Synthetic mRNA-based differentiation method enables early detection of Parkinson's phenotypes in neurons derived from Gaucher disease-induced pluripotent stem cells.
Synthetic mRNA-based differentiation method enables early detection of Parkinson's phenotypes in neurons derived from Gaucher disease-induced pluripotent stem cells.
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DOI：
10.1002/sctm.20-0302
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发表时间：
2021-04
影响因子：
6
通讯作者：
Iwata H
中科院分区：
文献类型：
--
作者：
Akiyama T;Sato S;Ko SBH;Sano O;Sato S;Saito M;Nagai H;Ko MSH;Iwata H
高谢病是最常见的代谢性储存障碍，由葡萄糖脑苷酶基因GBA1突变引起，导致葡萄糖神经酰胺(GlcCer)在受影响的细胞中积聚。高谢病1型(GD1)，虽然被定义为非神经病变的亚型，但伴随着帕金森氏病风险的增加。为了深入了解GlcCer进行性积聚与帕金森病表型的关系，我们从一名GD1患者和一名健康捐赠者对照的诱导多能干细胞(IPSCs)中产生多巴胺(DA)神经元，并用液相色谱-质谱仪测量GlcCer积聚。我们测试了两种DA神经元分化方法：一种成熟的方法模拟了从IPSCs到神经前体细胞和DA神经元的循序渐进的发育过程；另一种是基于合成mRNA的方法，在IPSCs中过度表达转录因子。前者在分化60 后可检测到Gd1特异性的GlcCer积累，而后者仅在10 d后检测到GlcCer。利用这种基于合成mRNA的快速分化方法，我们发现GD1患者细胞中的代谢缺陷可以通过野生型GBA1的过表达或GlcCer合成抑制剂的处理来挽救。此外，我们还检测到帕金森氏病的生物标志物α-突触核蛋白在一例患者的DA神经元中的磷酸化增加，而野生型GBA1的过度表达显著降低了这种磷酸化。这些结果表明，基于合成信使核糖核酸的方法加速了对GD1患者帕金森病病理机制的分析，并可能促进药物发现过程。转录因子的合成表达增强了诱导的多能干细胞分化，并使早期(10 天)能够检测到帕金森病的风险因素，即葡萄糖神经酰胺(GlcCer)在高雪病患者神经元中的积累。这种快速的鉴别方法也使得能够检测到α-突触核蛋白的磷酸化--一种与帕金森氏病相关的关键表型。
Gaucher disease, the most prevalent metabolic storage disorder, is caused by mutations in the glucocerebrosidase gene GBA1, which lead to the accumulation of glucosylceramide (GlcCer) in affected cells. Gaucher disease type 1 (GD1), although defined as a nonneuronopathic subtype, is accompanied by an increased risk of Parkinson's disease. To gain insights into the association of progressive accumulation of GlcCer and the Parkinson's disease phenotypes, we generated dopaminergic (DA) neurons from induced pluripotent stem cells (iPSCs) derived from a GD1 patient and a healthy donor control, and measured GlcCer accumulation by liquid chromatography‐mass spectrometry. We tested two DA neuron differentiation methods: a well‐established method that mimics a step‐wise developmental process from iPSCs to neural progenitor cells, and to DA neurons; and a synthetic mRNA‐based method that overexpresses a transcription factor in iPSCs. GD1‐specific accumulation of GlcCer was detected after 60 days of differentiation by the former method, whereas it was detected after only 10 days by the latter method. With this synthetic mRNA‐based rapid differentiation method, we found that the metabolic defect in GD1 patient cells can be rescued by the overexpression of wild‐type GBA1 or the treatment with an inhibitor for GlcCer synthesis. Furthermore, we detected the increased phosphorylation of α‐synuclein, a biomarker for Parkinson's disease, in DA neurons derived from a GD1 patient, which was significantly decreased by the overexpression of wild‐type GBA1. These results suggest that synthetic mRNA‐based method accelerates the analyses of the pathological mechanisms of Parkinson's disease in GD1 patients and possibly facilitates drug discovery processes. Synthetic mRNA‐based expression of transcription factors enhanced induced pluripotent stem cell differentiation and enabled early (10 days) detection of a risk factor of Parkinson's disease, i.e. the accumulation of glucosylceramide (GlcCer) in Gaucher patient neurons. This rapid differentiation method also enabled to detect the phosphorylation of α‐synuclein—a critical Parkinson's disease‐linked phenotype.
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影响因子：
64.8
作者：
Kriks, Sonja;Shim, Jae-Won;Piao, Jinghua;Ganat, Yosif M.;Wakeman, Dustin R.;Xie, Zhong;Carrillo-Reid, Luis;Auyeung, Gordon;Antonacci, Chris;Buch, Amanda;Yang, Lichuan;Beal, M. Flint;Surmeier, D. James;Kordower, Jeffrey H.;Tabar, Viviane;Studer, Lorenz
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通讯作者：
Mistry, Pramod K.
影响因子：
21.3
作者：
Fujiwara, H;Hasegawa, M;Iwatsubo, T
通讯作者：
Iwatsubo, T