Abstract
Impaired efferocytosis has been shown to be associated with, and even to contribute to progression of, chronic inflammatory diseases such as atherosclerosis. Enhancing efferocytosis has been proposed as strategy to treat diseases involving inflammation. Here we present the strategy to increase ‘eat me’ signals on the surface of apoptotic cells by targeting cell surface-expressed phosphatidylserine (PS) with a variant of annexin A5 (Arg-Gly-Asp–annexin A5, RGD–anxA5) that has gained the function to interact with αvβ3 receptors of the phagocyte. We describe design and characterization of RGD–anxA5 and show that introduction of RGD transforms anxA5 from an inhibitor into a stimulator of efferocytosis. RGD–anxA5 enhances engulfment of apoptotic cells by phorbol-12-myristate-13-acetate-stimulated THP-1 (human acute monocytic leukemia cell line) cells in vitro and resident peritoneal mouse macrophages in vivo. In addition, RGD–anxA5 augments secretion of interleukin-10 during efferocytosis in vivo, thereby possibly adding to an anti-inflammatory environment. We conclude that targeting cell surface-expressed PS is an attractive strategy for treatment of inflammatory diseases and that the rationally designed RGD–anxA5 is a promising therapeutic agent.
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Accession codes
Abbreviations
- cRGD:
-
cyclic Arg-Gly-Asp
- EDTA:
-
ethylenediaminetetraacetic acid
- EGTA:
-
ethylene glycol tetraacetic acid
- IL-10:
-
interleukin-10
- MALDI-TOF/TOF:
-
matrix-assisted laser desorption/ionization-time of flight/time of flight
- MFG-E8:
-
milk fat globule-epidermal growth factor 8
- PMA:
-
phorbol-12-myristate-13-acetate
- PS:
-
phosphatidylserine
- RGD–anxA5:
-
Arg-Gly-Asp–annexin A5
- RGT–anxA5:
-
Arg-Gly-Thr–annexin A5
- THP-1:
-
human acute monocytic leukemia cell line
- TIM-4:
-
T-cell immunoglobulin- and mucin-domain-containing molecule-4
- TNFα:
-
tumor necrosis factorα
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Acknowledgements
This study is supported in part by the European Union grant Euregional PACT II from the Interreg IV program of Grensregio Vlaanderen-Nederland (IVA-VLANED-1.20) and by project grants from the Fund for Scientific Research Flanders (FWO-Vlaanderen, G.0728.10, G067512N to DVK) and Methusalem grant (BOF09/01M00709 to PV) from the Flemish Government. DVK is a postdoctoral fellow paid by fellowship from FWO-Vlaanderen. We are indebted to Nicole Bitsch for her technical assistance and to Dr. Gerry Nicolaes (Department Biochemistry, Molecular Modeling and Structure Analysis, Maastricht University) for preparing Figures 1a and b. Lactadherin was obtained as a kind gift from Dr. J Rasmussen.
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Schutters, K., Kusters, D., Chatrou, M. et al. Cell surface-expressed phosphatidylserine as therapeutic target to enhance phagocytosis of apoptotic cells. Cell Death Differ 20, 49–56 (2013). https://doi.org/10.1038/cdd.2012.107
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DOI: https://doi.org/10.1038/cdd.2012.107
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