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Updated: May 28, 2026
Simultaneous Measurement of HDAC1 and HDAC6 Activity in HeLa Cells Using UHPLC-MS
Published on: August 10, 2017
Identification of selective class II histone deacetylase inhibitors using a novel dual-parameter binding assay based
Patricia Haus1, Michael Korbus, Michael Schröder
1Department of Chemical Engineering and Biotechnology, University of Applied Sciences Darmstadt, Darmstadt, Germany.
Abstract:
Histone deacetylases (HDACs) are important epigenetic factors regulating a variety of vital cellular functions such as cell cycle progression, differentiation, cell migration, and apoptosis. Consequently, HDACs have emerged as promising targets for cancer therapy. The drugability of HDACs has been shown by the discovery of several structural classes of inhibitors (HDACis), particularly by the recent approval of two HDACis, vorinostat (ZOLINZA) and romidepsin (Istodax), for the treatment of cutaneous T-cell lymphoma by the US Food and Drug Administration. The outstanding potential of HDACis, with a defined isoform selectivity profile as drugs against a plurality of diseases, vindicates increased effort in developing high-throughput capable assays for screening campaigns. In this study, a dual-competition assay exploiting changes in fluorescence anisotropy and lifetime was used to screen the LOPAC (Sigma-Aldrich, St Louis, MO) library against the bacterial histone deacetylase homologue HDAH from Bordetella, which shares 35% identity with the second deacetylase domain of HDAC6. The binding assay proved to be highly suitable for high-throughput screening campaigns. Several LOPAC compounds have been identified to inhibit HDAH in the lower micromolar range. Most interestingly, some of the hit compounds turned out to be weak but selective inhibitors of human class IIa and IIb HDACs.
Insights
Researchers screened compounds to find new histone deacetylase inhibitors (HDACis). A novel assay identified compounds that inhibit a bacterial HDAC homologue, with some selectively targeting human HDACs for potential cancer therapies.
Area of Science:
- Biochemistry
- Epigenetics
- Drug Discovery
Background:
- Histone deacetylases (HDACs) regulate crucial cellular processes and are key targets in cancer therapy.
- Approved HDAC inhibitors (HDACis) like vorinostat and romidepsin highlight their therapeutic potential.
- Developing high-throughput assays is vital for discovering novel HDACis with specific isoform selectivity.
Purpose of the Study:
- To develop and validate a high-throughput screening assay for identifying histone deacetylase inhibitors (HDACis).
- To screen the LOPAC library against a bacterial histone deacetylase homologue (HDAH) to discover novel inhibitors.
- To identify compounds with potential inhibitory activity against human HDAC isoforms.
Main Methods:
- Utilized a dual-competition assay measuring changes in fluorescence anisotropy and lifetime.
- Screened the LOPAC library against the bacterial HDAH from Bordetella.
- Characterized hit compounds for their inhibitory activity against human class IIa and IIb HDACs.
Main Results:
- The fluorescence-based binding assay demonstrated high suitability for high-throughput screening.
- Identified several LOPAC compounds inhibiting HDAH in the low micromolar range.
- Discovered weak but selective inhibition of human class IIa and IIb HDACs by some identified compounds.
Conclusions:
- A robust high-throughput assay for HDAC inhibitor screening was successfully developed.
- The study identified novel HDAH inhibitors from the LOPAC library.
- Initial findings suggest potential for developing selective inhibitors targeting human HDACs, particularly class IIa and IIb, for therapeutic applications.
