Design and characterization of a cleavage-resistant Annexin A1 mutant to control inflammation in the microvasculature

Magali Pederzoli-Ribeil; Francesco Maione; Dianne Cooper; Adam Al-Kashi; Jesmond Dalli; Mauro Perretti; Fulvio D'Acquisto

doi:10.1182/blood-2010-02-270520

Design and characterization of a cleavage-resistant Annexin A1 mutant to control inflammation in the microvasculature

Magali Pederzoli-Ribeil, Francesco Maione, Dianne Cooper, Adam Al-Kashi, Jesmond Dalli, Mauro Perretti, Fulvio D'Acquisto

Research output: Contribution to journal › Article › peer-review

Abstract

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.

Original language	English
Pages (from-to)	4288-96
Number of pages	9
Journal	Blood
Volume	116
Issue number	20
DOIs	https://doi.org/10.1182/blood-2010-02-270520
Publication status	Published - 18 Nov 2010

Keywords

Amino Acid Sequence
Animals
Annexin A1
Anti-Inflammatory Agents
Cell Adhesion
Cell Communication
Cell Movement
Endothelial Cells
Female
HEK293 Cells
Humans
Inflammation
Male
Mice
Microvessels
Molecular Sequence Data
Mutant Proteins
Neutrophils
Protein Binding
Protein Engineering
Receptors, Formyl Peptide
Receptors, Lipoxin
Recombinant Proteins
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

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10.1182/blood-2010-02-270520

Cite this

@article{9d3f6589201e49788a33906e53b20ce1,

title = "Design and characterization of a cleavage-resistant Annexin A1 mutant to control inflammation in the microvasculature",

abstract = "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.",

keywords = "Amino Acid Sequence, Animals, Annexin A1, Anti-Inflammatory Agents, Cell Adhesion, Cell Communication, Cell Movement, Endothelial Cells, Female, HEK293 Cells, Humans, Inflammation, Male, Mice, Microvessels, Molecular Sequence Data, Mutant Proteins, Neutrophils, Protein Binding, Protein Engineering, Receptors, Formyl Peptide, Receptors, Lipoxin, Recombinant Proteins, Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't",

author = "Magali Pederzoli-Ribeil and Francesco Maione and Dianne Cooper and Adam Al-Kashi and Jesmond Dalli and Mauro Perretti and Fulvio D'Acquisto",

year = "2010",

month = nov,

day = "18",

doi = "10.1182/blood-2010-02-270520",

language = "English",

volume = "116",

pages = "4288--96",

journal = "Blood",

issn = "0006-4971",

publisher = "American Society of Hematology",

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TY - JOUR

T1 - Design and characterization of a cleavage-resistant Annexin A1 mutant to control inflammation in the microvasculature

AU - Pederzoli-Ribeil, Magali

AU - Maione, Francesco

AU - Cooper, Dianne

AU - Al-Kashi, Adam

AU - Dalli, Jesmond

AU - Perretti, Mauro

AU - D'Acquisto, Fulvio

PY - 2010/11/18

Y1 - 2010/11/18

N2 - 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.

AB - 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.

KW - Amino Acid Sequence

KW - Animals

KW - Annexin A1

KW - Anti-Inflammatory Agents

KW - Cell Adhesion

KW - Cell Communication

KW - Cell Movement

KW - Endothelial Cells

KW - Female

KW - HEK293 Cells

KW - Humans

KW - Inflammation

KW - Male

KW - Mice

KW - Microvessels

KW - Molecular Sequence Data

KW - Mutant Proteins

KW - Neutrophils

KW - Protein Binding

KW - Protein Engineering

KW - Receptors, Formyl Peptide

KW - Receptors, Lipoxin

KW - Recombinant Proteins

KW - Journal Article

KW - Research Support, N.I.H., Extramural

KW - Research Support, Non-U.S. Gov't

U2 - 10.1182/blood-2010-02-270520

DO - 10.1182/blood-2010-02-270520

M3 - Article

C2 - 20705760

SN - 0006-4971

VL - 116

SP - 4288

EP - 4296

JO - Blood

JF - Blood

IS - 20

ER -