Retinal Ganglion Cells With a Glaucoma OPTN(E50K) Mutation Exhibit Neurodegenerative Phenotypes when Derived from Three-Dimensional Retinal Organoids

• Published in: Stem cell reports Vol. 15; no. 1; pp. 52 - 66
• Main Authors: VanderWall, Kirstin B., Huang, Kang-Chieh, Pan, Yanling, Lavekar, Sailee S., Fligor, Clarisse M., Allsop, Anna R., Lentsch, Kelly A., Dang, Pengtao, Zhang, Chi, Tseng, Henry C., Cummins, Theodore R., Meyer, Jason S.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 14.07.2020; Elsevier
• Subjects: autophagy; CRISPR; differentiation; disease; gene editing; glaucoma; iPSC; modeling; neurodegeneration; retina; stem cell; CRISPR; autophagy; differentiation; disease; modeling; stem cell; retina; glaucoma; iPSC; neurodegeneration; gene editing
• ISSN: 2213-6711; 2213-6711
• DOI: 10.1016/j.stemcr.2020.05.009

Retinal ganglion cells (RGCs) serve as the connection between the eye and the brain, with this connection disrupted in glaucoma. Numerous cellular mechanisms have been associated with glaucomatous neurodegeneration, and useful cellular models of glaucoma allow for the precise analysis of degenerative phenotypes. Human pluripotent stem cells (hPSCs) serve as powerful tools for studying human disease, particularly cellular mechanisms underlying neurodegeneration. Thus, efforts focused upon hPSCs with an E50K mutation in the Optineurin (OPTN) gene, a leading cause of inherited forms of glaucoma. CRISPR/Cas9 gene editing introduced the OPTN(E50K) mutation into existing lines of hPSCs, as well as generating isogenic controls from patient-derived lines. RGCs differentiated from OPTN(E50K) hPSCs exhibited numerous neurodegenerative deficits, including neurite retraction, autophagy dysfunction, apoptosis, and increased excitability. These results demonstrate the utility of OPTN(E50K) RGCs as an in vitro model of neurodegeneration, with the opportunity to develop novel therapeutic approaches for glaucoma. [Display omitted] •CRISPR/Cas9 engineering of OPTN(E50K) glaucomatous hPSCs and isogenic controls•RNA sequencing displays important pathways linked to glaucomatous neurodegeneration•OPTN(E50K) RGCs encompass numerous neurodegenerative mechanisms In this article, VanderWall, Huang and colleagues present the extensive characterization of the OPTN(E50K) mutation in hPSC-derived RGCs using CRISPR/Cas9-engineered disease lines and isogenic controls. Results presented a detailed analysis of numerous neurodegenerative phenotypes observed in OPTN(E50K) RGCs and provide the opportunity to explore new targets for therapeutic intervention.