Differentiation of human retinal pigment epithelial cells into neuronal phenotype by N‐(4‐hydroxyphenyl)retinamide

Abstract

ARPE-19, a human retinal pigment epithelial (RPE) cell line, has been widely used in studies of RPE function as well as gene expression. Here, we report the novel finding that N-(4-hydroxyphenyl)retinamide (fenretinide), a synthetic retinoic acid derivative and a potential chemopreventive agent against cancer, induced the differentiation of ARPE-19 cells into a neuronal phenotype. The treated cells lost their epithelial phenotype and exhibited a typical neuronal shape with long processes (four to five times longer than the cell body). The onset of fenretinide-induced neuronal differentiation was dose and time dependent, started within 1-2 days, and lasted at least 4 weeks. Immunohistochemical studies indicated that the expression of neurofilament proteins (NF160 and NF200), calretinin and neural cell adhesion molecule was increased in these differentiated cells. Western blot analysis indicated that cellular retinaldehyde-binding protein, which is normally expressed in RPE cells, was decreased in treated cells. Protein analysis on a two-dimensional gel followed by matrixassisted laser desorption ionization-time of flight mass spectrometric analysis demonstrated that heat-shock protein 70 was increased after fenretinide treatment. Thus, fenretinide, a synthetic retinoid, is able to induce neuronal differentiation of human RPE cells in culture.

Identification of a Lipase-linked Cell Membrane Receptor for Pigment Epithelium-derived Factor

“…obtained with the antibody to the extracellular peptide region R A in nonpermeabilized cells. The staining in ARPE-19 cells was visibly more intense around the nuclei where the density/ thickness of the cell is increased, and similar to that reported for N-CAM, a surface marker for ARPE-19 (55). Cells without Fl-PEDF did not show fluorescent signal.…”

The Effect of Am-80, a Synthetic Retinoid, on Spinal Cord Injury-Induced Motor Dysfunction in Rats

“…Promotion of neuronal differentiation by another retinoid derivative has already been reported, 41) as well as by other selective agonists. 24,38) Cell replacement therapy is currently attracting attention as a potentially attractive strategy for the treatment of neurodegenerative diseases including SCI.…”

N‐(4‐hydroxyphenyl)retinamide induces apoptosis in human retinal pigment epithelial cells: Retinoic acid receptors regulate apoptosis, reactive oxygen species generation, and the expression of heme oxygenase‐1 and Gadd153

“…Of these morphological alterations, cellular shrinkage, membrane blebbing and the change into round morphology, which is a typical phenomenon associated with cells undergoing either apoptosis or necrosis (Majno and Joris, 1995), were observed only at a high concentration of 4HPR (5 mM). A recent study from our laboratory has shown that relatively low concentrations of 4HPR (1 mM) induce differentiation of cultured ARPE-19 cells into a neuronal phenotype (Chen et al, 2003). Clifford et al (1999) have observed that 4HPR at high concentrations (10 mM) induces apoptosis of F9 embryonal carcinoma cells, whereas at low concentrations (1 mM) it induces differentiation of these cells into a primitive endodermal phenotype.…”

“…We have shown earlier that retinoid receptors are present in human retinal pigment epithelial cell line (ARPE-19), a cell line noted for its ability to retain many structural and functional characteristics of intact RPE, and that AGN194301, an RARa-specific antagonist, effectively blocked the RAinduced expression of stearoyl-CoA desaturase, a microsomal enzyme involved in differentiation and apoptosis (Dunn et al, 1996;Samuel et al, 2001). Interestingly, during our studies on the effect of various RA derivatives in RPE cells, we found that relatively low concentrations of 4HPR can induce differentiation of cultured ARPE-19 cells into a neuronal phenotype (Chen et al, 2003). However, these cells showed morphological changes including cell shrinkage and cell death when exposed to high concentrations of 4HPR.…”