Page 33 - Read Online

Plössl et al. J Transl Genet Genom 2022;6:46-62 Journal of Translational
DOI: 10.20517/jtgg.2021.39
Genetics and Genomics

Original Article Open Access

In vitro modeling of the complex retinal condition
age-related macular degeneration

1,2
1
1,#
1,#
Karolina Plössl , Emily Webster , Christina Kiel , Felix Grassmann , Caroline Brandl 1,3,4 , Bernhard H. F.
Weber 1,5
1
Institute of Human Genetics, University of Regensburg, Regensburg 93053, Germany.
2
Institute of Medical Sciences, University of Aberdeen, King’s College, Aberdeen AB24 3FX, UK.
3
Department of Ophthalmology, University Hospital Regensburg, Regensburg 93053, Germany.
4
Department of Genetic Epidemiology, University of Regensburg, Regensburg 93053, Germany.
5
Institute of Clinical Human Genetics, University Hospital Regensburg, Regensburg 93053, Germany.
#
Authors contributed equally and considered joint first authors.
Correspondence to: Bernhard H. F. Weber, Institute of Human Genetics, University of Regensburg, Franz-Josef-Strauss Allee 11,
Regensburg 93053, Germany. E-mail: bweb@klinik.uni-regensburg.de
How to cite this article: Plössl K, Webster E, Kiel C, Grassmann F, Brandl C, Weber BHF. In vitro modeling of the complex retinal
condition age-related macular degeneration. J Transl Genet Genom 2022;6:46-62. https://dx.doi.org/10.20517/jtgg.2021.39
Received: 12 Aug 2021 First Decision: 9 Sep 2021 Revised: 8 Oct 2021 Accepted: 19 Oct 2021 Published: 7 Jan 2022
Academic Editor: Sanjay Gupta Copy Editor: Yue-Yue Zhang Production Editor: Yue-Yue Zhang

Abstract
Aim: To model a complex retinal disease such as age-related macular degeneration (AMD) in vitro, we aimed to
combine genetic and environmental risk factors in a retinal pigment epithelium (RPE) cell culture model generated
via induced pluripotent stem cells (iPSCs) from subjects with an extremely high and an extremely low genetic
disease risk. As an external stimulus, we chose defined oxidative stress conditions.
Methods: Patients were genotyped for known AMD-associated genetic variants and their individual genetic risk
score (GRS) was calculated defining individual iPSC-RPE cell lines which reflect the extreme ends of the genetic
risk for AMD. Sodium iodate (NaIO , SI) was used to induce oxidative stress and cellular responses were followed
3
by analyzing nuclear factor erythroid 2-related factor 2 (NRF2) pathway activation by mRNA and protein
expression.

Results: We present a collection of eight iPSC-RPE cell lines, with four each harboring an extreme low or an
extreme high GRS for AMD. RPE identity was verified structurally and functionally. We found that 24 and 72 h of SI
treatment induced a significant upregulation of NRF2 response genes HMOX1 and NQO1, without showing cytotoxic
effects or negatively influencing RPE cell integrity. High- vs. low-risk cell lines revealed similar first line defenses in

© The Author(s) 2022. Open Access This article is licensed under a Creative Commons Attribution 4.0
International License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, sharing,
adaptation, distribution and reproduction in any medium or format, for any purpose, even commercially, as
long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and
indicate if changes were made.

www.jtggjournal.com