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A phenotypic compound screening assay for lysosomal storage diseases.
Miao Xu1, Ke Liu, Manju Swaroop
11Therapeutics for Rare and Neglected Disease, National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MD, USA.
Journal of Biomolecular Screening
|August 29, 2013
Summary
Researchers developed a novel Lysotracker assay to measure enlarged lysosomes in patient cells. This assay aids in screening compounds for lysosomal storage disease drug development.
Area of Science:
- Cell Biology
- Genetics
- Pharmacology
Background:
- Lysosomes are crucial cellular recycling centers.
- Lysosomal storage diseases result from genetic defects causing macromolecule accumulation and enlarged lysosomes.
- Lysosomes are emerging therapeutic targets for treating these diseases.
Purpose of the Study:
- To develop a suitable assay for compound screening against diseased lysosomes.
- To establish a phenotypic assay for measuring lysosome size in patient-derived cells.
Main Methods:
- Developed a Lysotracker staining assay.
- Measured enlarged lysosomes using fluorescence intensity and microscopic analysis.
- Tested the assay in patient cells from various lysosomal storage diseases.
Main Results:
- The Lysotracker assay successfully measured enlarged lysosomes in patient cells.
- The assay was validated using methyl-β-cyclodextrin in Niemann Pick C disease fibroblasts.
- Demonstrated the assay's utility in identifying compounds that reduce lysosome size.
Conclusions:
- The Lysotracker assay is a viable tool for compound screening in drug development for lysosomal storage diseases.
- This assay facilitates the identification of lead compounds targeting enlarged lysosomes.
- Provides a new method for therapeutic development against lysosomal disorders.
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