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High-throughput screening assay for PARP-HPF1 interaction inhibitors to affect DNA damage repair

Saurabh S Dhakar1, Albert Galera-Prat1, Lari Lehtiö2

  • 1Faculty of Biochemistry and Molecular Medicine and Biocenter Oulu, University of Oulu, Oulu, Finland.

Scientific Reports
|February 16, 2024
PubMed

Insights

Researchers developed a new assay to find drugs targeting the PARP1/2-HPF1 interaction, crucial for DNA repair. This method identified two compounds, Dimethylacrylshikonin and Alkannin, as potential anticancer agents.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Drug Discovery

Background:

  • Poly(ADP-ribose) polymerases (PARP1 and PARP2) are key enzymes in DNA repair, crucial for detecting DNA damage and initiating repair pathways.
  • PARP inhibitors are established anticancer drugs, particularly effective in tumors with specific DNA repair deficiencies.
  • Histone PARylation Factor (HPF1) forms a complex with PARP1/2, altering substrate specificity and playing a critical role in DNA damage response.

Purpose of the Study:

  • To develop a high-throughput screening (HTS) assay to identify inhibitors of the PARP1/2-HPF1 interaction.
  • To validate the assay's robustness and utility for drug discovery targeting the PARP-HPF1 complex.

Main Methods:

  • Development and optimization of a Förster Resonance Energy Transfer (FRET)-based HTS assay to monitor PARP1/2-HPF1 interaction.
  • Verification of the FRET signal and interaction by competitive disruption.
  • Screening of inhibitor libraries against the PARP-HPF1 complex.

Main Results:

  • The FRET-based assay was optimized for signal detection and validated for robustness and automation.
  • Two compounds, Dimethylacrylshikonin and Alkannin, were identified as inhibitors of the PARP1/2-HPF1 interaction with micromolar potency.
  • The assay demonstrated reliable performance in identifying inhibitors of the target complex.

Conclusions:

  • The developed FRET assay is a robust and automatable tool for discovering inhibitors of the PARP1/2-HPF1 interaction.
  • This approach offers a novel strategy for developing new anticancer agents by targeting the PARP-HPF1 complex.
  • Identified compounds provide a starting point for further drug development against cancer

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