Department of Biology and Biotechnology "C. Darwin", Sapienza University of Rome, 00185 Rome, Italy.
Department of Molecular Medicine, Sapienza University of Rome, 00161 Rome, Italy.
Int J Mol Sci. 2018 Jun 20;19(6):1814. doi: 10.3390/ijms19061814.
The term orthodisease defines human disorders in which the pathogenic gene has orthologs in model organism genomes. Yeasts have been instrumental for gaining insights into the molecular basis of many human disorders, particularly those resulting from impaired cellular metabolism. We and others have used yeasts as a model system to study the molecular basis of Hailey-Hailey disease (HHD), a human blistering skin disorder caused by haploinsufficiency of the gene the orthologous of the yeast gene . We observed that cells defective for gene share several biological similarities with HHD derived keratinocytes. Based on the conservation of ATP2C1/PMR1 function from yeast to human, here we used a yeast-based assay to screen for molecules able to influence the pleiotropy associated with deletion. We identified six compounds, Kaempferol, Indirubin, Lappaconite, Cyclocytidine, Azomycin and Nalidixic Acid that induced different major shape phenotypes in . These include mitochondrial and the cell-wall morphology-related phenotypes. Interestingly, a secondary assay in mammalian cells confirmed activity for Kaempferol. Indeed, this compound was also active on human keratinocytes depleted of ATP2C1 function by siRNA-treatment used as an in-vitro model of HHD. We found that Kaempferol was a potent NRF2 regulator, strongly inducing its expression and its downstream target . In addition, Kaempferol could decrease oxidative stress of ATP2C1 defective keratinocytes, characterized by reduced NRF2-expression. Our results indicated that the activation of these pathways might provide protection to the HHD-skin cells. As oxidative stress plays pivotal roles in promoting the skin lesions of Hailey-Hailey, the NRF2 pathway could be a viable therapeutic target for HHD.
术语“orthodisease”定义了人类疾病，其中致病基因在模式生物基因组中有同源物。酵母在深入了解许多人类疾病的分子基础方面发挥了重要作用，特别是那些由于细胞代谢受损而导致的疾病。我们和其他人已经使用酵母作为模型系统来研究 Hailey-Hailey 病（HHD）的分子基础，HHD 是一种人类水疱性皮肤病，由基因的单倍不足引起，该基因的同源物在酵母中是 。我们观察到，基因缺陷的细胞与源自 HHD 的角质形成细胞具有几种生物学相似性。基于酵母到人类中 ATP2C1/PMR1 功能的保守性，我们在这里使用基于酵母的测定法来筛选能够影响与缺失相关的表型多样性的分子。我们鉴定出六种化合物，即山萘酚、靛玉红、紫菀酮、环胞苷、阿佐霉素和萘啶酸，它们在 中诱导不同的主要形态表型。这些包括线粒体和细胞壁形态相关的表型。有趣的是，在哺乳动物细胞中的二次测定证实了山萘酚的活性。事实上，这种化合物对通过 siRNA 处理耗尽 ATP2C1 功能的人角质形成细胞也具有活性，siRNA 处理被用作 HHD 的体外模型。我们发现山萘酚是一种有效的 NRF2 调节剂，强烈诱导其表达及其下游靶标 。此外，山萘酚可以降低 ATP2C1 缺陷角质形成细胞的氧化应激，其特征是 NRF2 表达减少。我们的结果表明，这些途径的激活可能为 HHD 皮肤细胞提供保护。由于氧化应激在促进 Hailey-Hailey 的皮肤损伤中起着关键作用，因此 NRF2 途径可能是 HHD 的可行治疗靶点。
