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Proteomic Profiling of iPSC-Derived Retinal Pigment Epithelium following Ferric Ammonium Citrate Treatment

ABSTRACT: iPSC-derived retinal pigment epithelium (iPSC-RPE) cells were cultured in vitro under standard conditions. The cells were then subjected to either a control treatment or treatment with 200 µM ferric ammonium citrate (FAC) for a period of 10 days to induce iron overload. Following the treatment period, total cellular proteins were harvested from each group. Protein samples were subsequently prepared and analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) for global proteomic profiling. The experiment included two independent biological replicates for both the control and FAC-treated conditions to ensure statistical robustness.

ORGANISM(S): Homo Sapiens

SUBMITTER: Haibin Tian

PROVIDER: PXD072937 | iProX | Sun Jan 11 00:00:00 GMT 2026

REPOSITORIES: iProX

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Project description: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.

Dataset Information

Proteomic Profiling of iPSC-Derived Retinal Pigment Epithelium following Ferric Ammonium Citrate Treatment

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