Bevacizumab Induces Upregulation of Keratin 3 and VEGFA in Human Limbal Epithelial Cells in Vitro

Abstract

Topical application of vascular endothelial growth factor A (VEGFA) inhibitors including Bevacizumab is used for antiangiogenic therapy at the ocular surface. While clinical studies have suggested that this approach is well-tolerated, the effect of the drug on limbal epithelial stem cells has not been studied. In this study, the effect of Bevacizumab on phenotype and functionality of putative limbal epithelial stem cells (SC) was investigated. The effect of Bevacizumab on human limbal epithelial cells was assessed in terms of metabolic activity and scratch wound closure. The different treatment groups featured no difference in proliferation and colony forming efficiency (CFE) of limbal epithelial cells or their putative SC marker expression. A significant delay in scratch closure of all the Bevacizumab-treated groups was detected at 4 h. RNA and protein quantification indicated a dose-responsive increase of keratin 3. VEGFA RNA expression also increased while VEGFC and D as well as VEGFR1, 2 and 3 were unchanged. This study highlights previously unknown effects of Bevacizumab on cultured putative limbal epithelial SC: a dose-related increase of keratin 3, an increase in VEGFA as well as a delay in scratch wound closure. These in vitro data should be considered when using Bevacizumab in the context of limbal epithelial SC transplantation.

Targeted dexamethasone nano-prodrug for corneal neovascularization management

“…VEGF is a critical pro-angiogenic molecule that binds to its receptor to increase vascular permeability and promote angiogenesis by stimulating endothelial cells sprouting and proliferation [ 29 ]. In several VEGF isoforms, VEGF-A, VEGF-C, and VEGF-D are important in CNV through inducing the transcription of various angiogenesis-associated genes [ 30 , 31 ]. MMPs, proteolytic enzymes that degrade extracellular matrix, are involved in angiogenesis inhibition and interference of vascular maturation steps.…”

Profile of biological characterizations and clinical application of corneal stem/progenitor cells

“…Moreover, cytokeratin K3 and K12, as specific markers of the corneal epithelial phenotype, were not expressed in limbal basal epithelial cells. Therefore, many studies have also confirmed the low differentiation phenotype of LSCs[ 20 , 21 ]. Many studies have shown that some materials or cells [such as Frizzled 7, HC-HA/PTX3 and human limbal melanocytes (hLM)] can maintain the low differentiation state of LSCs[ 22 ] (Niche regulation of limbal epithelial stem cells: HC-HA/PTX3 as a surrogate matrix niche) (Limbal melanocytes support limbal epithelial stem cells in 2D and 3D microenvironments).…”

Inhibitory effect of anti-Scg3 on corneal neovascularization: a preliminary study

“…laser photocoagulation and photodynamic therapy only could cause temporary occlusion and withdrawal to relieve symptoms [ 11 ]. Notara et al found that the high concentration of bevacizumab would downregulate the integrin α and β, and which interferes with the adhesion mechanism of cell migration and affects corneal epithelial healing [ 12 ], resulting in thinning of the corneal stroma [ 13 ]. Yin et al found the CNV progressed in the discontinuation of bevacizumab sometimes [ 14 ].…”