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Fluorescence-Based Measurements of Phosphatidylserine/Phosphatidylinositol 4-Phosphate Exchange Between Membranes
Published on: March 14, 2021
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Cell-Based Imaging Assay for Detection of Phospholipidosis
Li Zhang1, Shuaizhang Li1, Menghang Xia2
1National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MD, USA.
Methods in Molecular Biology (Clifton, N.J.)
|March 16, 2022
Summary
Drug-induced phospholipidosis, marked by lysosomal phospholipid buildup, can now be screened efficiently. A new high-throughput assay using LipidTOX offers a cost-effective alternative to electron microscopy for identifying causative compounds.
Area of Science:
- Cell biology
- Toxicology
- Drug discovery
Background:
- Drug-induced phospholipidosis is characterized by lysosomal phospholipid accumulation.
- Electron microscopy (EM) is the gold standard for diagnosis but is low-throughput and costly.
- A need exists for efficient screening methods to identify phospholipidosis-inducing compounds.
Purpose of the Study:
- To describe a novel cell-based, high-content assay for detecting phospholipidosis.
- To adapt this assay for high-throughput screening (HTS) applications.
- To validate the assay in relevant cell models.
Main Methods:
- Utilized a cell-based assay with the LipidTOX reagent.
- Implemented a high-content screening platform for miniaturized 1536-well plates.
- Optimized and validated the assay in HepG2 and HepRG cell lines.
Main Results:
- The LipidTOX assay successfully detected cellular phospholipidosis.
- The assay was optimized for HTS, enabling large-scale screening.
- Validation in HepG2 and HepRG cells confirmed assay reliability.
Conclusions:
- A validated, high-throughput, cell-based assay for phospholipidosis has been developed.
- This assay provides a cost-effective and efficient alternative to EM.
- The assay is suitable for screening chemical libraries to identify drug-induced phospholipidosis.
