National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MD 20892, USA.
iPSC Core Facility, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA.
Cells. 2020 Dec 22;10(1):8. doi: 10.3390/cells10010008.
Pompe disease is a lysosomal storage disorder caused by autosomal recessive mutations in the gene. Acid alpha-glucosidase deficiency leads to abnormal glycogen accumulation in patient cells. Given the increasing evidence of central nervous system (CNS) involvement in classic infantile Pompe disease, we used neural stem cells, differentiated from patient induced pluripotent stem cells, to model the neuronal phenotype of Pompe disease. These Pompe neural stem cells exhibited disease-related phenotypes including glycogen accumulation, increased lysosomal staining, and secondary lipid buildup. These morphological phenotypes in patient neural stem cells provided a tool for drug efficacy evaluation. Two potential therapeutic agents, hydroxypropyl-β-cyclodextrin and δ-tocopherol, were tested along with recombinant human acid alpha-glucosidase (rhGAA) in this cell-based Pompe model. Treatment with rhGAA reduced LysoTracker staining in Pompe neural stem cells, indicating reduced lysosome size. Additionally, treatment of diseased neural stem cells with the combination of hydroxypropyl-β-cyclodextrin and δ-tocopherol significantly reduced the disease phenotypes. These results demonstrated patient-derived Pompe neural stem cells could be used as a model to study disease pathogenesis, to evaluate drug efficacy, and to screen compounds for drug discovery in the context of correcting CNS defects.
庞贝病是一种溶酶体贮积症，由 基因的常染色体隐性突变引起。酸性α-葡萄糖苷酶缺乏导致患者细胞中异常的糖原积累。鉴于经典婴儿型庞贝病中枢神经系统（CNS）受累的证据不断增加，我们使用神经干细胞，从患者诱导多能干细胞分化而来，来模拟庞贝病的神经元表型。这些庞贝病神经干细胞表现出与疾病相关的表型，包括糖原积累、溶酶体染色增加和继发性脂质堆积。患者神经干细胞中的这些形态表型为药物疗效评估提供了工具。在这个基于细胞的庞贝病模型中，我们测试了两种潜在的治疗剂，羟丙基-β-环糊精和 δ-生育酚，以及重组人酸性α-葡萄糖苷酶（rhGAA）。rhGAA 的治疗降低了庞贝病神经干细胞中的 LysoTracker 染色，表明溶酶体体积减小。此外，用羟丙基-β-环糊精和 δ-生育酚联合治疗患病的神经干细胞，显著降低了疾病表型。这些结果表明，源自患者的庞贝病神经干细胞可用于研究疾病发病机制、评估药物疗效，并在纠正中枢神经系统缺陷的背景下筛选化合物以进行药物发现。
