GSE137910
Morphological and Molecular Defects in Human Three-Dimensional Retinal Organoid Model of X-Linked Juvenile Retinoschisis
The goals of this study are to analyze the effects of RS1 mutation on retinal transcriptome profiling during retina differentiation. We used next generation sequencing to analyze the retinal mRNA profiles in a panel of time-course differentiated 3D retinal cups (RCs) from control donor- and XLRS patient-derived iPSCs, as well as 150-day differentiated RCs from CRISPR/Cas9-reparied patient-derived iPSCs. The transcriptomes of control-RCs and patient-RCs considerably diverged along the time course of differentiation, with the difference becoming more pronounced at the later time points. In contrast, analysis of repaired RCs revealed transcriptomes that were highly similar to those of control-RCs and significantly different from those of patient-RCs. Because RS1 is only expressed in later stage of differentiation, we focued on days 90, 120, and 150 post-induction to identify the genes whose expression was reduced at least 2-fold in patient-RCs. Among all of these genes, 326 genes were overexpressed in normal RCs at all of these stages, emphasizing their importance. According to GSEA annotation, nine out of these 326 genes were implicated in eye development. Among these nine genes, the expression of RS1, IQCB1, and OPA1 was rescued in CRISPR repaired RCs. The relative loss of expression became even more prominent in later stages of retinal differentiation. Our transcriptome analysis revealed significant changes in gene expression between patient-derived and control-RCs that become more pronounced at the later stages of development. This implies that RS1 is essential for normal retinal development at the systemic level. These results reflect the progressive nature of the disease phenotypes observed in RS-1-mutant RCs, mirror the observed ciliary phenotype, and reveal potential links of XLRS to other hereditary retinopathies. Overall design: mRNA profiles of 3D retinal organoids at Day 0, 50, 90, 120, 150 post-differentiation induction from control donor- and XLRS patient-derived iPSCs, as well as from Day 150 3D retinal organoids differentiated from CRISPR/Cas9-corrected patient-derived iPSCs, were generated by deep sequencing using Illumina HiSeq 2000.