September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
NAMPT inhibitors are Selectively Toxic to Human Pluripotent Stem Cells (hPSC) and can Minimize Tumorigenic Potential of hPSC-derived RPE cells
Author Affiliations & Notes
  • Qinghui Hu
    School of Biomedical Sciences, The University of Hong Kong, Hong Kong, Hong Kong
    LKS Faculty of Medicine, The University of Hong Kong, Hong Kong, Hong Kong
  • Amy CY Lo
    Department of Ophthalmology, The University of Hong Kong, Hong Kong, Hong Kong
    LKS Faculty of Medicine, The University of Hong Kong, Hong Kong, Hong Kong
  • David Wong
    LKS Faculty of Medicine, The University of Hong Kong, Hong Kong, Hong Kong
  • Kenneth Richard Boheler
    School of Biomedical Sciences, The University of Hong Kong, Hong Kong, Hong Kong
    LKS Faculty of Medicine, The University of Hong Kong, Hong Kong, Hong Kong
  • Footnotes
    Commercial Relationships   Qinghui Hu, None; Amy Lo, None; David Wong, None; Kenneth Boheler, None
  • Footnotes
    Support  Seed Funding Programme for Basic Research at HKU and Health and Medical Research Fund, Hong Kong
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 6544. doi:
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    • Get Citation

      Qinghui Hu, Amy CY Lo, David Wong, Kenneth Richard Boheler; NAMPT inhibitors are Selectively Toxic to Human Pluripotent Stem Cells (hPSC) and can Minimize Tumorigenic Potential of hPSC-derived RPE cells. Invest. Ophthalmol. Vis. Sci. 2016;57(12):6544.

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

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Abstract

Purpose : This study aims to determine the ability of nicotinamide phosphoribosyltransferase (NAMPT) inhibitors to selectively eliminate undifferentiated and tumorigenic cells from hPSC cultures differentiating to retinal pigmented epithelial (RPE) cells.

Methods : We cultured and differentiated hPSCs to neural progenitors and RPE cells, and studied ARPE19 cells. STF31 and FK866, inhibitors of the NAD salvage pathway protein NAMPT, were added in a dose- and time-dependent manner and cell viability assays (Neutral Red and XTT) performed . Teratoma assays are performed in NOD-SCID mice.

Results : Human PSCs differentiated in vitro to RPE cells exhibit epithelial cell morphology, visible pigment from week 4 of differentiation, and contained RNA (BEST1, MITF) markers typical of RPE cells. The putative hPSC-derived RPE cell populations are ~70% pure, and work is on-going to improve RPE purity for functional assessments and for viability testing. To evaluate selective toxicity of NAMPT inhibitors on cells of the neuroectoderm lineage, human ES cells (H7), iPS cells (KB3), PAX 6-positive neural progenitors and ARPE 19 cells were treated. NAMPT inhibitors were highly toxic to hPSCs (0-5% viable cells) within 48-72 hours. PAX6 positive cells showed some decrease in viability (50-60% viable cells) at 72 hours. ARPE19 cell toxicity was minimal. Teratoma assays following 24-hour treatment of hPSCs did not completely prevent hPSC tumour formation, but longer treatment times appear effective.

Conclusions : Elimination of tumorigenicity is a prerequisite for clinical intervention. NAMPT shows selective cell toxicity: hPSCs>>neural progenitors> ARPE19 cells. Purified hPSC-derived RPEs are likely to show limited toxicity to NAMPT, suggesting that treatment of these cultures with NAMPT inhibitors prior to transplantation will effectively eliminate all tumorigenic potential from these cultures – thus advancing efforts to treat Aging-related Macular Degeneration in humans.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

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