June 2023
Volume 64, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2023
A retinal organoid model of retinoblastoma
Author Affiliations & Notes
  • Gail M Seigel
    Communicative Disorders and Sciences, University at Buffalo, Buffalo, New York, United States
  • M Natalia Vergara
    Ophthalmology, University of Colorado Anschutz Medical Campus, Aurora, Colorado, United States
  • Kiarra Furey
    Communicative Disorders and Sciences, University at Buffalo, Buffalo, New York, United States
  • Dhavalkumar Shah
    Pharmaceutical Sciences, University at Buffalo, Buffalo, New York, United States
  • Footnotes
    Commercial Relationships   Gail Seigel None; M Natalia Vergara None; Kiarra Furey None; Dhavalkumar Shah None
  • Footnotes
    Support  This research was funded in part by The Childhood Eye Cancer Trust (Dr. Judith Kingston Research Fund), The Developmental Studies Hybridoma Bank at the University of Iowa
Investigative Ophthalmology & Visual Science June 2023, Vol.64, 1317. doi:
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    • Get Citation

      Gail M Seigel, M Natalia Vergara, Kiarra Furey, Dhavalkumar Shah; A retinal organoid model of retinoblastoma. Invest. Ophthalmol. Vis. Sci. 2023;64(8):1317.

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

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Abstract

Purpose : Induced pluripotent stem cells (iPSCs) can recapitulate early stages of retinal development, a time when retinoblastoma (RB) tumors can initiate in vivo. In this study, we tested the hypothesis that hybrid RB-iPSC-derived retinal spheroids would represent an ex-vivo 3-D model of retinoblastoma that could be used as a novel system to study RB tumor progression, with the potential for use in testing RB-targeted therapies.

Methods : We performed co-culture studies combining iPSC-derived retinal organoids and human RB cells with low (WERI-RB27 cells) and high (Y79 cells) invasive/metastatic potential. Human iPSCs were differentiated into retinal structures for 120 days prior to three weeks of co-culture with RB cells. Co-cultures consisted of five control retinal organoid cultures alone, four co-cultures of retinal organoids + WERI- RB27 cells, and four co-cultures of retinal organoids + Y79 cells. After three weeks of co-culture, hybrid RB-retinal and control organoids were fixed and prepared as frozen sections for immunohistochemical analysis (structural markers MAP2 and Nestin and the glial marker Glutamine Synthetase), visualized by fluorescence microscopy, with fluorescence intensity analyzed by FIJI image analysis.

Results : Control retinal organoid cultures (without RB cells) exhibited normal histoarchitecture and immunoreactivity for the markers of interest. Both RB cell lines disrupted the organization of these retinal organoids in a manner consistent with metastasis and invasion. This disruption was accompanied by statistically significant decreases in immunoreactivity of Glutamine Synthetase (Control vs. WERI-RB27 and Y79 p<0.001), MAP2 (Control vs. WERI-RB27 and Y79 p<0.01) and Nestin (Control vs. Y79 p< 0.001).

Conclusions : We have shown that iPSC-derived retinal organoids can be successfully co-cultured with RB cells, resulting in disorganized retinal histoarchitecture and altered immunoreactivity to structural proteins such as MAP2 and Nestin, as well as the retinal glial marker glutamine synthetase. Results from our study may lead to a useful ex-vivo model system that will provide a platform for assessing retinoblastoma tumor progression and personalized medicine approaches using patient-derived tumor cells.

This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.

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