In vitro Pharmacological Evaluation in Mucopolysaccharidosis (MPS) IIIB Patient Derived Neural Stem Cells

Authors

• MG Lorino
• A Rodriguez-Lopez
• G Chen
• C Chen
• W Zheng

Abstract

Mucopolysaccharidoses (MPS) are a group of diseases in lysosomal storage disorders (LSDs), which are caused by the progressive accumulation of glycosaminoglycans (GAGs) within the lysosomes. MPS Type IIIB is a subtype caused by mutations in the NAGLU gene, which disrupts the heparan sulfate degradation pathway and causes degeneration in both somatic and central nervous systems. Additionally, due to the accumulation of GAGs, patients exhibit an enlarged lysosome phenotype as well as secondary lipid storage in the lysosomes. In this work, we have applied high throughput screening assays to evaluate the efficacy of candidate drugs on reduction of disease phenotypes in patient derived cells. To carry out this study, neural stem cells (NSCs) were derived from both MPSIIIB patient and healthy control iPSCs. The NSCs were treated with beta-cyclodextrin and delta-tocopherol, two compounds reduced accumulations of cholesterol and other lipids in cells with the Niemann Pick disease type C. After a 72-hour incubation, NSCs were stained with Lysotracker and Nile-Red dyes to assess the phenotypic changes in lysosome size and lipid accumulations, respectively. Both beta-cyclodextrin and delta-tocopherol were found to reduce enlarged lysosomes (indicated by reduced Lysotracker staining) and decrease in lipid accumulation (decreased Nile-Red staining) in the patient NSCs. Our results demonstrate the utility of this assay in high throughput screen to discover potential compounds for the treatment of MPS.

Scientific Focus Area: Stem Cell Biology

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