Voccoli V, Tonazzini I, Signore G, Caleo M, Cecchini M
NEST, Istituto Nanoscienze-CNR and Scuola Normale Superiore, Piazza San Silvestro 12, 56127 Pisa, Italy.
Center for Nanotechnology Innovation@NEST, Istituto Italiano di Tecnologia, Pisa, Italy.
Cell Death Dis. 2014 Nov 20;5(11):e1529. doi: 10.1038/cddis.2014.483.
Globoid cell leukodystrophy (GLD) is a metabolic disease caused by mutations in the galactocerebrosidase (GALC) gene. GALC is a lysosomal enzyme whose function is to degrade galacto-lipids, including galactosyl-ceramide and galactosyl-sphingosine (psychosine, PSY). GALC loss of function causes progressive intracellular accumulation of PSY. It is widely held that PSY is the main trigger for the degeneration of myelinating cells and progressive white-matter loss. However, still little is known about the molecular mechanisms by which PSY imparts toxicity. Here, we address the role of calcium dynamics during PSY-induced cell death. Using the human oligodendrocyte cell line MO3.13, we report that cell death by PSY is accompanied by robust cytosolic and mitochondrial calcium (Ca(2+)) elevations, and by mitochondrial reactive oxygen species (ROS) production. Importantly, we demonstrate that the reduction of extracellular calcium content by the chelating agent ethylenediaminetetraacetic acid can decrease intra-mitochondrial ROS production and enhance cell viability. Antioxidant administration also reduces mitochondrial ROS production and cell loss, but this treatment does not synergize with Ca(2+) chelation. Our results disclose novel intracellular pathways involved in PSY-induced death that may be exploited for therapeutic purposes to delay GLD onset and/or slow down its progression.
球形细胞脑白质营养不良症(GLD)是一种由半乳糖脑苷脂酶(GALC)基因突变引起的代谢性疾病。GALC是一种溶酶体酶,其功能是降解半乳糖脂类,包括半乳糖神经酰胺和半乳糖神经鞘氨醇(鞘氨醇半乳糖苷,PSY)。GALC功能丧失会导致PSY在细胞内逐渐积累。人们普遍认为,PSY是导致髓鞘形成细胞变性和进行性白质损失的主要诱因。然而,关于PSY产生毒性的分子机制仍知之甚少。在此,我们探讨了钙动力学在PSY诱导的细胞死亡过程中的作用。利用人少突胶质细胞系MO3.13,我们发现PSY诱导的细胞死亡伴随着胞质和线粒体钙(Ca(2+))的显著升高以及线粒体活性氧(ROS)的产生。重要的是,我们证明螯合剂乙二胺四乙酸降低细胞外钙含量可减少线粒体内ROS的产生并提高细胞活力。给予抗氧化剂也可减少线粒体ROS的产生和细胞损失,但这种处理与Ca(2+)螯合并无协同作用。我们的研究结果揭示了PSY诱导细胞死亡所涉及的新的细胞内途径,这些途径可能被用于治疗目的,以延缓GLD的发病和/或减缓其进展。