July 2019
Volume 60, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2019
Differential transcriptome profile associated with pigmentation defects in induced pluripotent stem cell-derived retinal pigment epithelium from Albinism patients
Author Affiliations & Notes
  • Aman George
    Ophthalmic Genetics and Visual Function Branch, National Eye Institute/NIH, Bethesda, Maryland, United States
  • Fnu Ruchi
    Ophthalmic Genetics and Visual Function Branch, National Eye Institute/NIH, Bethesda, Maryland, United States
  • Tyler Pfister
    Ophthalmic Genetics and Visual Function Branch, National Eye Institute/NIH, Bethesda, Maryland, United States
  • David McGaughey
    Ophthalmic Genetics and Visual Function Branch, National Eye Institute/NIH, Bethesda, Maryland, United States
  • Nathan Hotaling
    Ophthalmic Genetics and Visual Function Branch, National Eye Institute/NIH, Bethesda, Maryland, United States
  • Mones S Abu-Asab
    Ophthalmic Genetics and Visual Function Branch, National Eye Institute/NIH, Bethesda, Maryland, United States
  • Qin Wan
    Ophthalmic Genetics and Visual Function Branch, National Eye Institute/NIH, Bethesda, Maryland, United States
  • Congxiao Zhang
    Ophthalmic Genetics and Visual Function Branch, National Eye Institute/NIH, Bethesda, Maryland, United States
  • Kapil Bharti
    Ophthalmic Genetics and Visual Function Branch, National Eye Institute/NIH, Bethesda, Maryland, United States
  • Brian Patrick Brooks
    Ophthalmic Genetics and Visual Function Branch, National Eye Institute/NIH, Bethesda, Maryland, United States
  • Footnotes
    Commercial Relationships   Aman George, None; Fnu Ruchi, None; Tyler Pfister, None; David McGaughey, None; Nathan Hotaling, None; Mones Abu-Asab, None; Qin Wan, None; Congxiao Zhang, None; Kapil Bharti, None; Brian Brooks, None
  • Footnotes
    Support  Knights Templar Eye Foundation, NIH Intramural
Investigative Ophthalmology & Visual Science July 2019, Vol.60, 2877. doi:https://doi.org/
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    • Get Citation

      Aman George, Fnu Ruchi, Tyler Pfister, David McGaughey, Nathan Hotaling, Mones S Abu-Asab, Qin Wan, Congxiao Zhang, Kapil Bharti, Brian Patrick Brooks; Differential transcriptome profile associated with pigmentation defects in induced pluripotent stem cell-derived retinal pigment epithelium from Albinism patients. Invest. Ophthalmol. Vis. Sci. 2019;60(9):2877. doi: https://doi.org/.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Oculocutaneous albinism (OCA) is a genetically heterogenous group of conditions that affect pigmentation and vision. Previous studies have suggested a correlation between the extent of pigmentation loss and vision defects. We hypothesized that if the pathophysiology of vision defects in two forms of albinism (OCA1A and OCA2) is shared, then we should observe events at the molecular level that are present in the albinism group but not in control. To test our hypothesis, we developed a “disease in a dish” model of OCA using patient-derived retinal pigment epithelium (RPE) cells and identify transcriptional level changes that occur in RPE of OCA1A and OCA2 patients but not in unaffected individuals.

Methods :
Induced pluripotent stem cells (iPSCs) derived from albinism patients (OCA1A and OCA2) and unaffected individuals (control) were differentiated into RPE and characterized by trans-epithelial resistance (TER) measurements, immunostaining, photoreceptor outer segment (OS) phagocytosis assay and electron microscopy (EM). Eight-week post seeding of RPE cells on trans-wells, RNA was isolated and Poly(A)-enriched mRNA sequencing was performed on the Illumina platform. Gene- and transcript-level expression was quantified with quasi-mapping (Salmon) and differential expression was calculated with DESeq2.

Results : RPE derived from albinism patient iPSCs (OCA-RPE) and control iPSCs (control-RPE) were similar in terms of cellular morphology, TER, polarization and ability to phagocytose photoreceptor OS. EM revealed pigmentation defects in OCA-RPE that were not observed in control-RPE cells. Analysis of the transcriptome profiles identified significant differences (p≤ 0.05) between albinism and control-RPE. OCA-RPE and control-RPE samples clustered independently on the Principal Component Analysis (PCA) plot. Gene Ontology (GO) and Gene Set enrichment analyses revealed the “Active transmembrane transporters” gene family and “Retinol metabolism” pathways being significantly different from control in both albinism RPE lines.

Conclusions : We identified shared pathways at the transcriptional level that are significantly affected in two forms of congenital albinism caused by mutations in two different genes. Studies are underway to investigate if these pathways contribute to vision-related conditions observed in albinism patients.

This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.

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