title: Cross-talk between macrophages and smooth muscle cells impairs collagen and metalloprotease synthesis and promotes angiogenesis.
abstract: Coronary atherosclerosis complicated by plaque disruption and thrombosis is a critical event in myocardial infarction and stroke, the major causes of cardiovascular death. In atherogenesis, macrophages (MAC) and smooth muscle cells (SMC) are key actors; they synthesize matrix components and numerous factors involved in the process. Here, we design experiments to investigate whether SMC-MAC communication induces changes in ECM protein composition and/or neo-angiogenesis. Cell to cell communication was achieved using trans-well chambers, where SMCs were grown in the upper chamber and differentiated MAC in the bottom chamber for 24 or 72h. We found that cross-talk between MAC and SMC during co-culture: (i) significantly decreased the expression of ECM proteins (collagen I, III, elastin) in SMC; (ii) increased the expression and activity of metalloprotease MMP-9 and expression of collagenase MMP-1, in both MAC and SMC; (iii) augmented the secretion of soluble VEGF in the conditioned media of cell co-culture and VEGF gene expression in both cell types, compared with control cells. Moreover, the conditioned media collected from MAC-SMC co-culture promoted endothelial cell tube formation in Matrigel, signifying an increased angiogenic effect. In addition, the MAC-SMC communication led to an increase in inflammatory IL-1β and TLR-2, which could be responsible for cellular signaling. In conclusion, MAC-SMC communication affects factors and molecules that could alter ECM composition and neo-angiogenesis, features that could directly dictate the progression of atheroma towards the vulnerable plaque. Targeting the MAC-SMC cross-talk may represent a novel therapeutic strategy to slow-down or retard the plaque progression.
authors.affiliation: Institute of Cellular Biology and Pathology "Nicolae Simionescu", Biopathology and Therapy of Inflammation, Bucharest, Romania. Electronic address: elena.dragomir@icbp.ro.
authors.forename: E
authors.initials: E
authors.lastname: Butoi
authors.affiliation: Institute of Cellular Biology and Pathology "Nicolae Simionescu", Biopathology and Therapy of Inflammation, Bucharest, Romania; Laboratory of Molecular Medical Biochemistry, Nencki Institute of Experimental Bilogy, Warsaw, Poland. Electronic address: anca.gan@icbp.ro.
authors.forename: A M
authors.initials: AM
authors.lastname: Gan
authors.affiliation: Institute of Cellular Biology and Pathology "Nicolae Simionescu", Biopathology and Therapy of Inflammation, Bucharest, Romania. Electronic address: monica.pirvulescu@icbp.ro.
authors.forename: M M
authors.initials: MM
authors.lastname: Tucureanu
authors.affiliation: Institute of Cellular Biology and Pathology "Nicolae Simionescu", Biopathology and Therapy of Inflammation, Bucharest, Romania. Electronic address: daniela.stan@icbp.ro.
authors.forename: D
authors.initials: D
authors.lastname: Stan
authors.affiliation: Institute of Cellular Biology and Pathology "Nicolae Simionescu", Biopathology and Therapy of Inflammation, Bucharest, Romania. Electronic address: macarierazvan@gmail.com.
authors.forename: R D
authors.initials: RD
authors.lastname: Macarie
authors.affiliation: Institute of Cellular Biology and Pathology "Nicolae Simionescu", Biopathology and Therapy of Inflammation, Bucharest, Romania. Electronic address: cristina.constantinescu@icbp.ro.
authors.forename: C
authors.initials: C
authors.lastname: Constantinescu
authors.affiliation: Institute of Cellular Biology and Pathology "Nicolae Simionescu", Biopathology and Therapy of Inflammation, Bucharest, Romania. Electronic address: manuela.calin@icbp.ro.
authors.forename: M
authors.initials: M
authors.lastname: Calin
authors.affiliation: Institute of Cellular Biology and Pathology "Nicolae Simionescu", Biopathology and Therapy of Inflammation, Bucharest, Romania. Electronic address: maya.simionescu@icbp.ro.
authors.forename: M
authors.initials: M
authors.lastname: Simionescu
authors.affiliation: Institute of Cellular Biology and Pathology "Nicolae Simionescu", Biopathology and Therapy of Inflammation, Bucharest, Romania. Electronic address: ileana.manduteanu@icbp.ro.
authors.forename: I
authors.initials: I
authors.lastname: Manduteanu
mesh_terms: D050197:Atherosclerosis / Q000201:enzymology* / Q000235:genetics / Q000473:pathology / Q000503:physiopathology; D002450:Cell Communication; D002460:Cell Line; D018920:Coculture Techniques; D003094:Collagen / Q000378:metabolism*; D017077:Culture Media, Conditioned / Q000378:metabolism; D006801:Humans; D018836:Inflammation Mediators / Q000378:metabolism; D053583:Interleukin-1beta / Q000378:metabolism; D008264:Macrophages / Q000201:enzymology* / Q000473:pathology; D020781:Matrix Metalloproteinase 1 / Q000378:metabolism*; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism*; D009131:Muscle, Smooth, Vascular / Q000201:enzymology* / Q000473:pathology / Q000503:physiopathology; D032389:Myocytes, Smooth Muscle / Q000201:enzymology* / Q000473:pathology; D018919:Neovascularization, Physiologic; D034622:RNA Interference; D015398:Signal Transduction; D013997:Time Factors; D051195:Toll-Like Receptor 2 / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
chemical_list: D017077:Culture Media, Conditioned; C508585:IL1B protein, human; D018836:Inflammation Mediators; D053583:Interleukin-1beta; C495342:TLR2 protein, human; D051195:Toll-Like Receptor 2; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D003094:Collagen; C579270:MMP9 protein, human; D020780:Matrix Metalloproteinase 9; C522360:MMP1 protein, human; D020781:Matrix Metalloproteinase 1
keywords: Angiogenesis; Atherosclerosis; Extracellular matrix proteins; Macrophage–SMC cross-talk; Matrix metalloproteinases