Human iPSC-Derived RPE and Retinal Organoids Reveal Impaired Alternative Splicing of Genes Involved in Pre-mRNA Splicing in PRPF31 Autosomal Dominant Retinitis Pigmentosa Type 11
Mutations in pre-mRNA processing factors (PRPFs) cause 40% of autosomal dominant retinitis pigmentosa (RP), but it is unclear why mutations in ubiquitously expressed PRPFs cause retinal disease. To understand the molecular basis of this phenotype, we have generated RP type 11 (PRPF31-mutated) patient-specific retinal organoids and retinal pigment epithelium (RPE) from induced pluripotent stem cells (iPSC). Impaired alternative splicing of genes encoding pre-mRNA splicing proteins occurred in patient-specific retinal cells and Prpf31 /- mouse retinae, but not fibroblasts and iPSCs, providing mechanistic insights into retinal-specific phenotypes of PRPFs. RPE was the most affected, characterised by loss of apical-basal polarity, reduced trans-epithelial resistance, phagocytic capacity, microvilli, and cilia length and incidence. Disrupted cilia morphology was observed in patient-derived-photoreceptors that displayed progressive features associated with degeneration and cell stress. In situ gene-editing of a pathogenic mutation rescued key structural and functional phenotypes in RPE and photoreceptors, providing proof-of-concept for future therapeutic strategies.
Buskin, Adriana and Zhu, Lili and Chichagova, Valeria and Basu, Basudha and Mozaffari‐Jovin, Sina and Dolan, David and Droop, Alastair and Colli, Joseph and Bronstei, Revital and Mehrotr, Sudeep and Farkas, Michael and Hilgen, Gerrit and White, Kathryn and Hallam, Dean and Bialas, Katarzyna and Chung, Git and Mellough, Carla and Ding, Yuchun and Krasnogor, Natalio and Przyborski, Stefan and Al‐Aama, Jumana and Alharthi, Sameer and Xu, Yaobo and Wheway, Gabrielle and Szymanska, Katarzyna and McKibbin, Martin and Inglehearn, Chris F and Elliott, David J and Lindsay, Susan and Ali, Robin R and Steel, David H and Armstrong, Lyle and Sernagor, Evelyne and Pierce, Eric and Lührmann, Reinhard and Grellscheid, Sushma‐Nagaraja and Johnson, Colin A and Lako, Majlinda, Human iPSC-Derived RPE and Retinal Organoids Reveal Impaired Alternative Splicing of Genes Involved in Pre-mRNA Splicing in PRPF31 Autosomal Dominant Retinitis Pigmentosa Type 11 (2018). Available at SSRN: https://ssrn.com/abstract=3155753 or http://dx.doi.org/10.2139/ssrn.3155753
This version of the paper has not been formally peer reviewed.
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