CD73/Ecto-5 '-nucleotidase protects against vascular inflammation and neointima formation

Institute of Molecular Cardiovascular Research, Rheinisch-Westfälische Technisch Hochschule, Aachen, Germany.
Circulation (Impact Factor: 14.43). 06/2006; 113(17):2120-7. DOI: 10.1161/CIRCULATIONAHA.105.595249
Source: PubMed


Although CD73/ecto-5'-nucleotidase has been implicated in maintaining vasoprotection, its role in regulating endothelial adhesion molecule or inflammatory monocyte recruitment (eg, in the context of vascular injury) remains to be defined.
Compared with wild-type mice, CD73-deficient (CD73(-/-)) mice exhibit increased luminal staining and protein and transcript expression for vascular cell adhesion molecule (VCAM)-1 in carotid arteries. In vitro, aortic endothelial cells (ECs) from CD73(-/-) mice display an upregulation of mRNA and protein expression of VCAM-1, associated with increased nuclear factor (NF)-kappaB activity, as determined by chromatin cross-linking and immunoprecipitation or quantitative p65 binding assays. CD73(-/-) ECs and carotid arteries perfused ex vivo supported increased monocyte arrest under flow conditions, which was mediated by alpha(4beta1) integrin. After wire injury of carotid arteries, CD73 expression and activity were upregulated in wild-type mice, whereas neointimal plaque formation and macrophage content were increased in CD73(-/-) mice versus wild-type mice, concomitant with elevated NF-kappaB activation, luminal VCAM-1 expression, and soluble VCAM-1 concentrations. In contrast, reconstitution of wild-type mice with CD73(-/-) versus CD73(+/+) BM did not significantly exacerbate neointima formation. Treatment with the specific A2A receptor agonist ATL-146e reversed the increased VCAM-1 transcript and protein expression in CD73(-/-) ECs and inhibited monocyte arrest on CD73(-/-) ECs. Continuous infusion of ATL-146e prevented neointima formation in CD73(-/-) mice.
Our data epitomize the importance of vascular CD73 in limiting endothelial activation and monocyte recruitment via generation of adenosine acting through the A2A receptor, providing a molecular basis for therapeutic protection against vascular inflammation and neointimal hyperplasia.

