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RAGE-mediated activation of the formin DIAPH1 and human macrophage inflammation are inhibited by a small molecule
Gregory G Theophall1, Michaele B Manigrasso2, Parastou Nazarian1
1Department of Chemistry, State University of New York at Albany, Albany, NY 12222, USA.
Abstract:
RAGE and its intracellular effector molecule, the actin polymerase DIAPH1, mediate inflammation and the complications of diabetes. Using NMR spectroscopy and mass spectrometry, we built a structural model of the RAGE-DIAPH1 complex, revealing how binding of the cytoplasmic tail of RAGE (ctRAGE) to DIAPH1 stimulates its actin polymerization activity, which is inhibited by a small molecule antagonist of RAGE-DIAPH1 interaction, RAGE406R. The solution structure of the RAGE406R - ctRAGE suggests that RAGE406R prevents the formation of the RAGE-DIAPH1. FRET, actin polymerization assays, smooth muscle cell migration, and THP1 cell inflammation experiments, together with the in vivo interrogation of the effects of RAGE406R in mouse models of inflammation and diabetic wound healing, support this mode of RAGE-DIAPH1 antagonism. Finally, the treatment of macrophages differentiated from peripheral blood-derived mononuclear cells from humans with type 1 diabetes with RAGE406R reduces the mRNA expression of the chemokine CCL2, diminishing the expression of a key node in the inflammatory response.
Insights
Researchers identified a small molecule, RAGE406R, that inhibits the RAGE-DIAPH1 interaction, reducing inflammation and improving diabetic wound healing in mouse models.
Area of Science:
- Molecular Biology
- Cell Biology
- Immunology
Background:
- Receptor for Advanced Glycation Endproducts (RAGE) and DIAPH1 drive inflammation and diabetes complications.
- Understanding their interaction is crucial for therapeutic development.
Purpose of the Study:
- To elucidate the structural basis of the RAGE-DIAPH1 complex formation.
- To investigate the inhibitory mechanism of RAGE406R on RAGE-DIAPH1 interaction and its downstream effects.
Main Methods:
- NMR spectroscopy and mass spectrometry to model the RAGE-DIAPH1 complex.
- FRET assays, actin polymerization assays, cell migration, and inflammation experiments.
- In vivo studies in mouse models of inflammation and diabetic wound healing.
Main Results:
- A structural model revealed how RAGE binding to DIAPH1 activates actin polymerization.
- RAGE406R was shown to prevent RAGE-DIAPH1 complex formation.
- RAGE406R demonstrated efficacy in reducing inflammation and improving wound healing in vivo.
- Treatment with RAGE406R decreased CCL2 mRNA expression in human macrophages.
Conclusions:
- RAGE-DIAPH1 interaction is a key target for managing inflammation and diabetes complications.
- RAGE406R represents a promising therapeutic strategy for RAGE-DIAPH1-mediated diseases.
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