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Published on: December 20, 2017
Affinity chromatography reveals direct binding of the GATA4-NKX2-5 interaction inhibitor (3i-1000) with GATA4
Mikael Jumppanen1, Sini M Kinnunen2, Matej Zore1
1Drug Research Program, Division of Pharmaceutical Chemistry and Technology, Faculty of Pharmacy, University of Helsinki, Viikinkaari 5 E, (P.O. Box 56), FI-00014, Helsinki, Finland.
Insights
Researchers identified a compound that protects the heart by directly binding to the GATA4 transcription factor. This finding offers a new approach for treating heart failure by targeting cardiac remodeling.
Area of Science:
- Cardiovascular Research
- Molecular Biology
- Drug Discovery
Background:
- Heart failure is a severe condition with limited treatment options for reversing cardiac remodeling.
- Transcription factors like GATA4 and NKX2-5 play key roles in cardiac hypertrophy and remodeling.
- Existing treatments manage symptoms but do not prevent or reverse heart damage.
Purpose of the Study:
- To investigate the binding site and mechanism of action of a novel cardioprotective compound (1, 3i-1000) targeting GATA4 and NKX2-5.
- To provide direct evidence of compound 1's interaction with its target.
- To explore new therapeutic strategies for heart failure by targeting transcription factors.
Main Methods:
- Synthesis of a small-molecule ligand-binding pulldown probe for compound 1.
- Application of affinity chromatography, Cellular Thermal Shift Assay (CETSA), and AlphaScreen.
- Utilized molecular modeling and intrinsically disordered protein (IDP) knowledge.
Main Results:
- Demonstrated direct physical binding of compound 1 specifically to the GATA4 transcription factor.
- Established compound 1 as a selective GATA4 ligand.
- Overcame challenges associated with targeting disordered transcription factors.
Conclusions:
- Compound 1 selectively binds to GATA4, providing a potential therapeutic strategy for heart failure.
- Advances in technology enable the development of drugs targeting transcription factors, including those involved in cardiac remodeling.
- This study offers a new avenue for preventing and reversing heart damage in heart failure patients.
Abstract:
Heart failure is a serious medical condition with a poor prognosis. Current treatments can only help manage the symptoms and slow the progression of heart failure. However, there is currently no cure to prevent and reverse cardiac remodeling. Transcription factors are in a central role in various cellular processes, and in the heart, GATA4 and NKX2-5 transcription factors mediate hypertrophic responses and remodeling. We have identified compounds that modulate the synergistic interaction of GATA4 and NKX2-5 and shown that the most promising compound (1, 3i-1000) is cardioprotective in vitro and in vivo. However, direct evidence of its binding site and mechanism of action has not been available. Due to the disordered nature of transcription factors, classical target engagement approaches cannot be utilized. Here, we synthesized a small-molecule ligand-binding pulldown probe of compound 1 to utilize affinity chromatography alongside CETSA, AlphaScreen, and molecular modeling to study ligand binding. These results provide the first evidence of direct physical binding of compound 1 selectively to GATA4. While developing drugs that target transcription factors presents challenges, advances in technologies and knowledge of intrinsically disordered proteins enable the identification of small molecules that can selectively target transcription factors.