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    • "The role of ecto-5 0 -nucleotidase in endothelial dysfunction and vascular pathologies. Pharmacol Rep (2015), [53]. Additionally to the expression of adhesion molecules, the CCchemokine ligand (also known as MCP-1) produced by vascular cells is pivotal in guiding the recruitment of immune cells. "
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    ABSTRACT: Ecto-5'-nucleotidase (e5NT, CD73) is an enzyme that is highly expressed in endothelium and is involved in the extracellular nucleotide catabolism. CD73 converts AMP to adenosine that via specific subtypes of P1 receptor mediates cytoprotection involving diverse mechanisms such as vasodilatation, suppression of inflammation, inhibition of thrombosis and anti-adrenergic effect. Physiological intravascular concentration of adenosine is in nanomolar range, but could become micromolar in response to various forms of stress. Endothelium is a major site for both CD73 mediated production of adenosine and its cytoprotective effect. Nucleotides (predominantly ATP or ADP) that could be released from different cells via controlled specific of unspecific mechanisms constitute a major source of substrate for adenosine production via CD73. Direct effects of extracellular nucleotides (mediated by P2 receptors) are typically opposite to adenosine P1 mediated activities. Retention of nucleotides and decreased adenosine production due to loss of CD73 function may have negative implications and could be important cause of various pathologies. Protective role of CD73 was indicated in ectopic calcification, atherosclerosis, rejection after xenotransplantation and thrombosis. Reduced activity of CD73 due to lymphocyte contact with endothelium increases its permeability that leads to enhanced leukocyte transmigration. Upregulation of endothelial CD73 may therefore be protective in a number of cardiovascular pathologies. Such effect has been confirmed for some common drugs such as statins and it could be part of its pleiotropic portfolio. Activation of CD73 could be a new target for specific treatment strategy that in particular will enhance endothelial protection. Copyright © 2015. Published by Elsevier Urban & Partner Sp. z o.o.
    Pharmacological reports: PR 05/2015; 67(4). DOI:10.1016/j.pharep.2015.05.002 · 1.93 Impact Factor
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    • "ENTPD1 exerts a vascular protective function against platelet aggregation and vessel occlusion by terminating the prothrombotic and proinflammatory effects of ATP and ADP (Huttinger et al., 2012). E5NT-mediated adenosine production protects against vascular inflammation and neointima formation (Zernecke et al., 2006), diminishes trans-endothelial leukocyte trafficking and mitigates inflammatory and immune sequelae of cardiac transplantation (Hasegawa et al., 2008). "
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    ABSTRACT: We developed an F2A-based multicistronic system to evaluate functional effects of co-expression of three proteins important for xenotransplantation: heme oxygenase 1 (HO1), ecto-5'-nucleotidase (E5NT) and ecto-nucleoside triphosphate diphosphohydrolase-1 (ENTPD1). The tricistronic p2A plasmid that we constructed was able to efficiently drive concurrent expression of HO1, E5NT and ENTPD1 in HEK293T cells. All three overexpressed proteins possessed relevant enzymatic activities, while addition of furin site interfered with protein expression and activity. We conclude that our tricistronic p2A construct is effective and optimal to test the combined protective effects of HO1, E5NT and ENTPD1 against xeno-rejection mechanisms. Copyright © 2015. Published by Elsevier Inc.
    Plasmid 03/2015; 79. DOI:10.1016/j.plasmid.2015.03.004 · 1.58 Impact Factor
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    • "), as well as control of coronary vasodilatation by acting as an endothelium-derived hyperpolarizing factor synthase (Ohta et al., 2013). Studies with ecto-5 0 -nucleotidase/CD73 À/À mice further confirmed the important role of CD73-mediated adenosine formation in microbial infection (Mahamed et al., 2012; Yegutkin et al., 2010), vascular inflammation and neointima formation (Koszalka et al., 2004; Zernecke et al., 2006), arteriogenesis (Boring et al., 2013), atherogenesis (Buchheiser et al., 2011), tumor growth and metastasis (Stagg et al., 2012; Yegutkin et al., 2011b), penile erection (Wen & Xia, 2012), cardioprotection and cardiac allograft vasculopathy (Bonner et al., 2013; Eckle et al., 2007b; Hasegawa et al., 2008), and innate protection during acute lung injury (Eckle et al., 2007a; Kiss et al., 2007). CD73 also controls leukocyte trafficking, particularly mediating its immunomodulatory effects via the modulation of lymphocyte-endothelial adhesion (Koszalka et al., 2004) and subsequent steps of transendothelial leukocyte extravasation (Hasegawa et al., 2008; Henttinen et al., 2003) and migration into draining lymph nodes (Algars et al., 2011; Takedachi et al., 2008). "
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    ABSTRACT: Abstract Extracellular nucleotides and nucleosides mediate diverse signaling effects in virtually all organs and tissues. Most models of purinergic signaling depend on functional interactions between distinct processes, including (i) the release of endogenous ATP and other nucleotides, (ii) triggering of signaling events via a series of nucleotide-selective ligand-gated P2X and metabotropic P2Y receptors as well as adenosine receptors and (iii) ectoenzymatic interconversion of purinergic agonists. The duration and magnitude of purinergic signaling is governed by a network of ectoenzymes, including the enzymes of the nucleoside triphosphate diphosphohydrolase (NTPDase) family, the nucleotide pyrophosphatase/phosphodiesterase (NPP) family, ecto-5'-nucleotidase/CD73, tissue-nonspecific alkaline phosphatase (TNAP), prostatic acid phosphatase (PAP) and other alkaline and acid phosphatases, adenosine deaminase (ADA) and purine nucleoside phosphorylase (PNP). Along with "classical" inactivating ectoenzymes, recent data provide evidence for the co-existence of a counteracting ATP-regenerating pathway comprising the enzymes of the adenylate kinase (AK) and nucleoside diphosphate kinase (NDPK/NME/NM23) families and ATP synthase. This review describes recent advances in this field, with special emphasis on purine-converting ectoenzymes as a complex and integrated network regulating purinergic signaling in such (patho)physiological states as immunomodulation, inflammation, tumorigenesis, arterial calcification and other diseases. The second part of this review provides a comprehensive overview and basic principles of major approaches employed for studying purinergic activities, including spectrophotometric Pi-liberating assays, high-performance liquid chromatographic (HPLC) and thin-layer chromatographic (TLC) analyses of purine substrates and metabolites, capillary electrophoresis, bioluminescent, fluorometric and electrochemical enzyme-coupled assays, histochemical staining, and further emphasizes their advantages, drawbacks and suitability for assaying a particular catalytic reaction.
    Critical Reviews in Biochemistry and Molecular Biology 11/2014; 49(6):473-97. DOI:10.3109/10409238.2014.953627 · 7.71 Impact Factor
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