Identifying drug targets for Niemann-Pick type C1 disease

Authors

• P Massa
• E Chiang
• R Luke Y Chen
• N Cawley
• D Porter
• K Cheng

Abstract

Niemann-Pick type C1 (NPC1) disease is a neurodegenerative disorder caused by the accumulation of endolysosomal unesterified cholesterol in cells that are deficient in NPC intracellular cholesterol transporter 1 (NPC1) [1]. Our research aims to identify small molecules with therapeutic potential for NPC1 in iPSC-derived neurons. Previous studies were focused on NPC1-deficient fibroblasts. However, NPC1 is fundamentally a neurological disease. It is critical to identify and understand the cellular processes in a physiologically relevant model. In this study, we utilized NPC1 -/- i3Neurons that can be rapidly differentiated into glutamatergic neurons in a scalable, two-step process yielding homogenous neuronal cultures 10 days post-induction. NPC1 -/- neurons were treated with either 2-hydroxypropyl-B-cyclodextrin (HPBCD) or mock and compared with wild-type (WT) neurons. HPBCD is a compound already known to reduce unesterified cholesterol accumulation and will be used as a positive control for the screen to come [2]. Unesterified cholesterol buildup was visualized by staining with the fluorescently labeled cholesterol-binding agent perfringolysin-O (PFO). The assay condition is optimized in 384 plate format which is amenable for the high-throughput small molecule screen. We reported NPC1 -/- i3Neurons treated with HPBCD showed a statistically significant reduction in endolysosomal unesterified cholesterol, suggesting that it will serve as a benchmark to evaluate the efficacy of small molecules in the drug library.

Scientific Focus Area: Neuroscience

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