Podocyte Integrin-β3 and Activated Protein C Coordinately Restrict RhoA Signaling and Ameliorate Diabetic Nephropathy.

Abstract

BACKGROUND:

Diabetic nephropathy (dNP), now the leading cause of ESKD, lacks efficient therapies. Coagulation protease-dependent signaling modulates dNP, in part via the G protein-coupled, protease-activated receptors (PARs). Specifically, the cytoprotective protease-activated protein C (aPC) protects from dNP, but the mechanisms are not clear.

METHODS:

A combination of in vitro approaches and mouse models evaluated the role of aPC-integrin interaction and related signaling in dNP.

RESULTS:

The zymogen protein C and aPC bind to podocyte integrin-β3, a subunit of integrin-αvβ3. Deficiency of this integrin impairs thrombin-mediated generation of aPC on podocytes. The interaction of aPC with integrin-αvβ3 induces transient binding of integrin-β3 with G α13 and controls PAR-dependent RhoA signaling in podocytes. Binding of aPC to integrin-β3via its RGD sequence is required for the temporal restriction of RhoA signaling in podocytes. In podocytes lacking integrin-β3, aPC induces sustained RhoA activation, mimicking the effect of thrombin. In vivo, overexpression of wild-type aPC suppresses pathologic renal RhoA activation and protects against dNP. Disrupting the aPC-integrin-β3 interaction by specifically deleting podocyte integrin-β3 or by abolishing aPC's integrin-binding RGD sequence enhances RhoA signaling in mice with high aPC levels and abolishes aPC's nephroprotective effect. Pharmacologic inhibition of PAR1, the pivotal thrombin receptor, restricts RhoA activation and nephroprotects RGE-aPChigh and wild-type mice.Conclusions aPC-integrin-αvβ3 acts as a rheostat, controlling PAR1-dependent RhoA activation in podocytes in diabetic nephropathy. These results identify integrin-αvβ3 as an essential coreceptor for aPC that is required for nephroprotective aPC-PAR signaling in dNP.

• Authors: Thati Madhusudhan, Sanchita Ghosh, Hongjie Wang, Wei Dong, Dheerendra Gupta, Ahmed Elwakiel, Stoyan Stoyanov, Moh'd Mohanad Al-Dabet, Shruthi Krishnan, Ronald Biemann, Sumra Nazir, Silke Zimmermann, Akash Mathew, Ihsan Gadi, Rajiv Rana, Jinyang Zeng-Brouwers, Marcus J Moeller, Liliana Schaefer, Charles T Esmon, Shrey Kohli, Jochen Reiser, Alireza R Rezaie, Wolfram Ruf, Berend Isermann
• Journal: Journal of the American Society of Nephrology : JASN (J Am Soc Nephrol) Vol. 31 Issue 8 Pg. 1762-1780 (08 2020) ISSN: 1533-3450 [Electronic] United States
• PMID: 32709711 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2020 by the American Society of Nephrology.

Chemical References

• Endothelial Protein C Receptor
• GNA13 protein, human
• Integrin beta3
• Protein C
• Receptor, PAR-1
• GTP-Binding Protein alpha Subunits, G12-G13
• rhoA GTP-Binding Protein

Topics

• Animals
• Cytoprotection
• Diabetic Nephropathies (prevention & control)
• Endothelial Protein C Receptor (physiology)
• GTP-Binding Protein alpha Subunits, G12-G13 (physiology)
• HEK293 Cells
• Humans
• Integrin beta3 (physiology)
• Mice
• Mice, Inbred C57BL
• Podocytes (physiology)
• Protein C (physiology)
• Receptor, PAR-1 (physiology)
• rhoA GTP-Binding Protein (physiology)