Induced pluripotent stem cell model recapitulates pathologic hallmarks of Gaucher disease
Induced pluripotent stem cell model recapitulates pathologic hallmarks of Gaucher disease
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DOI：
10.1073/pnas.1207889109
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发表时间：
2012-10-30
影响因子：
11.1
通讯作者：
Feldman, Ricardo A.
中科院分区：
文献类型：
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作者：
Panicker, Leelamma M.;Miller, Diana;Feldman, Ricardo A.
戈谢病（GD）是由酸性β-葡萄糖脑苷脂酶基因突变引起的常染色体隐性遗传疾病。为了对GD建模，我们通过重编程来自1型（N370 S/N370 S）、2型（L444 P/RecNP）和3型（L444 P/L444 P）GD患者的皮肤成纤维细胞来产生人诱导多能干细胞（hiPSC）。通过GD hiPSC分化为所有三个胚层并在体内形成畸胎瘤的能力证明了多能性。GD hiPSC有效地分化为在GD中受影响最大的细胞类型，即，巨噬细胞和神经细胞。GD hiPSC-巨噬细胞表达巨噬细胞特异性标志物，具有吞噬作用，并且能够响应LPS释放炎性介质。此外，GD hiPSC-巨噬细胞概括了该疾病的表型特征。他们表现出低葡萄糖脑苷脂酶（GC）的酶活性和积累鞘脂，其溶酶体功能严重受损。GD hiPSC-巨噬细胞在清除吞噬的RBC（组织浸润GD巨噬细胞的表型）的能力方面存在缺陷。1型、2型和3型GD hiPSC-巨噬细胞的RBC清除动力学与突变的严重程度相关。与重组GC一起孵育完全逆转了所有三种类型的GD hiPSC-巨噬细胞的RBC清除延迟，这表明它们的功能缺陷确实是由GC缺陷引起的。然而，治疗诱导巨噬细胞与伴侣isofagomine恢复吞噬的RBC清除只有部分，无论基因型。这些发现与重组GC和异法戈明的已知临床疗效一致。我们得出结论，源自GD hiPSC的细胞类型可以有效地概括该疾病的病理学特征。
Gaucher disease (GD) is an autosomal recessive disorder caused by mutations in the acid beta-glucocerebrosidase gene. To model GD, we generated human induced pluripotent stem cells (hiPSC), by reprogramming skin fibroblasts from patients with type 1 (N370S/N370S), type 2 (L444P/RecNcil), and type 3 (L444P/L444P) GD. Pluripotency was demonstrated by the ability of GD hiPSC to differentiate to all three germ layers and to form teratomas in vivo. GD hiPSC differentiated efficiently to the cell types most affected in GD, i.e., macrophages and neuronal cells. GD hiPSC-macrophages expressed macrophage-specific markers, were phagocytic, and were capable of releasing inflammatory mediators in response to LPS. Moreover, GD hiPSC-macrophages recapitulated the phenotypic hallmarks of the disease. They exhibited low glucocerebrosidase (GC) enzymatic activity and accumulated sphingolipids, and their lysosomal functions were severely compromised. GD hiPSC-macrophages had a defect in their ability to clear phagocytosed RBC, a phenotype of tissue-infiltrating GD macrophages. The kinetics of RBC clearance by types 1, 2, and 3 GD hiPSC-macrophages correlated with the severity of the mutations. Incubation with recombinant GC completely reversed the delay in RBC clearance from all three types of GD hiPSC-macrophages, indicating that their functional defects were indeed caused by GC deficiency. However, treatment of induced macrophages with the chaperone isofagomine restored phagocytosed RBC clearance only partially, regardless of genotype. These findings are consistent with the known clinical efficacies of recombinant GC and isofagomine. We conclude that cell types derived from GD hiPSC can effectively recapitulate pathologic hallmarks of the disease.