PDF(Pubmed)
Abstract：
UNASSIGNED: Autosomal recessive spastic ataxia of Charlevoix-Saguenay (ARSACS) is a rare incurable neurodegenerative disease caused by mutations in the SACS gene, which codes for sacsin, a large protein involved in protein homeostasis, mitochondrial function, cytoskeletal dynamics, autophagy, cell adhesion and vesicle trafficking. However, the pathogenic mechanisms underlying sacsin dysfunction are still largely uncharacterized, and so attempts to develop therapies are still in the early stages.
UNASSIGNED: To achieve further understanding of how processes are altered by loss of sacsin, we used untargeted proteomics to compare protein profiles in ARSACS fibroblasts versus controls.
UNASSIGNED: Our analyses confirmed the involvement of known biological pathways and also implicated calcium and lipid homeostasis in ARSACS skin fibroblasts, a finding further verified in SH-SY5Y SACS -/- cells. Validation through mass spectrometry-based analysis and comparative quantification of lipids by LC-MS in fibroblasts revealed increased levels of ceramides coupled with a reduction of diacylglycerols.
UNASSIGNED: In addition to confirming aberrant Ca2+ homeostasis in ARSACS, this study described abnormal lipid levels associated with loss of sacsin.
UNASSIGNED: To achieve further understanding of how processes are altered by loss of sacsin, we used untargeted proteomics to compare protein profiles in ARSACS fibroblasts versus controls.
UNASSIGNED: Our analyses confirmed the involvement of known biological pathways and also implicated calcium and lipid homeostasis in ARSACS skin fibroblasts, a finding further verified in SH-SY5Y SACS -/- cells. Validation through mass spectrometry-based analysis and comparative quantification of lipids by LC-MS in fibroblasts revealed increased levels of ceramides coupled with a reduction of diacylglycerols.
UNASSIGNED: In addition to confirming aberrant Ca2+ homeostasis in ARSACS, this study described abnormal lipid levels associated with loss of sacsin.
摘要：
■Charlevoix-Saguenay常染色体隐性遗传性痉挛性共济失调（ARSACS）是由SACS基因突变引起的一种罕见的无法治愈的神经退行性疾病，萨辛的代码，一种参与蛋白质稳态的大蛋白质，线粒体功能，细胞骨架动力学，自噬,细胞粘附和囊泡运输。然而,sacsin功能障碍的致病机制仍然在很大程度上没有被描述，因此，开发疗法的尝试仍处于早期阶段。
■为了进一步了解sacsin的丢失是如何改变过程的，我们使用非靶向蛋白质组学比较了ARSACS成纤维细胞与对照的蛋白质谱.
■我们的分析证实了ARSACS皮肤成纤维细胞中已知的生物学途径以及钙和脂质稳态的参与，这一发现在SH-SY5YSACS-/-细胞中得到了进一步验证。通过基于质谱的分析和通过LC-MS对成纤维细胞中的脂质进行比较定量的验证表明，神经酰胺的水平增加，二酰甘油的减少。
■除了证实ARSACS中异常的Ca2+稳态外，这项研究描述了与sacsin丢失相关的异常脂质水平。
■为了进一步了解sacsin的丢失是如何改变过程的，我们使用非靶向蛋白质组学比较了ARSACS成纤维细胞与对照的蛋白质谱.
■我们的分析证实了ARSACS皮肤成纤维细胞中已知的生物学途径以及钙和脂质稳态的参与，这一发现在SH-SY5YSACS-/-细胞中得到了进一步验证。通过基于质谱的分析和通过LC-MS对成纤维细胞中的脂质进行比较定量的验证表明，神经酰胺的水平增加，二酰甘油的减少。
■除了证实ARSACS中异常的Ca2+稳态外，这项研究描述了与sacsin丢失相关的异常脂质水平。
