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Updated: Aug 14, 2026
Enrichment of Mammalian Tissues and Xenopus Oocytes with Cholesterol
Published on: March 25, 2020
Automated microscopy screening for compounds that partially revert cholesterol accumulation in Niemann-Pick C cells
Nina H Pipalia1, Amy Huang, Harold Ralph
1Department of Biochemistry, Weill Medical College of Cornell University, New York, NY 10021, USA.
Abstract:
Niemann-Pick disease type C (NPC) is an autosomal recessive genetic disorder manifested by abnormal accumulation of unesterified cholesterol and other lipids. We screened combinatorially synthesized chemical libraries to identify compounds that would partially revert cholesterol accumulation. Cultured CHO cells with NPC phenotypes (CT60 and CT43) were used for screening along with normal CHO cells as a control. We developed an automated microscopy assay based on imaging of filipin fluorescence for estimating cholesterol accumulation in lysosomal storage organelles. Our primary screen of 14,956 compounds identified 14 hit compounds that caused significant reduction in cellular cholesterol accumulation at 10 microM. We then screened a secondary library of 3,962 compounds selected based on chemical similarity to the initial hits and identified 7 compounds that demonstrated greater efficacy and lower toxicity than the original hits. These compounds are effective at concentrations of 123 nM to 3 microM in reducing the cholesterol accumulation in cells with a NPC1 phenotype.
Insights
Researchers screened chemical libraries to find compounds that reduce cholesterol buildup in Niemann-Pick disease type C (NPC). They identified several promising compounds effective at low concentrations, offering potential therapeutic avenues for this genetic disorder.
Area of Science:
- Biochemistry
- Genetics
- Pharmacology
Background:
- Niemann-Pick disease type C (NPC) is a rare, autosomal recessive genetic disorder.
- NPC is characterized by the pathological accumulation of unesterified cholesterol and other lipids within cells.
- This lipid accumulation primarily affects lysosomal storage organelles, leading to cellular dysfunction.
Purpose of the Study:
- To identify novel small molecules capable of partially reversing cholesterol accumulation in NPC.
- To screen large chemical libraries for compounds with therapeutic potential against NPC phenotypes.
- To develop and utilize an automated microscopy assay for efficient screening of cholesterol-reducing compounds.
Main Methods:
- Combinatorial synthesis and screening of chemical libraries against NPC cell models (CT60 and CT43 CHO cells).
- Development of an automated microscopy assay utilizing filipin fluorescence to quantify intracellular cholesterol levels.
- Primary screening of 14,956 compounds followed by secondary screening of 3,962 related compounds based on initial hit efficacy and toxicity.
Main Results:
- The primary screen identified 14 hit compounds that significantly reduced cellular cholesterol accumulation at 10 microM.
- Secondary screening identified 7 compounds with enhanced efficacy and reduced toxicity compared to initial hits.
- These 7 compounds demonstrated effectiveness in reducing cholesterol accumulation in NPC1 phenotype cells at concentrations ranging from 123 nM to 3 microM.
Conclusions:
- The study successfully identified potent small molecules that can partially revert cholesterol accumulation in Niemann-Pick disease type C models.
- These identified compounds represent promising therapeutic candidates for further investigation in NPC treatment.
- The automated microscopy assay proved effective for high-throughput screening of compounds targeting lipid metabolism disorders.
