title: Tropoelastin modulates TGF-β1-induced expression of VEGF and CTGF in airway smooth muscle cells.
abstract: Elastin is predominantly comprised of crosslinked tropoelastin. For many years elastin was considered to serve a solely structural role but is now being increasingly identified as causal in cell signaling, development and repair. We introduced tropoelastin into an in vitro model in which airway smooth muscle cells (ASMCs) were stimulated with transforming growth factor (TGF)-β1 to examine the modulatory effect of this modular elastin sequence on release of angiogenic factors and matrix metalloproteinases (MMPs). Human ASMCs were presented to surfaces coated with tropoelastin or collagen and controls, then stimulated with TGF-β1. Transcript levels of vascular endothelial growth factor (VEGF) and connective tissue growth factor (CTGF) were quantified 4 and 24 h after TGF-β1 stimulation. Protein VEGF release from cells and CTGF sequestered at cell surfaces were measured by ELISA at 24 and 48 h. TGF-β1 increased VEGF mRNA 2.4 fold at 4 h and 5 fold at 24 h, accompanied by elevated cognate protein release 3 fold at 24 h and 2.5 fold at 48 h. TGF-β1 stimulation increased CTGF mRNA 6.9 fold at 4 h and 11.8 fold at 24 h, accompanied by increased sequestering of its protein counterpart 1.2 fold at 24 h and 1.4 fold at 48 h. Pre-incubation of cells with tropoelastin did not modulate VEGF or CTGF mRNA expression, but combined with TGF-β1 stimulation it led to enhanced VEGF release 5.1-fold at 24h and 4.4-fold at 48 h. Pre-incubation with tropoelastin decreased CTGF sequestering 0.6-fold at 24 and 48 h, and increased MMP-2 production. Collagen pre-incubation under the same conditions displayed no effect on TGF-β1 stimulation apart from a slightly decreased (0.9 fold) sequestered CTGF at 48 h. As CTGF is known to anchor VEGF to the matrix and inhibit its angiogenic activity, a process which can be reversed by digestion with MMP-2, these findings reveal that elastin sequences can disrupt the balance of angiogenic factors, with implications for aberrant angiogenesis. The results suggest a model of molecular crosstalk and support an active role for elastin in vascular remodeling.
authors.affiliation: School of Molecular Bioscience, University of Sydney, NSW 2006, Australia. Electronic address: creddel@anzac.edu.au.
authors.forename: Caroline J
authors.initials: CJ
authors.lastname: Reddel
authors.forename: Daniele
authors.initials: D
authors.lastname: Cultrone
authors.forename: Jelena
authors.initials: J
authors.lastname: Rnjak-Kovacina
authors.forename: Anthony S
authors.initials: AS
authors.lastname: Weiss
authors.forename: Janette K
authors.initials: JK
authors.lastname: Burgess
mesh_terms: D003094:Collagen / Q000378:metabolism; D055513:Connective Tissue Growth Factor / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D005786:Gene Expression Regulation / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D032389:Myocytes, Smooth Muscle / Q000378:metabolism / Q000502:physiology*; D012137:Respiratory System / Q000166:cytology; D053773:Transforming Growth Factor beta1 / Q000378:metabolism* / Q000494:pharmacology; D014333:Tropoelastin / Q000502:physiology*; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*
chemical_list: C526317:CCN2 protein, human; D053773:Transforming Growth Factor beta1; D014333:Tropoelastin; D042461:Vascular Endothelial Growth Factor A; D055513:Connective Tissue Growth Factor; D003094:Collagen; D020778:Matrix Metalloproteinase 2
keywords: ASMC; CTGF; Connective tissue growth factor; Elastin; Electrospinning; MMP; TGF; Transforming growth factor β1; Tropoelastin; VEGF; Vascular endothelial growth factor A; airway smooth muscle cell; connective tissue growth factor; matrix metalloproteinase; transforming growth factor; vascular endothelial growth factor