Induced pluripotent stem cell model recapitulates pathologic hallmarks of Gaucher disease

Abstract

Gaucher disease (GD) is an autosomal recessive disorder caused by mutations in the acid β-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/RecNciI), 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.Gaucher model | Gaucher macrophages | lipid storage disease | glucosylsphingolipids | erythrophagocytosis

Macrophage Models of Gaucher Disease for Evaluating Disease Pathogenesis and Candidate Drugs

“…We generated and studied primary macrophages (hMacs) that were differentiated from monocytes taken from patients with Gaucher disease. However, the amount of blood needed, especially from infants with Gaucher disease type 2, and the limited yield of macrophages prompted us to create induced pluripotent stem cells (iPSCs) from dermal fibroblasts taken from patients to generate and evaluate a clonal supply of macrophages (iMacs) (13). Here, we use hMacs and iMacs to evaluate the ability of the noninhibitory chaperone molecule NCGC00188758 to reverse the disease phenotype.…”

Stem Cells and Neuronal Differentiation

“…Neural crest progenitors were also derived from iPSCs and compared with those from hESCs, followed by differentiation to functional Schwann cells [104]. More than the therapeutic use of neural progenitors derived from hiPSCs, this in vitro cell-based technology is in the spotlight to reproduce cellular models of poorly understood diseases -such as Down syndrome, Friedreich's ataxia, Gaucher disease [105][106][107], amyotrophic lateral sclerosis [108], spinal muscular atrophy [109], Parkinson's disease [110,111], schizophrenia [112], Huntington's disease [110], and Alzheimer's disease [113]. Furthermore, correction of genetic mutations in disease-specific iPS cells can rescue phenotypes in cultured cells [111,114] or in mouse models of human diseases, such as sickle cell anemia [115].…”

New Macrophage Models of Gaucher Disease Offer New Tools for Drug Development

“…They also demonstrated defective phagocytosis in these iPSC-derived GMs [11] , and more recently showed that these cells exhibit greatly increased secretion of the pro-inflammatory cytokines tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6 in response to lipopolysaccharide (LPS) treatment [12] . These results reflect the increased expression of these cytokines observed in some patients with type 1 GD [2,4] .…”

“…Panicker et al derived iPSC lines from patients with all three forms of GD [11,12] . They also demonstrated defective phagocytosis in these iPSC-derived GMs [11] , and more recently showed that these cells exhibit greatly increased secretion of the pro-inflammatory cytokines tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6 in response to lipopolysaccharide (LPS) treatment [12] .…”