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Updated: Mar 21, 2026
Simultaneous Measurement of HDAC1 and HDAC6 Activity in HeLa Cells Using UHPLC-MS
Published on: August 10, 2017
Cell-based multi-substrate assay coupled to UHPLC-ESI-MS/MS for a quick identification of class-specific HDAC
Vincent Zwick1, Claudia Simões-Pires1, Muriel Cuendet1
1a School of Pharmaceutical Sciences, University of Geneva, University of Lausanne , Geneva , Switzerland.
Abstract:
Histone deacetylases (HDAC) are involved in several diseases including cancer, cardiovascular and neurodegenerative disorders, and the search for inhibitors is a current topic in drug discovery. Four HDAC inhibitors have already been approved by the FDA for cancer therapy and others are under clinical studies. However, the clinical utility of some of them is limited because of unfavorable toxicities associated with their broad range of HDAC inhibitory effects. Toxicity could be decreased by using HDAC inhibitors with improved specificity. To date, the most popular screening assays are based on fluorescence-labeled substrates incubated with an enzymatic source (cells extracts or recombinant isoforms). Here, we describe a high-throughput cell-based UHPLC-ESI-MS/MS assay able to rapidly predict activity against HDAC1 and HDAC6 in a cell environment. This method is predicted to be a useful tool to accelerate the search for class-selective HDAC inhibitors in drug discovery.
Insights
This study introduces a new cell-based assay for quickly screening histone deacetylase (HDAC) inhibitors. The method aids in discovering more specific HDAC inhibitors to reduce drug toxicity in cancer and other diseases.
Area of Science:
- Biochemistry
- Pharmacology
- Drug Discovery
Background:
- Histone deacetylases (HDACs) are implicated in various diseases, including cancer, cardiovascular, and neurodegenerative disorders.
- Current HDAC inhibitors face limitations due to broad inhibitory effects and associated toxicities.
- Developing specific HDAC inhibitors is crucial for improving therapeutic efficacy and safety.
Purpose of the Study:
- To develop a high-throughput, cell-based assay for predicting HDAC inhibitor activity.
- To enable rapid screening of compounds targeting specific HDAC isoforms (HDAC1 and HDAC6) within a cellular context.
- To accelerate the discovery of class-selective HDAC inhibitors for drug development.
Main Methods:
- Development of a novel ultra-high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UHPLC-ESI-MS/MS) assay.
- Utilizing a cell-based system to assess inhibitor activity directly within a cellular environment.
- High-throughput screening approach for rapid compound evaluation.
Main Results:
- The developed UHPLC-ESI-MS/MS assay demonstrated high throughput for predicting activity against HDAC1 and HDAC6.
- The cell-based nature of the assay provides a more relevant biological context compared to traditional methods.
- The assay successfully predicted activity against target HDAC isoforms in a cellular environment.
Conclusions:
- The novel cell-based UHPLC-ESI-MS/MS assay is a valuable tool for accelerating drug discovery.
- This method facilitates the identification of specific HDAC inhibitors, potentially reducing off-target toxicities.
- The assay supports the development of more effective and safer therapeutics for HDAC-related diseases.
