Expression of ABCA4 in the retinal pigment epithelium and its implications for Stargardt macular degeneration.

Abstract

Recessive Stargardt disease (STGD1) is an inherited blinding disorder caused by mutations in the <i>Abca4</i> gene. ABCA4 is a flippase in photoreceptor outer segments (OS) that translocates retinaldehyde conjugated to phosphatidylethanolamine across OS disc membranes. Loss of ABCA4 in <i>Abca4</i>^<i>-/-</i> mice and STGD1 patients causes buildup of lipofuscin in the retinal pigment epithelium (RPE) and degeneration of photoreceptors, leading to blindness. No effective treatment currently exists for STGD1. Here we show by several approaches that ABCA4 is additionally expressed in RPE cells. (<i>i</i>) By in situ hybridization analysis and by RNA-sequencing analysis, we show the <i>Abca4</i> mRNA is expressed in human and mouse RPE cells. (<i>ii</i>) By quantitative immunoblotting, we show that the level of ABCA4 protein in homogenates of wild-type mouse RPE is about 1% of the level in neural retina homogenates. (<i>iii</i>) ABCA4 immunofluorescence is present in RPE cells of wild-type and <i>Mertk</i>^<i>-/-</i> but not <i>Abca4</i>^<i>-/-</i> mouse retina sections, where it colocalizes with endolysosomal proteins. To elucidate the role of ABCA4 in RPE cells, we generated a line of genetically modified mice that express ABCA4 in RPE cells but not in photoreceptors. Mice from this line on the <i>Abca4</i>^<i>-/-</i> background showed partial rescue of photoreceptor degeneration and decreased lipofuscin accumulation compared with nontransgenic <i>Abca4</i>^<i>-/-</i> mice. We propose that ABCA4 functions to recycle retinaldehyde released during proteolysis of rhodopsin in RPE endolysosomes following daily phagocytosis of distal photoreceptor OS. ABCA4 deficiency in the RPE may play a role in the pathogenesis of STGD1.