Loop in the Retina: Relevance for Diabetic Retinopathy

Effect of intravitreal ranibizumab on fibrovascular membranes in patients with proliferative diabetic retinopathy

VEGF-Independent Activation of Müller Cells by the Vitreous from Proliferative Diabetic Retinopathy Patients

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