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A Multipronged Screening Approach Targeting Inhibition of ETV6 PNT Domain Polymerization
Chloe A N Gerak1, Si Miao Zhang1, Aruna D Balgi1
1Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, BC, Canada.
SLAS Discovery : Advancing Life Sciences R & D
|December 21, 2020
Summary
Researchers developed new experimental and computational assays to find inhibitors of ETV6 PNT domain polymerization, a key process in cancer development. While no inhibitors were identified, the study provides valuable insights for future drug discovery targeting ETV6 oncoproteins.
Area of Science:
- Molecular Biology
- Biochemistry
- Cancer Research
Background:
- ETV6 is a transcriptional repressor whose PNT domain polymerization is crucial for DNA binding.
- Chromosomal translocations involving ETV6 create chimeric oncoproteins driving various cancers.
- No small-molecule inhibitors or assays targeting ETV6 PNT domain polymerization are currently available.
Purpose of the Study:
- To develop novel experimental and computational assays for identifying inhibitors of ETV6 PNT domain polymerization.
- To screen for compounds that disrupt the polymerization process crucial for ETV6-driven oncogenesis.
Main Methods:
- Developed a mammalian cell-based protein-fragment complementation assay using split Gaussia luciferase.
- Established a yeast two-hybrid assay utilizing the HIS3 reporter gene.
- Employed the Bristol University Docking Engine (BUDE) for virtual screening of the ZINC8 library.
- Validated hits using nuclear magnetic resonance (NMR) spectroscopy.
Main Results:
- Successfully established functional mammalian and yeast-based assays for ETV6 PNT domain polymerization.
- Identified over 75 potential ligands through computational screening.
- NMR spectroscopy confirmed that none of the tested compounds directly bind to the purified ETV6 PNT domain.
Conclusions:
- The developed assays provide a foundation for future screening of ETV6 PNT domain polymerization inhibitors.
- This study offers valuable insights that may guide the development of novel therapeutics targeting ETV6 oncoproteins.
- Further research is needed to identify effective inhibitors that disrupt ETV6 PNT domain polymerization for cancer treatment.
