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Generation and characterization of retinal pigment epithelium from patient iPSC line to model oculocutaneous albinism (OCA)1A disease

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Abstract

Oculocutaneous albinism (OCA) is characterized by reduced melanin biosynthesis affecting the retina, thus impairing visual function. The disease pathology of OCA is poorly understood at the cellular level due to unavailability of suitable biological model systems. This study aimed to develop a disease-specific in vitro model for OCA type 1A, the most severe form caused by TYR (tyrosinase) gene mutations, using retinal pigment epithelium (RPE) differentiated from patient-derived human-induced pluripotent stem cells (hiPSCs). A comparative study between healthy and OCA1A RPE cells revealed that while healthy RPE cells exhibited timely onest of pigmentation during differentiation, OCA1A RPE cells failed to pigment even after an extended culture period. This observation was validated by ultrastructural studies using electron microscopy, hinting at melanosome-specific defects. Immunocytochemistry demonstrated abnormal expression patterns of melanogenesis-specific protein markers in OCA1A RPE cells, indicating reduced or absence of melanin synthesis. Next, a quantitative assay was performed to confirm the absence of melanin production in OCA1A RPE cells. Tyrosinase assay showed no activity in OCA1A compared with healthy RPE, suggesting non-functionality of TYR, further corroborated by western blot analysis showing complete absence of the protein. Gene expression by RNA sequencing of healthy and OCA1A RPE cells uncovered differential gene expression associated with lens development, visual perception, transmembrane transporter activity, and key signaling pathways. This disease-in-a-dish model of OCA1A provides an excellent platform to understand disease mechanism, identify potential therapeutic targets, and facilitate gene therapy or gene correction.

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Acknowledgements

The authors thank the Next Generation Genomics Facility (NGGF) and the Central Imaging and Flow Cytometry Facility (CIFF) at the Bangalore LifeScience Cluster. Jasmin MR from the Centre for Cellular and Molecular Platforms (C-CAMP), Bengaluru, is acknowledged for her technical help with Western blots. The authors are grateful to Jair Montford, National Eye Institute, NIH, Bethesda, for the electron microscopy work. The authors are also thankful to their collaborators, Prof. Ramaswamy Subramanian and Dr. Kanaga Vijayan, Purdue University, Indiana, for their inputs during the study.

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Authors and Affiliations

  1. Centre for Cellular and Molecular Platforms (C-CAMP), GKVK Post, Bellary Road, Bengaluru, 560065, India

    Janavi Subramani & Taslimarif Saiyed

  2. Eyestem Research, GKVK Post, Bellary Road, Bengaluru, 560065, India

    Niharika Patlolla & Rajarshi Pal

  3. Centre for Eye Genetics and Research, Yashoda Nagar, Bengaluru, 560064, India

    Rajani Battu

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  1. Janavi Subramani
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  2. Niharika Patlolla
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  3. Rajani Battu
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  4. Taslimarif Saiyed
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  5. Rajarshi Pal
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Correspondence to Rajarshi Pal.

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Corresponding editor: Sudha Bhattacharya

This article is part of the Topical Collection: The Rare Genetic Disease Research Landscape in India.

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Subramani, J., Patlolla, N., Battu, R. et al. Generation and characterization of retinal pigment epithelium from patient iPSC line to model oculocutaneous albinism (OCA)1A disease. J Biosci 49, 21 (2024). https://doi.org/10.1007/s12038-023-00406-7

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  • DOI: https://doi.org/10.1007/s12038-023-00406-7

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