Spincemaille Pieter, Chandhok Gursimran, Zibert Andree, Schmidt Hartmut, Verbeek Jef, Chaltin Patrick, Cammue Bruno P, Cassiman David, Thevissen Karin
Centre of Microbial and Plant Genetics (CMPG), KU Leuven, Kasteelpark Arenberg 20, 3001 Heverlee, Belgium.
Clinic for Transplantation Medicine, Münster University Hospital, Albert-Schweitzer-Campus 1, Building A14, D-48149 Münster, Germany.
Microb Cell. 2014 Oct 24;1(11):352-364. doi: 10.15698/mic2014.11.175.
The human pathology Wilson disease (WD) is characterized by toxic copper (Cu) accumulation in brain and liver, resulting in, among other indications, mitochondrial dysfunction and apoptosis of hepatocytes. In an effort to identify novel compounds that can alleviate Cu-induced toxicity, we screened the Pharmakon 1600 repositioning library using a Cu-toxicity yeast screen. We identified 2 members of the drug class of Angiotensin II Type 1 receptor blockers (ARBs) that could increase yeast tolerance to Cu, namely Candesartan and Losartan. Subsequently, we show that specific ARBs can increase yeast tolerance to Cu and/or the chemotherapeutic agent cisplatin (Cp). The latter also induces mitochondrial dysfunction and apoptosis in mammalian cells. We further demonstrate that specific ARBs can prevent the prevalence of Cu-induced apoptotic markers in yeast, with Candesartan Cilexetil being the ARB which demonstrated most pronounced reduction of apoptosis-related markers. Next, we tested the sensitivity of a selection of yeast knockout mutants affected in detoxification of reactive oxygen species (ROS) and Cu for Candesartan Cilexetil rescue in presence of Cu. These data indicate that Candesartan Cilexetil increases yeast tolerance to Cu irrespectively of major ROS-detoxifying proteins. Finally, we show that specific ARBs can increase mammalian cell tolerance to Cu, as well as decrease the prevalence of Cu-induced apoptotic markers. All the above point to the potential of ARBs in preventing Cu-induced toxicity in yeast and mammalian cells.
人类病理学中的威尔逊病（WD）的特征是大脑和肝脏中有毒铜（Cu）的积累，除其他表现外，还会导致线粒体功能障碍和肝细胞凋亡。为了鉴定能够减轻铜诱导毒性的新型化合物，我们使用铜毒性酵母筛选法对药物再定位文库Pharmakon 1600进行了筛选。我们鉴定出血管紧张素II 1型受体阻滞剂（ARB）药物类别的2个成员，即坎地沙坦和氯沙坦，它们可以提高酵母对铜的耐受性。随后，我们表明特定的ARB可以提高酵母对铜和/或化疗药物顺铂（Cp）的耐受性。后者也会诱导哺乳动物细胞中的线粒体功能障碍和凋亡。我们进一步证明特定的ARB可以防止酵母中铜诱导的凋亡标志物的出现，其中坎地沙坦酯是表现出与凋亡相关标志物最明显减少的ARB。接下来，我们测试了在铜存在的情况下，受活性氧（ROS）和铜解毒影响的一系列酵母敲除突变体对坎地沙坦酯拯救的敏感性。这些数据表明，坎地沙坦酯提高酵母对铜的耐受性与主要的ROS解毒蛋白无关。最后，我们表明特定的ARB可以提高哺乳动物细胞对铜的耐受性，并降低铜诱导的凋亡标志物的出现率。以上所有都表明ARB在预防酵母和哺乳动物细胞中铜诱导的毒性方面具有潜力。
