June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
iPSC-RPE with CFH Y402H risk variant show higher disease propensity compared to the non-risk CFH variant
Author Affiliations & Notes
  • Ruchi Sharma
    National Eye Institute, Bethesda, Maryland, United States
  • Malika Nimmagadda
    University of Wisconsin System, Madison, Wisconsin, United States
  • Davide Ortolan
    National Eye Institute, Bethesda, Maryland, United States
  • Kelcy Klein
    National Eye Institute, Bethesda, Maryland, United States
  • Jair Montford
    National Eye Institute, Bethesda, Maryland, United States
  • Kapil Bharti
    National Eye Institute, Bethesda, Maryland, United States
  • Footnotes
    Commercial Relationships   Ruchi Sharma None; Malika Nimmagadda None; Davide Ortolan None; Kelcy Klein None; Jair Montford None; Kapil Bharti None
  • Footnotes
    Support  none
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 2431 – F0375. doi:
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      Ruchi Sharma, Malika Nimmagadda, Davide Ortolan, Kelcy Klein, Jair Montford, Kapil Bharti; iPSC-RPE with CFH Y402H risk variant show higher disease propensity compared to the non-risk CFH variant. Invest. Ophthalmol. Vis. Sci. 2022;63(7):2431 – F0375.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : The alternate complement pathway and age-related macular degeneration (AMD) have been long connected via genetic association studies. The complement factor H (CFH) gene has one of the highest odds ratio for the disease propensity, and dysregulated alternate complement pathway is thought to accelerate the disease phenotype. RPE cells with the highest CFH expression are the likely initiation site for complement attack. Using iPSC-RPE derived from patients with risk(Y402H) and non-risk(H402H) CFH alleles, we aim to decipher the role of CFH in disease propensity under a complement attack and determine genotype to phenotype interaction.

Methods : Four iPSC lines from CFH risk Y402H and three from non-risk H402H variants were differentiated into RPE and confirmed for functionality and maturity. To check if CFH risk-allele containing iPSC-RPE show signs of vulnerability to complement attack, we tested secreted CFH, C3a, and C5a levels, phagocytic ability, monolayer transepithelial resistance (TER) with and without complement serum. In addition, cells were stained with sub-RPE deposit marker-APOE, lipid droplets with Nile red, and BODIPY and complement lytic attack complex to check the AMD phenotype.

Results : CFH risk variant iPSC-RPE showed lower outer segment digestion capacity, and lower TER. CFH high-risk cells also had significantly higher sub-RPE and lipid deposits than CFH non-risk variant iPSC-RPE, suggesting disease vulnerability. The lower CFH secretion and higher C3a and C5a levels in CFH risk variants likely is the downstream mechanism making these RPE cells susceptible to AMD.

Conclusions : iPSC-RPE from the CFH risk variant recapitulated the AMD phenotype at basal levels and under complement attack- creating AMD model in a dish. This study provides a tool to decipher genotype interaction with macular degeneration phenotype and other retinal dystrophies. In addition, this platform can be used to further study interactions between different risk alleles such as ARMS2/HTRA1 and to perform drug screens to discover potential therapeutic agents.

This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.

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