title: The focal adhesion: a regulated component of aortic stiffness.
abstract: Increased aortic stiffness is an acknowledged predictor and cause of cardiovascular disease. The sources and mechanisms of vascular stiffness are not well understood, although the extracellular matrix (ECM) has been assumed to be a major component. We tested here the hypothesis that the focal adhesions (FAs) connecting the cortical cytoskeleton of vascular smooth muscle cells (VSMCs) to the matrix in the aortic wall are a component of aortic stiffness and that this component is dynamically regulated. First, we examined a model system in which magnetic tweezers could be used to monitor cellular cortical stiffness, serum-starved A7r5 aortic smooth muscle cells. Lysophosphatidic acid (LPA), an activator of myosin that increases cell contractility, increased cortical stiffness. A small molecule inhibitor of Src-dependent FA recycling, PP2, was found to significantly inhibit LPA-induced increases in cortical stiffness, as well as tension-induced increases in FA size. To directly test the applicability of these results to force and stiffness development at the level of vascular tissue, we monitored mouse aorta ring stiffness with small sinusoidal length oscillations during agonist-induced contraction. The alpha-agonist phenylephrine, which also increases myosin activation and contractility, increased tissue stress and stiffness in a PP2- and FAK inhibitor 14-attenuated manner. Subsequent phosphotyrosine screening and follow-up with phosphosite-specific antibodies confirmed that the effects of PP2 and FAK inhibitor 14 in vascular tissue involve FA proteins, including FAK, CAS, and paxillin. Thus, in the present study we identify, for the first time, the FA of the VSMC, in particular the FAK-Src signaling complex, as a significant subcellular regulator of aortic stiffness and stress.
authors.affiliation: Department of Biomedical Engineering, Boston University, Boston, Massachusetts, United States of America.
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Saphirstein
authors.forename: Yuan Z
authors.initials: YZ
authors.lastname: Gao
authors.forename: Mikkel H
authors.initials: MH
authors.lastname: Jensen
authors.forename: Cynthia M
authors.initials: CM
authors.lastname: Gallant
authors.forename: Susanne
authors.initials: S
authors.lastname: Vetterkind
authors.forename: Jeffrey R
authors.initials: JR
authors.lastname: Moore
authors.forename: Kathleen G
authors.initials: KG
authors.lastname: Morgan
mesh_terms: D000818:Animals; D001011:Aorta / Q000187:drug effects / Q000378:metabolism*; D005109:Extracellular Matrix / Q000378:metabolism*; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D022001:Focal Adhesions / Q000187:drug effects; D066298:In Vitro Techniques; D008246:Lysophospholipids / Q000494:pharmacology; D009119:Muscle Contraction; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000378:metabolism*; D009219:Myosin-Light-Chain Kinase / Q000037:antagonists & inhibitors / Q000378:metabolism; D010766:Phosphorylation; D011743:Pyrimidines / Q000494:pharmacology; D051381:Rats; D059289:Vascular Stiffness / Q000187:drug effects; D019061:src-Family Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism
chemical_list: C412373:AG 1879; D008246:Lysophospholipids; D011743:Pyrimidines; D051416:Focal Adhesion Protein-Tyrosine Kinases; D019061:src-Family Kinases; D009219:Myosin-Light-Chain Kinase; C032881:lysophosphatidic acid