Monomeric C-reactive protein and Notch-3 co-operatively increase angiogenesis through PI3K signalling pathway

Boras, E, Slevin, M ORCID: https://orcid.org/0000-0003-3767-4861, Alexander, MY, Aljohi, A, Gilmore, W, Ashworth, J ORCID: https://orcid.org/0000-0001-7045-7899, Krupinski, J ORCID: https://orcid.org/0000-0002-5136-8898, Potempa, LA, Al Abdulkareem, I, Elobeid, A and Matou-Nasri, S (2014) Monomeric C-reactive protein and Notch-3 co-operatively increase angiogenesis through PI3K signalling pathway. Cytokine, 69 (2). pp. 165-179. ISSN 1096-0023

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Official URL: https://doi.org/10.1016/j.cyto.2014.05.027

Abstract

C-reactive protein (CRP) is the most acute-phase reactant serum protein of inflammation and a strong predictor of cardiovascular disease. Its expression is associated with atherosclerotic plaque instability and the formation of immature micro-vessels. We have previously shown that CRP upregulates endothelial-derived Notch-3, a key receptor involved in vascular development, remodelling and maturation. In this study, we investigated the links between the bioactive monomeric CRP (mCRP) and Notch-3 signalling in angiogenesis. We used in vitro (cell counting, wound-healing and tubulogenesis assays) and in vivo (chorioallantoic membrane) angiogenic assays and Western blotting to study the angiogenic signalling pathways induced by mCRP and Notch-3 activator chimera protein (Notch-3/Fc). Our results showed an additive effect on angiogenesis of mCRP stimulatory effect combined with Notch-3/Fc promoting bovine aortic endothelial cell (BAEC) proliferation, migration, tube formation in Matrigel<sup>TM</sup> with up-regulation of phospho-Akt expression. The pharmacological blockade of PI3K/Akt survival pathway by LY294002 fully inhibited in vitro and in vivo angiogenesis induced by mCRP/Notch-3/Fc combination while blocking Notch signalling by gamma-secretase inhibitor (DAPT) partially inhibited mCRP/Notch-3/Fc-induced angiogenesis. Using a BAEC vascular smooth muscle cell co-culture sprouting angiogenesis assay and transmission electron microscopy, we showed that activation of both mCRP and Notch-3 signalling induced the formation of thicker sprouts which were shown later by Western blotting to be associated with an up-regulation of N-cadherin expression and a down-regulation of VE-cadherin expression. Thus, mCRP combined with Notch-3 activator promote angiogenesis through the PI3K/Akt pathway and their therapeutic combination has potential to promote and stabilize vessel formation whilst reducing the risk of haemorrhage from unstable plaques. © 2014 Elsevier Ltd.

Item Type:	Article
Peer-reviewed:	Yes
Date Deposited:	01 Mar 2017 09:07
Publisher:	Elsevier
Additional Information:	This is an Author Accepted Manuscript of a paper accepted for publication in Cytokine, published by and copyright Elsevier on behalf of the International Cytokine Society
Divisions:	Faculties > Science and Engineering
Subject terms:	Science & Technology, Life Sciences & Biomedicine, Biochemistry & Molecular Biology, Cell Biology, Immunology, C-reactive protein, Notch, Angiogenesis, PI3K/Akt pathway, Gamma-secretase, ENDOTHELIAL-CELLS, SMOOTH-MUSCLE, EXPRESSION, CRP, RECEPTORS, OLIGOSACCHARIDES, INFLAMMATION, ACTIVATION, HEMORRHAGE, LIGANDS, C-reactive protein, Notch, Angiogenesis, PI3K/Akt pathway, Gamma-secretase, Spheroids, Cellular, Endothelial Cells, Myocytes, Smooth Muscle, Animals, Chickens, Cattle, Rats, Morpholines, Chromones, Dipeptides, C-Reactive Protein, Cadherins, Blotting, Western, Electrophoresis, Agar Gel, Coculture Techniques, Signal Transduction, Cell Proliferation, Cell Movement, Down-Regulation, Up-Regulation, Neovascularization, Physiologic, Proto-Oncogene Proteins c-akt, Receptors, Notch, Nitric Oxide Synthase Type III, Phosphatidylinositol 3-Kinases, Angiogenesis, C-reactive protein, Gamma-secretase, Notch, PI3K/Akt pathway, Animals, Blotting, Western, C-Reactive Protein, Cadherins, Cattle, Cell Movement, Cell Proliferation, Chickens, Chromones, Coculture Techniques, Dipeptides, Down-Regulation, Electrophoresis, Agar Gel, Endothelial Cells, Morpholines, Myocytes, Smooth Muscle, Neovascularization, Physiologic, Nitric Oxide Synthase Type III, Phosphatidylinositol 3-Kinases, Proto-Oncogene Proteins c-akt, Rats, Receptors, Notch, Signal Transduction, Spheroids, Cellular, Up-Regulation, 0601 Biochemistry and Cell Biology, 0604 Genetics, 1107 Immunology, Immunology
URI:	https://e-space.mmu.ac.uk/id/eprint/206
DOI:	https://doi.org/10.1016/j.cyto.2014.05.027
ISSN	1096-0023
e-ISSN	1096-0023

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