Characterizing the anti-inflammatory and tissue protective actions of a novel Annexin A1 peptide

PLOS ONE, Dec 2019

Inflammation in now appreciated to be at the centre of may diseases that affect Western civilization. Current therapeutics for managing these conditions may interfere with the host response leading to immune suppression. We recently developed an annexin (Anx) A1-derived peptide, coined CR-AnxA12-50, which displays potent pro-resolving and tissue protective actions. Herein, we designed a novel peptide using CR-AnxA12-50 as a template that was significantly more resistant to neutrophil-mediated degradation. This peptide, termed CR-AnxA12-48, retained high affinity and specificity to the pro-resolving Lipoxin A4 receptor (ALX) with an IC50 of ~20nM. CR-AnxA12-48 dose dependently (100fM-10nM) promoted the efferocytosis of apoptotic neutrophils, an action that was mediated by the murine orthologue of human ALX. The neutrophil-directed actions were also retained with human primary cells were CR-AnxA12-48 reduced human neutrophil recruitment to activated endothelial cells at concentrations as low as 100 pM. This protective action was mediated by human ALX, since incubation of neutrophils with an anti-ALX antibody reversed this anti-inflammatory actions of CR-AnxA12-48. Administration of this peptide to mice during dermal inflammation led to a significant and dose dependent decrease in neutrophil recruitment. This reduction in neutrophil numbers was more pronounced than that displayed by the parent peptide CR-AnxA12-50. CR-AnxA12-48 was also cardioprotecitve reducing infarct size and systemic chemokine (C-C motif) ligand 5 concentration following ischemia reperfusion injury. These findings identify CR-AnxA12-48 as a new ALX agonist that regulates phagocyte responses and displays tissue-protective actions.

A PDF file should load here. If you do not see its contents the file may be temporarily unavailable at the journal website or you do not have a PDF plug-in installed and enabled in your browser.

Alternatively, you can download the file locally and open with any standalone PDF reader:

https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0175786&type=printable

Characterizing the anti-inflammatory and tissue protective actions of a novel Annexin A1 peptide

April Characterizing the anti-inflammatory and tissue protective actions of a novel Annexin A1 peptide Mauro Perretti 0 1 Clara Di Filippo 1 Michele D'Amico 1 Jesmond Dalli 0 1 0 The William Harvey Research Institute, Barts and The London School of Medicine, Queen Mary University of London , Charterhouse Square, London , United Kingdom , 2 Department of Experimental Medicine, Second University of Naples , Naples , Italy 1 Editor: John Wallace, University of Calgary , CANADA Inflammation in now appreciated to be at the centre of may diseases that affect Western civilization. Current therapeutics for managing these conditions may interfere with the host response leading to immune suppression. We recently developed an annexin (Anx) A1derived peptide, coined CR-AnxA12-50, which displays potent pro-resolving and tissue protective actions. Herein, we designed a novel peptide using CR-AnxA12-50 as a template that was significantly more resistant to neutrophil-mediated degradation. This peptide, termed CR-AnxA12-48, retained high affinity and specificity to the pro-resolving Lipoxin A4 receptor (ALX) with an IC50 of ~20nM. CR-AnxA12-48 dose dependently (100fM-10nM) promoted the efferocytosis of apoptotic neutrophils, an action that was mediated by the murine orthologue of human ALX. The neutrophil-directed actions were also retained with human primary cells were CR-AnxA12-48 reduced human neutrophil recruitment to activated endothelial cells at concentrations as low as 100 pM. This protective action was mediated by human ALX, since incubation of neutrophils with an anti-ALX antibody reversed this anti-inflammatory actions of CR-AnxA12-48. Administration of this peptide to mice during dermal inflammation led to a significant and dose dependent decrease in neutrophil recruitment. This reduction in neutrophil numbers was more pronounced than that displayed by the parent peptide CRAnxA12-50. CR-AnxA12-48 was also cardioprotecitve reducing infarct size and systemic chemokine (C-C motif) ligand 5 concentration following ischemia reperfusion injury. These findings identify CR-AnxA12-48 as a new ALX agonist that regulates phagocyte responses and displays tissue-protective actions. - Data Availability Statement: All relevant data are within the paper. Funding: This work was supported by a collaborative project between Unigene Corp. and Queen Mary University of London and by the William Harvey Research Foundation. JD is supported by a Sir Henry Dale Fellowship jointly funded by the Wellcome Trust and the Royal Society (grant no: 107613/Z/15/Z). MP was supported by the Wellcome Trust (grant no: 086867/Z/08). The funders had no role in study Introduction Inflammation is intrinsically host protective [ 1 ]. Recent evidence suggests that when resolution mechanisms become dysregulated the inflammatory response may be perpetuated leading to unabated inflammation and tissue destruction [ 2, 3 ]. A failed resolution response is now thought to be at least contributory to the onset and propagation of many inflammatory design, data collection and analysis, decision to publish, or preparation of the manuscript. conditions afflicting western civilization including cardiovascular disease [4] and rheumatoid arthritis [ 5 ]. Therapeutics employed in the clinic to date to treat these inflammatory conditions aim to inhibit various mediators that promote the immune response. While this approach is effective at limiting inflammation in some instances it also carries severe side effects including immunosuppression with an elevated risk of infections [ 6, 7 ]. It is now well appreciated that in self-limited inflammation, i.e. when inflammation does not progress to chronicity, the body engages mechanisms that actively downregulate the production of inflammatory mediators as well as the clearance of leukocytes and cellular debris from the site of inflammation [ 8, 9 ]. In this context several families of molecules including specialized pro-resolving mediators [9], gaseous mediators (e.g. carbon monoxide [ 10 ] and hydrogen sulphide [ 11 ]) and proteins were recently described to regulate various aspects of the inflammation-resolution process. Amongst the proteins known to be central to regulating the termination of inflammation is Annexin A1 (AnxA1), a 37 KDa glucocorticoid-regulated protein. This protein regulates leukocyte trafficking in both murine [ 12 ] and human systems [ 13, 14 ], it also promotes the uptake and clearance of apoptotic cells by macrophages [15], a hallmark of resolution, and is organ protective[ 12 ]. The biological actions of this pro-resolving molecule are mediated by the Lipoxin A4 receptor (ALX) [ 8 ]. Mapping of the pharmacophore of this protein to the N-terminal portion lead to the development of peptides that replicate some of the biological actions of the parent protein [ 16 ]. The most widely studied AnxA1-derived peptide is a peptide based on the first 26 amino acid sequence, which d (...truncated)


This is a preview of a remote PDF: https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0175786&type=printable

Mauro Perretti, Clara Di Filippo, Michele D’Amico, Jesmond Dalli. Characterizing the anti-inflammatory and tissue protective actions of a novel Annexin A1 peptide, PLOS ONE, 2017, Volume 12, Issue 4, DOI: 10.1371/journal.pone.0175786