Pluto Bioinformatics

GSE145069: Differential gene expression and pathway analysis by RNA sequencing in human OPTN(E50K) retinal ganglion cells

Bulk RNA sequencing

Retinal ganglion cells (RGCs) serve as the essential connection between the eye and the brain, with this connection disrupted in blinding disorders such as glaucoma. Numerous cellular mechanisms have been associated with glaucomatous neurodegeneration, and useful models for the study of glaucoma allow for the precise analysis of these degenerative phenotypes. Human pluripotent stem cells (hPSCs) serve as powerful tools for the in vitro analysis of human neurodegenerative diseases, particularly those cellular mechanisms underlying degeneration. Thus, efforts of the current study were initially focused upon the use of hPSCs with an E50K mutation in the Optineurin (OPTN) gene underlying glaucomatous neurodegeneration. CRISPR/Cas9 gene editing approaches were used to introduce the OPTN(E50K) mutation into existing lines of hPSCs, as well as the generation of isogenic control lines from existing OPTN(E50K) patient-derived hPSC lines. When grown from glaucomatous sources, hPSC-derived RGCs developed similarly to isogenic controls. Conversely, at later stages of maturation, OPTN(E50K) RGCs exhibited numerous deficits associated with a neurodegenerative phenotype, including neurite retraction, autophagy pathway disruption, apoptosis, and increased excitability. The results of this study provide an extensive analysis of the OPTN(E50K) mutation in hPSC-derived RGCs, with a number of biochemical and molecular alterations associated with functional changes to these RGCs. The opportunity now exists to further explore the precise cellular pathways leading to RGC death in glaucoma, as well as develop novel translational approaches for glaucoma including pharmacological screening and cell replacement therapies. SOURCE: Xiaoyu Lu INDIANA UNIVERSITY