Functional Assessment of Patient-Derived Retinal Pigment Epithelial Cells Edited by CRISPR/Cas9.

TitleFunctional Assessment of Patient-Derived Retinal Pigment Epithelial Cells Edited by CRISPR/Cas9.
Publication TypeJournal Article
Year of Publication2018
AuthorsFoltz LP, Howden SE, Thomson JA, Clegg DO
JournalInt J Mol Sci
Date Published2018 Dec 19

Retinitis pigmentosa is the most common form of inherited blindness and can be caused by a multitude of different genetic mutations that lead to similar phenotypes. Specifically, mutations in ubiquitously expressed splicing factor proteins are known to cause an autosomal dominant form of the disease, but the retina-specific pathology of these mutations is not well understood. Fibroblasts from a patient with splicing factor retinitis pigmentosa caused by a missense mutation in the splicing factor were used to produce three diseased and three CRISPR/Cas9-corrected induced pluripotent stem cell (iPSC) clones. We differentiated each of these clones into retinal pigment epithelial (RPE) cells via directed differentiation and analyzed the RPE cells in terms of gene and protein expression, apicobasal polarity, and phagocytic ability. We demonstrate that RPE cells can be produced from patient-derived and corrected cells and they exhibit morphology and functionality similar but not identical to wild-type RPE cells in vitro. Functionally, the RPE cells were able to establish apicobasal polarity and phagocytose photoreceptor outer segments at the same capacity as wild-type cells. These data suggest that patient-derived iPSCs, both diseased and corrected, are able to differentiate into RPE cells with a near normal phenotype and without differences in phagocytosis, a result that differs from previous mouse models. These RPE cells can now be studied to establish a disease-in-a-dish system relevant to retinitis pigmentosa.

Alternate JournalInt J Mol Sci
PubMed ID30572641
Grant ListDR1-01444, CL1-00521, TB1-01177, FA1-00616 and TG2-01151 / / California Institute for Regenerative Medicine /
N/A / / The Garland Initiative for Vision, The Breaux Foundation, and the Foundation Fighting Blindness Wynn-Gund Translational Research Acceleration Program /