High-throughput assay development for Niemann-Pick type A small molecule therapeutics

Authors

• K Gorshkov
• M Xu
• S Yang
• W Zheng

Abstract

Niemann-Pick type A (NPA) is a rare lysosomal storage disease that, together with Niemann-Pick type B, affects 1 in 250,000. NPA and NPB is caused by a deficiency in the production of acid-sphingomyelinase (ASM) that is encoded by the SMPD1 gene. The early onset of NPA results in the accumulation of intracellular sphingomyelin (SM), severe neurological dysfunction, and death by four years of age. While enzyme replacement therapy is in ongoing clinical trials, novel small molecule therapeutics must also be pursued. At the NCATS TRND program, we are working on developing novel assays for high-throughput screening of FDA-approved and other drug libraries to identify compounds that may restore ASM function and alleviate the SM accumulation phenotype. To do this, our lab has generated patient cell models using NPA patient-derived fibroblasts and patient hiPSC-derived neural stem cells. Using a dye-conjugated BODIPY-SM, we are able to visualize significant accumulation of SM when comparing disease to wild-type cells using various cell-based imaging platforms. Simultaneous use of BODIPY-SM and the LysoTracker Deep Red lysosomal marker creates a robust imaging assay to verify compound efficacy. We are now developing an innovative approach with a washless, plate-based cytometer assay that will enable easy and efficient screening of small molecules to combat NPA and other lysosomal storage diseases. Altogether, these efforts will bring us closer to providing patients with desperately needed therapeutic solutions.

Scientific Focus Area: Cell Biology

This page was last updated on Friday, March 26, 2021