Unraveling the web of nAMD: A transcriptomic analysis of age-related macular degeneration-associated cytokines in primary human retinal endothelial cells.

Neovascular age-related macular degeneration (nAMD) is a complex, blinding retinal disease underpinned by a dynamic and dysregulated interaction among pathogenic cytokines. The present study investigates the impact of six nAMD-associated cytokines, namely transforming growth factor-beta 1 and 2 (TGF-β1/2), tumour necrosis factor-alpha (TNF-α), thrombin, interleukin-6 (IL-6) and vascular endothelial growth factor (VEGF), on the transcriptome of primary human retinal endothelial cells (HRECs), both individually and as a collective 'cocktail'. HRECS were treated for 24 hours before being harvested for bulk RNA-seq. In performing this study, we have described the different transcriptomic signatures of each cytokine in driving nAMD-associated inflammatory, angiogenic and fibrotic pathways when presented in isolation to primary HRECs. Additionally, this study dissects a novel 'cocktail’ effect in the collective group characterised by further differential expression of genes, some of which were not induced by any of the individual cytokines. We explore the translatability of our study as an in vitro model of nAMD in endothelial cells by mapping the identified differential expression patterns to marker gene sets identified in previously performed scRNASeq of nAMD-affected human donor eyes.