GSE210331
Transcriptional comparison of adult human primary Retinal Pigment Epithelium, human pluripotent stem cell-derived Retinal Pigment Epithelium, and ARPE19 cells
The therapeutic potential of pluripotent stem cells is great as they promise to Q7  usher in a new era of medicine where cells or organs may be prescribed to  replace dysfunctional tissue. At the forefront are efforts in the eye to develop Q8  this technology as it lends itself to in vivo monitoring and sophisticated imaging  modalities. In the retina, retinal pigment epithelium (RPE) is the most promising  replacement cell as it has a single layer, is relatively simple to transplant, and is  associated with dozens of eye diseases. However, after transplantation, they  may transform and cause complications. This transformation may be partially  due to incomplete maturation. With the goal of learning how to mature RPE, we  compared iPSC-RPE with adult primary RPE and the immortalized ARPE-19 line.  We cultured ARPE-19, iPSC-RPE, and adult human RPE for one month,  evaluating morphology, RPE marker staining, and transepithelial electrical  resistance as quality control indicators. We then isolated RNA for bulk RNAsequencing  and DNA for genotyping. We genotyped adult RPE lines for the top  age-related macular degeneration (AMD) and proliferative vitreoretinopathy  (PVR) risk allele polymorphisms. Transcriptome data verified that both adult and  iPSC-RPE exhibit similar RPE gene expression signatures, significantly more  than ARPE-19. In addition, in iPSC-RPE, gene networks relating to epithelial to  mesenchymal transition (EMT), stem cell maintenance, retina development, and  muscle contraction were significantly elevated compared to those of adult RPE.  We compared adult human RPE to iPSC-RPE in a model of EMT and observed an  increased sensitivity of iPSC-RPE to producing contractile aggregates in vitro  which resembles incident reports upon transplantation. P38 inhibition was  capable of inhibiting iPSC-RPE–derived aggregates. In summary, we find  that the transcriptomic signature of iPSC-RPE conveys an immature RPE state  which may be ameliorated by targeting “immature” gene regulatory networks. Overall design: Comparative gene expression profiling by RNASeq of ahRPE, iPSC-RPE and ARPE19 cells.