A Novel Role of ARA70 in Regulating Ferritinophagy of RGCs During Retinal Ischemia Reperfusion.

Abstract

Although the contribution of ferroptosis, an iron-dependent cell death, to ischemia reperfusion (IR)-induced retinal injury has been reported before, to optimize therapeutic strategy, there is still an urgent need to identify potential candidates involved in this process. Androgen Receptor-Associated Protein of 70 kDa (ARA70) is a cargo receptor for ferritinophagy, and its role in retinal ferroptosis has not been revealed yet. Herein, we explored the role of ARA70 in IR-associated retinal lesions by in vivo (C57BL/6 J mice with intraocular pressure of 90-100 mmHg) and in vitro (retinal ganglion cells (RGCs) stimulated with tert-butyl hydroperoxide (tBH)) experiments. It was found that IR upregulated ARA70 expression and accelerated lipid peroxidation in retinal tissues. We first confirmed that two ferroptosis inhibitors, deferiprone or ferrostatin-1 (Fer-1), suppressed ferritin degradation, restrained apoptosis and inflammation, and protected mouse retinas against IR stress. Next, primary mouse RGCs were treated with tBH to simulate IR environment in vitro. ARA70 expression was decreased at lower concentrations of tBH (5-20 μM), but increased at higher concentrations (40-80 μM). Interestingly, the expression of ferritin-related proteins (ferritin heavy chain, FTH; ferritin light chain, FTL) showed an opposite alteration. Knockdown of ARA70 protected RGCs from tBH-induced damage. It inhibited the delivery of ferritin to lysosomes for ferritinophagy and thus reducing cellular Fe²⁺ concentration. Besides, ARA70 knockdown suppressed autophagy and inflammation of tBH-treated RGCs. These findings provide novel insights into the pathogenesis of retinal IR, and may be helpful for treatment of retinal diseases.