Yu Daozhan, Swaroop Manju, Wang Mengqiao, Baxa Ulrich, Yang Rongze, Yan Yiping, Coksaygan Turhan, DeTolla Louis, Marugan Juan J, Austin Christopher P, McKew John C, Gong Da-Wei, Zheng Wei
Division of Endocrinology, Diabetes and Nutrition, Department of Medicine, University of Maryland School of Medicine, Baltimore, MD, USA.
National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MD, USA.
J Biomol Screen. 2014 Sep;19(8):1164-73. doi: 10.1177/1087057114537378. Epub 2014 Jun 6.
Niemann-Pick disease type C (NPC) is a rare neurodegenerative disorder caused by recessive mutations in the NPC1 or NPC2 gene that result in lysosomal accumulation of unesterified cholesterol in patient cells. Patient fibroblasts have been used for evaluation of compound efficacy, although neuronal degeneration is the hallmark of NPC disease. Here, we report the application of human NPC1 neural stem cells as a cell-based disease model to evaluate nine compounds that have been reported to be efficacious in the NPC1 fibroblasts and mouse models. These cells are differentiated from NPC1 induced pluripotent stem cells and exhibit a phenotype of lysosomal cholesterol accumulation. Treatment of these cells with hydroxypropyl-β-cyclodextrin, methyl-β-cyclodextrin, and δ-tocopherol significantly ameliorated the lysosomal cholesterol accumulation. Combined treatment with cyclodextrin and δ-tocopherol shows an additive or synergistic effect that otherwise requires 10-fold higher concentration of cyclodextrin alone. In addition, we found that hydroxypropyl-β-cyclodextrin is much more potent and efficacious in the NPC1 neural stem cells compared to the NPC1 fibroblasts. Miglustat, suberoylanilide hydroxamic acid, curcumin, lovastatin, pravastatin, and rapamycin did not, however, have significant effects in these cells. The results demonstrate that patient-derived NPC1 neural stem cells can be used as a model system for evaluation of drug efficacy and study of disease pathogenesis.
尼曼-匹克C型病（NPC）是一种罕见的神经退行性疾病，由NPC1或NPC2基因的隐性突变引起，导致患者细胞中未酯化胆固醇在溶酶体中蓄积。尽管神经元变性是NPC病的标志，但患者成纤维细胞已被用于评估化合物的疗效。在此，我们报告了将人NPC1神经干细胞作为一种基于细胞的疾病模型，以评估九种据报道在NPC1成纤维细胞和小鼠模型中有效的化合物。这些细胞由NPC1诱导多能干细胞分化而来，并表现出溶酶体胆固醇蓄积的表型。用羟丙基-β-环糊精、甲基-β-环糊精和δ-生育酚处理这些细胞可显著改善溶酶体胆固醇蓄积。环糊精与δ-生育酚联合处理显示出相加或协同效应，否则单独使用环糊精需要高10倍的浓度。此外，我们发现羟丙基-β-环糊精在NPC1神经干细胞中比在NPC1成纤维细胞中更有效。然而，米格鲁司他、辛二酰苯胺异羟肟酸、姜黄素、洛伐他汀、普伐他汀和雷帕霉素在这些细胞中没有显著作用。结果表明，患者来源的NPC1神经干细胞可作为评估药物疗效和研究疾病发病机制的模型系统。
