Differential gene expression analysis of wild type and heterozygous RB1 mutant human iPSC derived retinal organoids.

Recent in vitro studies using RB1+/- fibroblasts and MSCs have shown molecular and functional disruptions without the need for biallelic loss of RB1. However, this was not reflected in the recent in vitro studies employing RB1+/- retinal organoids. To gain further insights into the molecular disruptions in the RB1+/- retinal organoids we performed a high throughput RNA-sequencing analysis.iPSCs were generated from RB1+/+ and RB1+/- Orbital adipose mesenchymal stem cells (OAMSCs) derived from retinoblastoma patients. RB1+/+ and RB1+/- iPSCs were subjected to step-wise retinal differentiation protocol and high throughput RNA-sequencing followed by differential gene expression analysis and Gene set enrichment analysis (GSEA) was performed.The analysis revealed that even though there are no gross observable differences, subtle molecular changes in RB1+/- retinal organoids were observed. We report that there is mild shift from the regular metabolic process of glycolysis to oxidative phosphorylation in the RB1+/- retinal organoids which could be setting a premise for tumorigenesis. RNA sequencing was performed on the RB-iPSC derived retinal organoids