Cholesterol enhances amyloid β deposition in mouse retina by modulating the activities of Aβ-regulating enzymes in retinal pigment epithelial cells

Subretinally-deposited amyloid β (Aβ) is a main contributor of developing age-related macular degeneration (AMD). However, the mechanism causing Aβ deposition in AMD eyes is unknown. Hypercholesterolemia is a significant risk for developing AMD. Thus, we investigated the effects of cholesterol on Aβ production in retinal pigment epithelial (RPE) cells in vitro and in the mouse retina in vivo. RPE cells isolated from senescent (12-month-old) C57BL/6 mice were treated with 10 μg/ml cholesterol for 48 h. Aβ amounts in culture supernatants were measured by ELISA. Activity and expression of enzymes and proteins that regulate Aβ production were examined by activity assay and real time PCR. The retina of mice fed cholesterol-enriched diet was examined by transmission electron microscopy. Cholesterol significantly increased Aβ production in cultured RPE cells. Activities of Aβ degradation enzyme; neprilysin (NEP) and anti-amyloidogenic secretase; α-secretase were significantly decreased in cell lysates of cholesterol-treated RPE cells compared to non-treated cells, but there was no change in the activities of β- or γ-secretase. mRNA levels of NEP and α-secretase (ADAM10 and ADAM17) were significantly lower in cholesterol-treated RPE cells than non-treated cells. Senescent (12-month-old) mice fed cholesterol-enriched chow developed subRPE deposits containing Aβ, whereas age-matched mice fed standard rodent chow diet did not. Activities and mRNA levels of NEP and α-secretase were significantly lower in native RPE cells freshly isolated from cholesterol-enriched chow fed mice compared to standard rodent chow fed mice. These findings suggest that cholesterol enhances subretinal Aβ accumulation by modulating the activities of enzymes degrading and processing Aβ in RPE cells in senescent subjects.

► Cholesterol-treated RPE produces more Aβ than non-treated RPE.
► Neprilysin expression and activity decreased in cholesterol-treated RPE.
► α-Secretase expression and activity decreased in cholesterol-treated RPE.
► Cholesterol-enriched diet induced subRPE deposits in aged mice.
► Aβ were present in cholesterol-enriched-diet-induced subRPE deposits in aged mice.