June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
Activation of autophagy reverses progressive and deleterious protein aggregation in PRPF31 patient induced pluripotent stem cells (iPSCs)-derived retinal pigment epithelium (RPE) cells.
Author Affiliations & Notes
  • Maria Georgiou
    Biosciences Institute, Newcastle upon Tyne, Tyne and Wear, United Kingdom
  • Chunbo Yang
    Biosciences Institute, Newcastle upon Tyne, Tyne and Wear, United Kingdom
  • Robert Atkinson
    Biosciences Institute, Newcastle upon Tyne, Tyne and Wear, United Kingdom
  • Kuan-Ting Pan
    Max-Planck Institute of Biophysical Chemistry, Göttingen, Germany, Göttingen,, Germany
  • Adriana Buskin
    Biosciences Institute, Newcastle upon Tyne, Tyne and Wear, United Kingdom
  • Marina Moya Molina
    Biosciences Institute, Newcastle upon Tyne, Tyne and Wear, United Kingdom
  • Joseph Collin
    Biosciences Institute, Newcastle upon Tyne, Tyne and Wear, United Kingdom
  • Sebastian E. J. Ludwig
    Max-Planck Institute of Biophysical Chemistry, Göttingen, Germany, Göttingen,, Germany
  • Sushma Nagaraja-Grellscheid
    University of Bergen, Norway, Department of Biological Sciences, Norway
  • Colin Johnson
    Leeds Institute of Molecular Medicine, University of Leeds, UK, United Kingdom
  • Robin Ali
    King’s College, London, UK, United Kingdom
  • Lyle Armstrong
    Biosciences Institute, Newcastle upon Tyne, Tyne and Wear, United Kingdom
  • Viktor Korolchuk
    Biosciences Institute, Newcastle upon Tyne, Tyne and Wear, United Kingdom
  • Henning Urlaub
    Max-Planck Institute of Biophysical Chemistry, Göttingen, Germany, Göttingen,, Germany
  • Sina Mozaffari-Jovin
    Max-Planck Institute of Biophysical Chemistry, Göttingen, Germany, Göttingen,, Germany
  • Majlinda Lako
    Biosciences Institute, Newcastle upon Tyne, Tyne and Wear, United Kingdom
  • Footnotes
    Commercial Relationships   Maria Georgiou None; Chunbo Yang None; Robert Atkinson None; Kuan-Ting Pan None; Adriana Buskin None; Marina Moya Molina None; Joseph Collin None; Sebastian E. J. Ludwig None; Sushma Nagaraja-Grellscheid None; Colin Johnson None; Robin Ali None; Lyle Armstrong None; Viktor Korolchuk None; Henning Urlaub None; Sina Mozaffari-Jovin None; Majlinda Lako None
  • Footnotes
    Support  Retina UK (GR595, GR584), MRC UK (MR/T017503/1), Fight for Sight (1456/1457) and ERC (CoG_614620).
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 3818. doi:
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      Maria Georgiou, Chunbo Yang, Robert Atkinson, Kuan-Ting Pan, Adriana Buskin, Marina Moya Molina, Joseph Collin, Sebastian E. J. Ludwig, Sushma Nagaraja-Grellscheid, Colin Johnson, Robin Ali, Lyle Armstrong, Viktor Korolchuk, Henning Urlaub, Sina Mozaffari-Jovin, Majlinda Lako; Activation of autophagy reverses progressive and deleterious protein aggregation in PRPF31 patient induced pluripotent stem cells (iPSCs)-derived retinal pigment epithelium (RPE) cells.. Invest. Ophthalmol. Vis. Sci. 2022;63(7):3818.

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

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Abstract

Purpose : Mutations in pre-mRNA processing factor 31 (PRPF31) result in autosomal-dominant retinitis pigmentosa (adRP) known as RP11, characterised by global dysregulation of spliceosome in retinal cells and the adjacent RPE cells. This study aims to investigate the disease pathomechanisms in Retinitis Pigmentosa caused by mutations in the PRPF31 gene.

Methods : Human iPSCs from three patients with severe and very severe PRPF31-adRP, unaffected individuals and a CRISPR/Cas9-corrected isogenic control were used to generate RPE monolayers. To fully assess the impacts of PRPF31 mutations, quantitative proteomics analyses of control and RP11-RPE cells and biochemical assays were performed.

Results : Quantitative proteomic analysis of control and patient RPE cells showed RNA splicing, retinoid metabolism and visual perception, and protein folding (UPR) pathways to be affected in RP11-RPE cells. The patient-derived RPE cells were characterised by reduced amounts of tri-snRNPs, spliceosome activity, and the presence of insoluble aggregates containing the mutant PRPF31 and misfolded, ubiquitin conjugated proteins, which accumulated progressively with time. The waste disposal mechanisms via autophagy and proteasome-mediated degradation were impaired, further exacerbating aggregate formation, which was closely linked with activation of cell death. Targeting the waste disposal mechanisms by activating the autophagy pathway using rapamycin resulted in the reduction of these cytoplasmic aggregates in RP11-RPE cells and improved cell survival.

Conclusions : Together these data indicate a vicious circle initiated by mutations in PRPF31, which lead to spliceosome dysregulation and accumulation of misfolded proteins in the form of insoluble cytoplasmic aggregates that affect RPE cell viability. Relieving the RPE cells from accumulation of these insoluble cytoplasmic aggregates presents a novel therapeutic strategy for RP11-patients.

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

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