Characterization and Validation of In Vitro and In Vivo Models to Investigate TNF-α-Induced Inflammation in Retinal Diseases

Abstract

Purpose Inflammation is implicated in the etiology of diverse retinopathies including uveitis, age-related macular degeneration or diabetic retinopathy. Tumor necrosis factor alpha (TNF-α) is a well-known proinflammatory cytokine that is described as a biomarker for inflammation in diverse retinopathies and therefore emerged as an interesting target to treat inflammation in the eye by neutralizing anti-TNF-α antibodies. Methods Recently, we have demonstrated that Adeno-associated virus (AAV)–mediated expression of human TNF-α in the murine eye induces retinal inflammation including vasculitis and fibrosis, thereby mimicking human disease-relevant pathologies. In a proof-of-mechanism study, we now tested whether AAV-TNF-α induced pathologies can be reversed by neutralizing TNF-α antibody treatment. Results Strikingly, a single intravitreal injection of the TNF-α antibody golimumab reduced AAV-TNF-α–induced retinal inflammation and retinal thickening. Furthermore, AAV-TNF-α–mediated impaired retinal function was partially rescued by golimumab as revealed by electroretinography recordings. Finally, to study TNF-α-induced vasculitis in human in vitro cell culture assays, we established a monocyte-to-endothelium adhesion co-culture system. Indeed, also in vitro TNF-α induced monocyte adhesion to human retinal endothelial cells, which was prevented by golimumab. Conclusions Overall, our study describes valuable in vitro and in vivo approaches to study the function of TNF-α in retinal inflammation and demonstrated a preclinical proof-of-mechanism treatment with golimumab. Translational Relevance The AAV-based model expressing human TNF-α allows us to investigate TNF-α–driven pathologies supporting research in mechanisms of retinal inflammation.

Transcriptome analysis of AAV-induced retinopathy models expressing human VEGF, TNF-α, and IL-6 in murine eyes

“…4b). To validate our results, we measured C3 expression by ELISA in an independent mouse cohort 57 and verified the upregulation of also C3 protein at 6 weeks after AAV-TNF-α treatment (Fig. 4c).…”

“…Tissue lysis and ELISA. Eyes for validation of C3 expression were generated from an independent mouse cohort that is published elsewhere 57 . In brief, 2 weeks after IVT injection of AAV-TNF-α, 100 µg golimumab (Simponi) were injected intravitreally.…”

IL-23 drives uveitis by acting on a novel population of tissue-resident entheseal T cells

“…Following injection, constitutive secretion of IL-23 led to rapid onset of inflammation, followed by a chronic and persistent disease phenotype not observed or attributed to ShH10_GFP response. Use of AAV to model, probe and evaluate therapeutics for human inflammatory ocular disease is an expanding area [81, 82], and in the current context provides a tool to interrogate pathogenicity of IL-23 alone in a disease that is accom-panied by elevated concentrations of serum and AqH IL-23 [39, 41, 42, 83].…”

Protective mechanism of Scutellarin against pyroptosis network in diabetic retinopathy

“…A study indicated that [54] the expression of TNF-α mRNA was signi cantly increased in diabetic rats induced by a single intraperitoneal injection of 60 mg/kg STZ. A research has demonstrated that [55] Adeno-associated virus (AAV)-mediated expression of human TNF-α in the murine eye induces retinal in ammation. More importantly, TNF-α released from the activated microglia induced BRB damage, which in turn leaded to in ammation of the retinal [56].…”