July 2019
Volume 60, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2019
Structural and functional alterations at the photoreceptor-RPE interface lead to disease-associated pathological changes in a hiPSC-derived model of an inherited retinal degenerative disease.
Author Affiliations & Notes
  • Ruchira Singh
    Opthalmology and Biomedical Genetics, University of Rochester, Rochester, New York, United States
  • Cynthia Tang
    Ophthalmology, University of Rochester Medical Center, Rochester, New York, United States
  • Sonal Dalvi
    Ophthalmology, University of Rochester Medical Center, Rochester, New York, United States
  • Lauren Winschel
    Ophthalmology, University of Rochester Medical Center, Rochester, New York, United States
  • Jimin Han
    Ophthalmology, University of Rochester Medical Center, Rochester, New York, United States
  • Chad A Galloway
    Ophthalmology, University of Rochester Medical Center, Rochester, New York, United States
  • Celia Soto
    Ophthalmology, University of Rochester Medical Center, Rochester, New York, United States
  • Leslie A MacDonald
    Ophthalmology, University of Rochester Medical Center, Rochester, New York, United States
  • Footnotes
    Commercial Relationships   Ruchira Singh, None; Cynthia Tang, None; Sonal Dalvi, None; Lauren Winschel, None; Jimin Han, None; Chad Galloway, None; Celia Soto, None; Leslie MacDonald, None
  • Footnotes
    Support  BrightFocus Foundation Macular Degeneration Grant, Foundation of Fighting Blindness Individual Investigator Award, National Institute of Health, NIH-1R01EY028167, and Research to Prevent Blindness, RPB’s Career Development Award, RPB's Unrestricted Challenge Grant to Department of Ophthalmology at University of Rochester
Investigative Ophthalmology & Visual Science July 2019, Vol.60, 4763. doi:
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    • Get Citation

      Ruchira Singh, Cynthia Tang, Sonal Dalvi, Lauren Winschel, Jimin Han, Chad A Galloway, Celia Soto, Leslie A MacDonald; Structural and functional alterations at the photoreceptor-RPE interface lead to disease-associated pathological changes in a hiPSC-derived model of an inherited retinal degenerative disease.. Invest. Ophthalmol. Vis. Sci. 2019;60(9):4763.

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

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Abstract

Purpose : Juvenile neuronal ceroid lipofuscinosis (JNCL, Batten disease) is caused by mutation in CLN3 gene. Furthermore, vision loss caused by retinal degeneration is a characteristic clinical feature of JNCL. Although retinal degeneration in JNCL is linked to photoreceptor cell loss, both retinal pigment epithelium (RPE) and photoreceptors are severely affected in JNCL. The purpose of this study was to utilize human induced pluripotent stem cell (hiPSC)-based disease modeling to determine the role of primary RPE dysfunction and photoreceptor-RPE interaction in the retinal pathophysiology of JNCL.

Methods : hiPSCs derived from JNCL patients and unaffected controls were differentiated to hiPSC-RPE and neural retina (NR). Next, morphological, gene and protein expression and functional characteristics of mature monocultures of control vs. JNCL hiPSC-RPE were evaluated. Furthermore, photoreceptor outer segment (POS) phagocytosis and degradation after acute POS feeding (2h) and the pattern of autofluorescence accumulation (cell surface vs. intracellular) after chronic POS feeding (daily for 2wks) was compared in control vs. JNCL hiPSC-RPE cultures

Results : In the absence of exogenous stressors, JNCL hiPSC-RPE and NR cultures did not display any disease-associated cellular phenotypes. In contrast, when exposed to a physiological dose of POS (~20POS/RPE/day), reduced uptake of POS was seen in JNCL-hiPSC-RPE cells compared to control hiPSC-RPE cells. Furthermore, consistent with reduced accumulation of autofluorescent material (by-products of POS digestion) in the RPE layer and increased autofluorescence in the photoreceptor layer of JNCL patient eyes, decreased intracellular autofluorescence and conversely increased cell surface autofluorescence was seen in JNCL hiPSC-RPE cells compared to control hiPSC-RPE cells after chronic POS feeding (daily for 2wks). Additionally, consistent with a POS phagocytosis defect, JNCL hiPSC-RPE displayed microvilli disorganization/ballooning.

Conclusions : Using hiPSC-based disease modeling, we show that CLN3 mutation in JNCL affect key RPE structure (microvilli) and function (phagocytosis of POS) and instigate autofluorescence changes in JNCL retina/RPE. Furthermore, our data implicate primary RPE dysfunction and impaired photoreceptor-RPE interface in JNCL retinal pathology.

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

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