An orthosteric/allosteric bivalent peptide agonist comprising covalently linked protease-activated receptor-derived peptides mimics in vitro and in vivo activities of activated protein C.

Autor:	Healy LD; Department of Molecular Medicine, The Scripps Research Institute, La Jolla, California, USA., Fernández JA; Department of Molecular Medicine, The Scripps Research Institute, La Jolla, California, USA., Aiolfi R; Department of Molecular Medicine, The Scripps Research Institute, La Jolla, California, USA., Mosnier LO; Department of Molecular Medicine, The Scripps Research Institute, La Jolla, California, USA., Griffin JH; Department of Molecular Medicine, The Scripps Research Institute, La Jolla, California, USA. Electronic address: jgriffin@scripps.edu.
Jazyk:	angličtina
Zdroj:	Journal of thrombosis and haemostasis : JTH [J Thromb Haemost] 2024 Jul; Vol. 22 (7), pp. 2039-2051. Date of Electronic Publication: 2024 Apr 24.
DOI:	10.1016/j.jtha.2024.04.007
Abstrakt:	Background: Activated protein C (APC) has anticoagulant and cytoprotective cell-signaling activities, which often require protease-activated receptor (PAR) 1 and PAR3 and PAR cleavages at noncanonical sites (R46-N47 and R41-G42, respectively). Some PAR1-derived (P1) peptides and PAR3-derived (P3) peptides, eg, P1-47-66 and P3-42-65, mimic APC's cell signaling. In anti-inflammatory assays, these 2 peptides at low concentrations synergistically attenuate cellular inflammation. Objectives: To determine whether a P1 peptide covalently linked to a P3 peptide mimics APC's anti-inflammatory and endothelial barrier stabilization activities. Methods: Anti-inflammatory assays employed stimulated THP-1 cells and caspase-1 measurements. Cultured human EA.hy926 or murine aortic endothelial cells (ECs) exposed to thrombin were monitored for transendothelial electrical resistance. Bivalent covalently linked P1:P3 peptides were studied for APC-like activities. Results: In anti-inflammatory assays, P1-47-55 was as active as P1-47-66 and some P3 peptides (eg, P3-44-54 and P3-51-65) were as active as P3-42-65. The bivalent P1:P3 peptide comprising P1-47-55-(Gly[10 residues])-P3-51-65 (designated "G10 peptide") was more potently anti-inflammatory than the P1 or P3 peptide alone. In transendothelial electrical resistance studies of thrombin-challenged ECs, P1-47-55 and the G10 peptide mimicked APC's protective actions. In dose-response studies, the G10 peptide was more potent than the P1-47-55 peptide. In murine EC studies, the murine PAR-sequence-derived G10 peptide mimicked murine APC's activity. Anti-PAR1 and anti-PAR3 antibodies, but not anti-endothelial protein C receptor antibodies, abated G10's cytoprotection, showing that G10's actions involve PAR1:PAR3. G10 significantly increased survival in murine endotoxemia. Conclusion: The PAR-sequence-derived G10 peptide is a bivalent agonist that mimics APC's cytoprotective, anti-inflammatory, and endothelial barrier-stabilizing actions and APC's protection against endotoxemic mortality. Competing Interests: Declaration of competing interests L.D.H., L.O.M., and J.H.G. are coinventors of Scripps-owned patents related to some studies in this report. L.O.M. and J.H.G. are on the Scientific Advisory Board of Novapep Pty Ltd. R.A. and J.A.F. have no competing interests. (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)
Databáze:	MEDLINE
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