title: Monomeric C-reactive protein and Notch-3 co-operatively increase angiogenesis through PI3K signalling pathway.
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(TM) 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.
authors: Healthcare Science Research Institute, Manchester Metropolitan University, Manchester M1 5GD, UK.
authors: Emhamed
authors: E
authors: Boras
authors: Healthcare Science Research Institute, Manchester Metropolitan University, Manchester M1 5GD, UK.
authors: Mark
authors: M
authors: Slevin
authors: Healthcare Science Research Institute, Manchester Metropolitan University, Manchester M1 5GD, UK; Institute of Cardiovascular Sciences, The University of Manchester, Manchester Academic Health Science Centre, Manchester M13 9NT, UK.
authors: M Yvonne
authors: MY
authors: Alexander
authors: Healthcare Science Research Institute, Manchester Metropolitan University, Manchester M1 5GD, UK.
authors: Ali
authors: A
authors: Aljohi
authors: Healthcare Science Research Institute, Manchester Metropolitan University, Manchester M1 5GD, UK.
authors: William
authors: W
authors: Gilmore
authors: Healthcare Science Research Institute, Manchester Metropolitan University, Manchester M1 5GD, UK.
authors: Jason
authors: J
authors: Ashworth
authors: Healthcare Science Research Institute, Manchester Metropolitan University, Manchester M1 5GD, UK; Hospital Universitari Mútua de Terrassa, Department of Neurology, Cerebrovascular Diseases Unit, Terrassa, Barcelona, Spain.
authors: Jerzy
authors: J
authors: Krupinski
authors: Roosevelt University College of Pharmacy, Schaumburg, IL, USA.
authors: Lawrence A
authors: LA
authors: Potempa
authors: Medical Genomics Research Department, King Abdullah International Medical Research Center, Ministry of National Guard Health Affairs, Riyadh 11426, Saudi Arabia.
authors: Ibrahim
authors: I
authors: Al Abdulkareem
authors: Medical Genomics Research Department, King Abdullah International Medical Research Center, Ministry of National Guard Health Affairs, Riyadh 11426, Saudi Arabia.
authors: Adila
authors: A
authors: Elobeid
authors: Healthcare Science Research Institute, Manchester Metropolitan University, Manchester M1 5GD, UK. Electronic address: matouepnasrisa@ngha.med.sa.
authors: Sabine
authors: S
authors: Matou-Nasri
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002097:C-Reactive Protein / Q000494:pharmacology*; D015820:Cadherins / Q000378:metabolism; D002417:Cattle; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002645:Chickens; D002867:Chromones / Q000494:pharmacology; D018920:Coculture Techniques; D004151:Dipeptides / Q000494:pharmacology; D015536:Down-Regulation / Q000187:drug effects; D004587:Electrophoresis, Agar Gel; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D009025:Morpholines / Q000494:pharmacology; D032389:Myocytes, Smooth Muscle / Q000166:cytology; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D051381:Rats; D051880:Receptors, Notch / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects*; D018874:Spheroids, Cellular / Q000166:cytology / Q000187:drug effects; D015854:Up-Regulation / Q000187:drug effects
chemical_list: D015820:Cadherins; D002867:Chromones; D004151:Dipeptides; D009025:Morpholines; C419410:N-(N-(3,5-difluorophenacetyl)alanyl)phenylglycine tert-butyl ester; D051880:Receptors, Notch; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; D002097:C-Reactive Protein; D052250:Nitric Oxide Synthase Type III; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt
keywords: Angiogenesis; C-reactive protein; Gamma-secretase; Notch; PI3K/Akt pathway