Investigating the Effect of Aflibercept and Dexamethasone on the Expression Level of Genes Associated with Epithelial-Mesenchymal Transition (EMT) in Retinal Pigment Epithelial (RPE) at Transcriptional Level
Hajar Farvardin1 *, Mohammad Hossein Nowroozzadeh1 , Fatemeh Sanie-Jahromi1 , Ali Nabavizadegan1
- Poostchi Ophthalmology Research Center, Department of Ophthalmology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran
Abstract: Background: Proliferative vitreoretinopathy (PVR) is the major cause of surgical failure in retinal detachment repair and epithelial-mesenchymal transition (EMT) of RPE cells plays a pivotal role in the pathophysiology of PVR. The main purpose of this study is to investigate the impact of aflibercept, alone and combined with dexamethasone on the genes involved in the epithelial-mesenchymal transition of RPE cells.
Methods: Methods: Human cadaver eyeballs from a male organ donor underwent processing for RPE cell culture in a sterile lab environment. RPE cells were further divided into 5 Groups: Group 1 received aflibercept, Group 2 received bevacizumab, Group 3 received dexamethasone, Group 4 received aflibercept + dexamethasone, and Group 5 served as the control group. We examined the expression levels of 6 different EMT-related genes (α-SMA, ZEB1, TGF-β, CD90, β-catenin, and Snail) through RNA extraction, cDNA synthesis, and real-time PCR following a 24-hour treatment. Each test was repeated at least three times to ensure reliable outcomes.
Results: Results: The expression pattern of EMT-related genes in RPE cells receiving anti-VEGF agents (Group 1 and 2) showed similarities, especially in the significant reduction of ?-SMA and ZEB1 genes. The expression levels of TFG-? and CD90 were significantly lower in cells treated with dexamethasone alone and combined with aflibercept. The gene expression profile of Group 4 (dexamethasone plus aflibercept) was more similar to Group 3 rather than the first two groups. We also observed a significant increase in the expression of some EMT genes (?-catenin and Snail) in Groups 3 and 4, a pattern not seen in the absence of dexamethasone.
Conclusion: Conclusions: The gene expression profiles observed in this study revealed promising patterns in cells treated with aflibercept, bevacizumab, and dexamethasone, both individually and in combination. Investigating the impact of anti-VEGF agents on EMT-associated genes in RPE cells provides valuable insights into the new pharmacologic treatment designs for proliferative vitreoretinopathy.
