Abstract
Purpose :
Age-related macular degeneration (AMD) is a multifactorial disease that leads to gradual vision loss and is correlated with a combination of environmental factors including age and lifestyle, and genetic predisposition where many of the targets associated with AMD risk are complement related. Various models of atrophic AMD have been produced in the literature, but none have fully assessed complement’s role in AMD progression. We therefore sought to develop our own in-house in vitro models of complement induced stress.
Methods :
Using an RPE-centric approach, we differentiated iPSCs derived from intermediate AMD and geographic atrophy patients into RPE cells and stimulated the cells with normal human serum. We evaluated cell morphology, barrier function via transepithelial resistance (TEER), epithelial to mesenchymal transition and crucial cellular responses.
Results :
We observe a reduction in TEER by approximately 52.43% (449.85 Ωcm2; p = 0.0138) after 1 hour of stimulation and eventual morphological changes by 24 hours. With complement induction, we noted a robust increase of C3a and C5a in the apical supernatant by 24 hours post stimulation. In addition, an increase of various cytokines including IL-1β, IL-6, IL-8, and IL-13 (p < 0.0001) by 48 hours. There was no significant increase of cytokine or complement factor production in the basolateral supernatant.
Conclusions :
With our models, we note various changes in cellular morphology, function, and responses. Pathway analysis identified several downstream effects of complement-induced stress and possible targets of interest. Using our approach, we hope to deconvolute the initiation of AMD progression.
This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.