Annexin A1–containing extracellular vesicles and polymeric nanoparticles promote epithelial wound repair
Giovanna Leoni, … , Andrew S. Neish, Asma Nusrat
Giovanna Leoni, … , Andrew S. Neish, Asma Nusrat
Published February 9, 2015
Citation Information: J Clin Invest. 2015;125(3):1215-1227. https://doi.org/10.1172/JCI76693.
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Research Article Gastroenterology

Annexin A1–containing extracellular vesicles and polymeric nanoparticles promote epithelial wound repair

Abstract

Epithelial restitution is an essential process that is required to repair barrier function at mucosal surfaces following injury. Prolonged breaches in epithelial barrier function result in inflammation and further damage; therefore, a better understanding of the epithelial restitution process has potential for improving the development of therapeutics. In this work, we demonstrate that endogenous annexin A1 (ANXA1) is released as a component of extracellular vesicles (EVs) derived from intestinal epithelial cells, and these ANXA1-containing EVs activate wound repair circuits. Compared with healthy controls, patients with active inflammatory bowel disease had elevated levels of secreted ANXA1-containing EVs in sera, indicating that ANXA1-containing EVs are systemically distributed in response to the inflammatory process and could potentially serve as a biomarker of intestinal mucosal inflammation. Local intestinal delivery of an exogenous ANXA1 mimetic peptide (Ac2-26) encapsulated within targeted polymeric nanoparticles (Ac2-26 Col IV NPs) accelerated healing of murine colonic wounds after biopsy-induced injury. Moreover, one-time systemic administration of Ac2-26 Col IV NPs accelerated recovery following experimentally induced colitis. Together, our results suggest that local delivery of proresolving peptides encapsulated within nanoparticles may represent a potential therapeutic strategy for clinical situations characterized by chronic mucosal injury, such as is seen in patients with IBD.

Authors

Giovanna Leoni, Philipp-Alexander Neumann, Nazila Kamaly, Miguel Quiros, Hikaru Nishio, Hefin R. Jones, Ronen Sumagin, Roland S. Hilgarth, Ashfaqul Alam, Gabrielle Fredman, Ioannis Argyris, Emile Rijcken, Dennis Kusters, Chris Reutelingsperger, Mauro Perretti, Charles A. Parkos, Omid C. Farokhzad, Andrew S. Neish, Asma Nusrat

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Figure 1

Intestinal epithelial wounding induces release of ANXA1-bearing EVs.

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Intestinal epithelial wounding induces release of ANXA1-bearing EVs. (A)...
(A) Immunoblotting for ANXA1 protein in supernatant of nonwounded (nw) or wounded (wound) human epithelial cells. Densitometry analysis shows that ANXA1 protein in cell supernatants derived from wounded cells is significantly increased versus nonwounded confluent cells (2.94-fold increase, P < 0.0001, average of n = 5 immunoblots), and no significant change was observed in the lysate. (B) Immunoblotting for ANXA1 protein in EVs isolated from supernatant. Densitometry analysis shows that ANXA1 protein on EVs derived from wounded cells is increased versus nonwounded cells (2.166-fold increase, P = 0.0066, average of n = 4 immunoblots). (C) Immunogold labeling for ANXA1 and transmission electron microscopy to detect ANXA1 distribution in EVs. Scale bar: 100 nm. (D) Immunoblotting for conventional EV markers in epithelial cell lysates (representative of n = 4 immunoblots). sADAM-10, soluble ADAM-10. (E) EVs were incubated with CD63+ Dynabeads and analyzed by immunoblotting. Densitometry analysis shows that ANXA1 protein on EVs derived from wounded cells is increased versus nonwounded cells (1.459-fold increase, P = 0.0004, average of n = 4 immunoblots). (F) Biotinylated cell surface proteins released from the supernatant of nonwounded and wounded IECs captured with avidin sepharose beads. Densitometry analysis shows that ANXA1 protein on EVs derived from wounded cells is increased versus nonwounded cells (4.239-fold increase increase, P = 0.0002, average of n = 4 immunoblots). (G) Immunoblots show ANXA1 and CD9 proteins in EVs after IEC wounding and treatment with BAPTA-AM (30 μM), Z-VAD-FMK (25 mM), cytochalasin D (2 μM), jasplakinolide (1 μM), dynasore (80 μM), ML-7 (20 μM), and Y-27632 (10 μM). All the results in this figure are representative of at least 3 independent experiments.