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Publication Detail
Regulation of retinal pigment epithelial cell phenotype by Annexin A8.
  • Publication Type:
    Journal article
  • Publication Sub Type:
    Article
  • Authors:
    Lueck K, Carr A-JF, Stampoulis D, Gerke V, Rescher U, Greenwood J, Moss SE
  • Publication date:
    05/07/2017
  • Pagination:
    4638
  • Journal:
    Scientific reports
  • Volume:
    7
  • Issue:
    1
  • Medium:
    Electronic
  • Print ISSN:
    2045-2322
  • Language:
    eng
  • Addresses:
    Department of Cell Biology, UCL Institute of Ophthalmology, 11-43 Bath Street, EC1V 9EL, London, United Kingdom. k.lueck@ucl.ac.uk.
Abstract
The retinoic acid derivative fenretinide (FR) is capable of transdifferentiating cultured retinal pigment epithelial (RPE) cells towards a neuronal-like phenotype, but the underlying mechanisms are not understood. To identify genes involved in this process we performed a microarray analysis of RPE cells pre- and post-FR treatment, and observed a marked down-regulation of AnnexinA8 (AnxA8) in transdifferentiated cells. To determine whether AnxA8 plays a role in maintaining RPE cell phenotype we directly manipulated AnxA8 expression in cultured and primary RPE cells using siRNA-mediated gene suppression, and over-expression of AnxA8-GFP in conjunction with exposure to FR. Treatment of RPE cells with AnxA8 siRNA recapitulated exposure to FR, with cell cycle arrest, neuronal transdifferentiation, and concomitant up-regulation of the neuronal markers calretinin and calbindin, as assessed by real-time PCR and immunofluorescence. In contrast, AnxA8 transient over-expression in ARPE-19 cells prevented FR-induced differentiation. Ectopic expression of AnxA8 in AnxA8-depleted cells led to decreased neuronal marker staining, and normal cell growth as judged by phosphohistone H3 staining, cell counting and cleaved caspase-3 levels. These data show that down-regulation of AnxA8 is both necessary and sufficient for neuronal transdifferentiation of RPE cells and reveal an essential role for AnxA8 as a key regulator of RPE phenotype.
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