Design and characterization of a cleavage-resistant Annexin A1 mutant to control inflammation in the microvasculature.
Human polymorphonuclear leukocytes adhesion to endothelial cells during the early stage of inflammation leads to cell surface externalization of Annexin A1 (AnxA1), an effector of endogenous anti-inflammation. The antiadhesive properties of AnxA1 become operative to finely tune polymorphonuclear leukocytes transmigration to the site of inflammation. Membrane bound proteinase 3 (PR3) plays a key role in this microenvironment by cleaving the N terminus bioactive domain of AnxA1. In the present study, we generated a PR3-resistant human recombinant AnxA1-named superAnxA1 (SAnxA1)-and tested its in vitro and in vivo properties in comparison to the parental protein. SAnxA1 bound and activated formyl peptide receptor 2 in a similar way as the parental protein, while showing a resistance to cleavage by recombinant PR3. SAnxA1 retained anti-inflammatory activities in the murine inflamed microcirculation (leukocyte adhesion being the readout) and in skin trafficking model. When longer-lasting models of inflammation were applied, SAnxA1 displayed stronger anti-inflammatory effect over time compared with the parental protein. Together these results indicate that AnxA1 cleavage is an important process during neutrophilic inflammation and that controlling the balance between AnxA1/PR3 activities might represent a promising avenue for the discovery of novel therapeutic approaches.
Pederzoli-Ribeil
Magali
M
William Harvey Research Institute, Barts and The London School of Medicine, Queen Mary University of London, London, UK.
Maione
Francesco
F
Cooper
Dianne
D
Al-Kashi
Adam
A
Dalli
Jesmond
J
Perretti
Mauro
M
D'Acquisto
Fulvio
F
D000595:Amino Acid Sequence; D000818:Animals; D017305:Annexin A1 / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D000893:Anti-Inflammatory Agents / Q000378:metabolism; D002448:Cell Adhesion; D002450:Cell Communication; D002465:Cell Movement; D042783:Endothelial Cells / Q000378:metabolism; D005260:Female; D057809:HEK293 Cells; D006801:Humans; D007249:Inflammation / Q000473:pathology* / Q000517:prevention & control*; D008297:Male; D051379:Mice; D055806:Microvessels / Q000378:metabolism / Q000473:pathology*; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000737:chemistry / Q000378:metabolism*; D009504:Neutrophils / Q000166:cytology / Q000378:metabolism; D011485:Protein Binding; D015202:Protein Engineering; D044042:Receptors, Formyl Peptide / Q000378:metabolism; D044067:Receptors, Lipoxin / Q000378:metabolism; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism
D017305:Annexin A1; D000893:Anti-Inflammatory Agents; C105101:FPR2 protein, human; D050505:Mutant Proteins; D044042:Receptors, Formyl Peptide; D044067:Receptors, Lipoxin; D011994:Recombinant Proteins