Differential effects of risuteganib and bevacizumab on AMD cybrid cells
| Autor: | Marilyn Chwa, Zixuan Shao, Vicken H. Karageozian, Shari R. Atilano, M. Cristina Kenney, Kevin Schneider, Hampar L. Karageozian, John Y Park |
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| Rok vydání: | 2021 |
| Předmět: |
Male
Vascular Endothelial Growth Factor A 0301 basic medicine Aging genetic structures Angiogenesis Angiogenesis Inhibitors Retinal Pigment Epithelium Neurodegenerative Medical Biochemistry and Metabolomics Eye Ophthalmology & Optometry medicine.disease_cause Macular Degeneration 0302 clinical medicine 80 and over 2.1 Biological and endogenous factors Risuteganib Aetiology Aged 80 and over Caspase 7 Membrane Potential Mitochondrial Caspase 3 Chemistry Intracellular Signaling Peptides and Proteins Sensory Systems Mitochondrial Bevacizumab Vascular endothelial growth factor A Female Blood Platelets Cybrids SOD2 Hybrid Cells Real-Time Polymerase Chain Reaction Membrane Potential DNA Mitochondrial Article Cell Line 03 medical and health sciences Cellular and Molecular Neuroscience Opthalmology and Optometry medicine Humans Eye Disease and Disorders of Vision Aged Age-related macular degeneration Neurosciences DNA eye diseases Oxidative Stress Ophthalmology 030104 developmental biology Gene Expression Regulation Mitochondrial biogenesis Apoptosis BCL2L13 Wet Macular Degeneration 030221 ophthalmology & optometry Cancer research sense organs Peptides Reactive Oxygen Species Oxidative stress |
| Zdroj: | Exp Eye Res |
| ISSN: | 0014-4835 |
| DOI: | 10.1016/j.exer.2020.108287 |
| Popis: | PurposeIntravitreal injections of anti-vascular endothelial growth factor (VEGF) treatments are currently used to treat wet age-related macular degeneration (AMD), diabetic retinopathy, and macular edema. Chronic, repetitive treatments with anti-VEGF may have unintended consequences beyond the inhibition of angiogenesis. Most recently, clinical trials have been conducted with risuteganib (RSG, Luminate®), which is anti-angiogenic and has neuroprotective and anti-inflammatory properties. Mitochondrial damage and dysfunction play a major role in development of AMD. Transmitochondrial cybrids are cell lines established by fusing human retinal pigment epithelial (RPE) cells that are Rho0 (lacking mtDNA) with platelets isolated from AMD subjects or age-matched normal subjects. Cybrid cell lines have identical nuclei but mitochondria from different subjects, enabling investigation of the functional consequences of damaged AMD mitochondria. The present study compares the responses of AMD cybrids treated with bevacizumab (Bmab, Avastin®) versus risuteganib (RSG, Luminate®).MethodsCybrids were created by fusing mtDNA depleted ARPE-19cells with platelets from AMD or age-matched normal patients. AMD (n=5) and normal (n=3) cybrids were treated for 48h with or without 1x clinical dose of 1.25 mg/50μl (25,000μg/ml) of Bmab or 1.0 mg/50μl (20,000μg/ml) of RSG. Cultures were analyzed for levels of cleaved caspase 3/7 and NucLight Rapid Red staining (IncuCyte® Live Cell Imager), mitochondrial membrane potential (ΔΨm, JC1 assay) or reactive oxygen species (ROS, H2DCFDA assay). Expression levels of genes related to the following pathways were analyzed with qRT-PCR: Apoptosis (BAX, BCL2L13, CASP-3, -7, -9); angiogenesis (VEGFA, HIF1α, PDGF); integrins (ITGB-1, -3, -5, ITGA-3, -5, -V); mitochondrial biogenesis (PGC1α, POLG); oxidative stress (SOD2, GPX3, NOX4); inflammation (IL-6, -18, -1β, IFN-β1); and signaling (P3KCA, PI3KR1). Statistical analyses were performed using GraphPad Prism software.ResultsThe untreated AMD cybrids had significantly higher levels of cleaved caspase 3/7 compared to the untreated normal cybrids. The Bmab-treated AMD cybrids showed elevated levels of cleaved caspase 3/7 compared to untreated AMD or RSG-treated AMD cybrids. The Bmab-treated cybrids had lower ΔΨm compared to untreated AMD or RSG-treated AMD cybrids. The ROS levels were not changed with Bmab or RSG treatment. Results showed that Bmab-treated cybrids had higher expression levels of inflammatory (IL-6, IL1-β), oxidative stress (NOX4) and angiogenesis (VEGFA) genes compared to untreated AMD, while RSG-treated cybrids had lower expression levels of apoptosis (BAX), angiogenesis (VEGFA) and integrin (ITGB1) genes.ConclusionsThese data suggest that the mechanism(s) of action of RSG, an integrin regulator, and Bmab, a recombinant monoclonal antibody, affect the AMD RPE cybrid cells differently, with the former having more anti-apoptosis properties, which may be desirable in treating degenerative ocular diseases. |
| Databáze: | OpenAIRE |
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