iPSC-derived neurons from GBA1-associated Parkinson's disease patients show autophagic defects and impaired calcium homeostasis
iPSC-derived neurons from GBA1-associated Parkinson's disease patients show autophagic defects and impaired calcium homeostasis
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DOI：
10.1038/ncomms5028
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发表时间：
2014-06-01
影响因子：
16.6
通讯作者：
Deleidi, Michela
中科院分区：
文献类型：
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作者：
Schoendorf, David C.;Aureli, Massimo;Deleidi, Michela
酸性β-葡萄糖脑苷脂酶（GBA 1）基因的突变导致溶酶体贮积症戈谢病（GD），是迄今已知的帕金森病（PD）最强的遗传风险因素。在这里，我们从携带GBA 1突变的GD和PD受试者中产生诱导多能干细胞，并将其分化为中脑多巴胺能神经元，然后使用荧光激活细胞分选进行富集。神经元显示葡萄糖脑苷脂酶活性和蛋白质水平降低，葡萄糖神经酰胺和α-突触核蛋白水平增加以及自噬和溶酶体缺陷。定量蛋白质组分析揭示了患病神经元中神经元钙结合蛋白2（NECAB 2）的增加。突变的神经元表现出钙稳态失调和增加的脆弱性，涉及胞质钙升高的应激反应。重要的是，突变的校正挽救了这种病理表型。这些发现为GBA 1突变与自噬/溶酶体系统和细胞内钙稳态的复杂变化之间的联系提供了证据，这是神经变性的脆弱性的基础。
Mutations in the acid b-glucocerebrosidase (GBA1) gene, responsible for the lysosomal storage disorder Gaucher's disease (GD), are the strongest genetic risk factor for Parkinson's disease (PD) known to date. Here we generate induced pluripotent stem cells from subjects with GD and PD harbouring GBA1 mutations, and differentiate them into midbrain dopaminergic neurons followed by enrichment using fluorescence-activated cell sorting. Neurons show a reduction in glucocerebrosidase activity and protein levels, increase in glucosylceramide and a-synuclein levels as well as autophagic and lysosomal defects. Quantitative proteomic profiling reveals an increase of the neuronal calcium-binding protein 2 (NECAB2) in diseased neurons. Mutant neurons show a dysregulation of calcium homeostasis and increased vulnerability to stress responses involving elevation of cytosolic calcium. Importantly, correction of the mutations rescues such pathological phenotypes. These findings provide evidence for a link between GBA1 mutations and complex changes in the autophagic/lysosomal system and intracellular calcium homeostasis, which underlie vulnerability to neurodegeneration.