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  Activation of autophagy reverses progressive and deleterious protein aggregation in PRPF31 patient‐induced pluripotent stem cell‐derived retinal pigment epithelium cells

Georgiou, M., Yang, C., Atkinson, R., Pan, K., Buskin, A., Molina, M. M., et al. (2022). Activation of autophagy reverses progressive and deleterious protein aggregation in PRPF31 patient‐induced pluripotent stem cell‐derived retinal pigment epithelium cells. Clinical and Translational Medicine, 12(3): e759. doi:10.1002/ctm2.759.

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 Creators:
Georgiou, M., Author
Yang, C., Author
Atkinson, R., Author
Pan, K.‐T., Author
Buskin, A., Author
Molina, M. M., Author
Collin, J., Author
Al‐Aama, J., Author
Goertler, F., Author
Ludwig, S. E. J., Author
Davey, T., Author
Lührmann, R.1, Author           
Nagaraja‐Grellscheid, S., Author
Johnson, C. A., Author
Ali, R., Author
Armstrong, L., Author
Korolchuk, V., Author
Urlaub, H.2, Author           
Mozaffari‐Jovin, S., Author
Lako, M., Author
Affiliations:
1Emeritus Group of Cellular Biochemistry, Max Planck Institute for Multidisciplinary Sciences, Max Planck Society, ou_3350136              
2Research Group of Bioanalytical Mass Spectrometry, Max Planck Institute for Multidisciplinary Sciences, Max Planck Society, ou_3350290              

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Free keywords: aggregate formation, autophagy, human pluripotent stem cells, proteasome, PRPF31, retinal organoids, retinitis pigmentosa, RPE, tri-snRNP assembly defects, UPR
 Abstract: Abstract Introduction: Mutations in pre-mRNA processing factor 31 (PRPF31), a core protein of the spliceosomal tri-snRNP complex, cause autosomal-dominant retinitis pigmentosa (adRP). It has remained an enigma why mutations in ubiqui- tously expressed tri-snRNP proteins result in retina-specific disorders, and so far, the underlying mechanism of splicing factors-related RP is poorly understood. Methods: We used the induced pluripotent stem cell (iPSC) technology to gen- erate retinal organoids and RPE models from four patients with severe and very severe PRPF31-adRP, unaffected individuals and a CRISPR/Cas9 isogenic con- trol. Results: To fully assess the impacts of PRPF31 mutations, quantitative pro- teomics analyses of retinal organoids and RPE cells were carried out showing RNA splicing, autophagy and lysosome, unfolded protein response (UPR) and visual cycle-related pathways to be significantly affected. Strikingly, the patient- derived RPE and retinal cells were characterised by the presence of large amounts of cytoplasmic aggregates containing the mutant PRPF31 and mis- folded, ubiquitin-conjugated proteins including key visual cycle and other RP- linked tri-snRNP proteins, which accumulated progressively with time. The mutant PRPF31 variant was not incorporated into splicing complexes, but reduc- tion of PRPF31 wild-type levels led to tri-snRNP assembly defects in Cajal bod- ies of PRPF31 patient retinal cells, altered morphology of nuclear speckles and reduced formation of active spliceosomes giving rise to global splicing dysregu- lation. Moreover, the impaired waste disposal mechanisms further exacerbated aggregate formation, and targeting these by activating the autophagy pathway using Rapamycin reduced cytoplasmic aggregates, leading to improved cell sur- vival. Conclusions: Our data demonstrate that it is the progressive aggregate accumu- lation that overburdens the waste disposal machinery rather than direct PRPF31- initiated mis-splicing, and thus relieving the RPE cells from insoluble cytoplas- mic aggregates presents a novel therapeutic strategy that can be combined with gene therapy studies to fully restore RPE and retinal cell function in PRPF31- adRP patients.

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Language(s): eng - English
 Dates: 2022-03-162022-03
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1002/ctm2.759
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Title: Clinical and Translational Medicine
Source Genre: Journal
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Pages: 27 Volume / Issue: 12 (3) Sequence Number: e759 Start / End Page: - Identifier: ISSN: 2001-1326
ISSN: 2001-1326