Prospects and limitations of cumate-inducible lentivirus as a tool for investigating VEGF-A-mediated pathology in diabetic retinopathy

• Published in: Scientific reports Vol. 14; no. 1; pp. 14325 - 12
• Main Authors: Lelyte, Inesa, Rao, Vidhya R., Kalesnykas, Giedrius, Ragauskas, Symantas, Kaja, Simon, Ahmed, Zubair
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 21.06.2024; Nature Publishing Group; Nature Portfolio
• Subjects: 631/154; 631/208; 631/378; 631/80; Animal models; Animals; Cell growth; Cell Line; Cell Movement; Cell Proliferation; Cell Survival; Cumate-inducible lentivirus; Diabetes; Diabetes mellitus; Diabetic retinopathy; Diabetic Retinopathy - metabolism; Diabetic Retinopathy - pathology; Disease Models, Animal; Feasibility studies; Gene expression; Genetic Vectors - administration & dosage; Genetic Vectors - genetics; Gliosis; Hemorrhage; Humanities and Social Sciences; Humans; Immunohistochemistry; Intravitreal Injections; Lentivirus - genetics; Male; mRNA; multidisciplinary; Pathology; Permeability; Protein folding; Rats; Rats, Wistar; Retina; Retinal pigment epithelium; Retinal Pigment Epithelium - metabolism; Retinal Pigment Epithelium - pathology; Retinal toxicity; Retinopathy; Science; Science (multidisciplinary); Tomography, Optical Coherence; Toxicity; Vascular endothelial growth factor; Vascular Endothelial Growth Factor A - genetics; Vascular Endothelial Growth Factor A - metabolism; Vascularization; Diabetic retinopathy; Retinal toxicity; Gene expression; Vascular endothelial growth factor; Cumate-inducible lentivirus
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-024-63590-y

Diabetic retinopathy (DR) is a multifactorial disease displaying vascular-associated pathologies, including vascular leakage and neovascularization, ultimately leading to visual impairment. However, animal models accurately reflecting these pathologies are lacking. Vascular endothelial growth factor A (VEGF-A) is an important factor in the development of micro- and macro-vascular pathology in DR. In this study, we evaluated the feasibility of using a cumate-inducible lentivirus (LV) mediated expression of vegf-a to understand DR pathology in vitro and in vivo. Retinal pigment epithelial cells (ARPE-19) were transduced with cumate-inducible LV expressing vegf-a , with subsequent analysis of vegf-a expression and its impact on cell proliferation, viability, motility, and permeability. Cumate tolerability in adult Wistar rat eyes was assessed as an initial step towards a potential DR animal model development, by administering cumate via intravitreal injections (IVT) and evaluating consequent effects by spectral domain optical coherence tomography (SD-OCT), flash electroretinography (fERG), ophthalmic examination (OE), and immunohistochemistry. Transduction of ARPE-19 cells with cumate-inducible LV resulted in ~ 2.5-fold increase in vegf-a mRNA and ~ threefold increase in VEGF-A protein secretion. Transduced cells displayed enhanced cell proliferation, viability, permeability, and migration in tube-like structures. However, IVT cumate injections led to apparent retinal toxicity, manifesting as retinal layer abnormalities, haemorrhage, vitreous opacities, and significant reductions in a- and b-wave amplitudes, along with increased microglial activation and reactive gliosis. In summary, while cumate-inducible LV-mediated vegf-a expression is valuable for in vitro mechanistic studies in cellular drug discovery, its use is not a feasible approach to model DR in in vivo studies due to cumate-induced retinal toxicity.