We have utilized an existing repository of AMD patient clinical data (AREDS2) that includes 5-10 years of extensive clinical imaging and medical history, combined with genomic data to generate a comprehensive set of iPSC-derived RPE for disease pathway analysis and genotype-phenotype association. In vitro analysis of iPSC-RPE can be used to link RPE signaling pathways to AMD risk alleles and thus to various stages of AMD. This allows making a direct link between RPE signaling pathways, AMD risk alleles, and AMD phenotype. To help accomplish this goal we have established a collaboration with the New York Stem Cell Foundation (NYSCF) who have generated over 60 AMD iPSC-derived RPE and, in some few cases, twin controls and provide storage and distribution of these cell lines to the entire scientific community.
RPE shape, pigmentation, and autofluorescent changes using live-imaging (software developed at NEI IRP). These findings will be correlated to in vivo changes in RPE shape and pigmentation and analyzed longitudinally in patients using adaptive optics.
Formation of drusen-like particles and their polarized secretion by iPSC-derived RPE cells (Pilgrim et al., 2017, IOVS) to determine a direct relationship between AMD genetics and drusen or pseudoreticular drusen formation.
Specific intracellular autophagic pathways using fluorescent reporters (Barmada et al., 2014) to identify potentially druggable targets to slow down disease initiation or progression (The HD iPSC Consortium, Nat. Neuroscience, epub March 2017 – Analysis of iPSC-neurons derived from Huntington disease patients demonstrates that mutant huntingtin impairs neurodevelopmental pathways that could disrupt synaptic homeostasis and increase vulnerability to the pathologic consequence of expanded polyglutamine repeats over time. Some of these phenotypes could be reversed by the treatment of isoxazole-9).
The polarized secretion by human RPE inflammatory cytokines - IL6, Il1-beta, IL18, and VEGF as well as the expression of miRNAs (eg miRs-155, 204) that have recently been shown to activate RPE inflammasome activity and AMD pathophysiology (Shi et al., 2008, IOVS; Wang et al., 2010 Hum. Gen.; Ambati et al., 2013 Nat. Rev. Immunology; Campbell, 2014 Adv. Exp. Med. Biol; Celkova, et al., 2015 J. Clin. Med; Berber et al., 2017 Mol. Diagn. Ther.).