title: β3 integrin in cardiac fibroblast is critical for extracellular matrix accumulation during pressure overload hypertrophy in mouse.
abstract: The adhesion receptor β3 integrin regulates diverse cellular functions in various tissues. As β3 integrin has been implicated in extracellular matrix (ECM) remodeling, we sought to explore the role of β3 integrin in cardiac fibrosis by using wild type (WT) and β3 integrin null (β3-/-) mice for in vivo pressure overload (PO) and in vitro primary cardiac fibroblast phenotypic studies. Compared to WT mice, β3-/- mice upon pressure overload hypertrophy for 4 wk by transverse aortic constriction (TAC) showed a substantially reduced accumulation of interstitial fibronectin and collagen. Moreover, pressure overloaded LV from β3-/- mice exhibited reduced levels of both fibroblast proliferation and fibroblast-specific protein-1 (FSP1) expression in early time points of PO. To test if the observed impairment of ECM accumulation in β3-/- mice was due to compromised cardiac fibroblast function, we analyzed primary cardiac fibroblasts from WT and β3-/- mice for adhesion to ECM proteins, cell spreading, proliferation, and migration in response to platelet derived growth factor-BB (PDGF, a growth factor known to promote fibrosis) stimulation. Our results showed that β3-/- cardiac fibroblasts exhibited a significant reduction in cell-matrix adhesion, cell spreading, proliferation and migration. In addition, the activation of PDGF receptor associated tyrosine kinase and non-receptor tyrosine kinase Pyk2, upon PDGF stimulation were impaired in β3-/- cells. Adenoviral expression of a dominant negative form of Pyk2 (Y402F) resulted in reduced accumulation of fibronectin. These results indicate that β3 integrin-mediated Pyk2 signaling in cardiac fibroblasts plays a critical role in PO-induced cardiac fibrosis.
authors.affiliation: Cardiology Division, Department of Medicine, Gazes Cardiac Research Institute, Medical University of South Carolina, Charleston, South Carolina, United States of America.
authors.forename: Sundaravadivel
authors.initials: S
authors.lastname: Balasubramanian
authors.forename: Lakeya
authors.initials: L
authors.lastname: Quinones
authors.forename: Harinath
authors.initials: H
authors.lastname: Kasiganesan
authors.forename: Yuhua
authors.initials: Y
authors.lastname: Zhang
authors.forename: Dorea L
authors.initials: DL
authors.lastname: Pleasant
authors.forename: Kamala P
authors.initials: KP
authors.lastname: Sundararaj
authors.forename: Michael R
authors.initials: MR
authors.lastname: Zile
authors.forename: Amy D
authors.initials: AD
authors.lastname: Bradshaw
authors.forename: Dhandapani
authors.initials: D
authors.lastname: Kuppuswamy
mesh_terms: D000818:Animals; D001011:Aorta / Q000503:physiopathology / Q000601:surgery; D000077214:Becaplermin; D015153:Blotting, Western; D006332:Cardiomegaly / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D002448:Cell Adhesion / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism; D003251:Constriction, Pathologic / Q000503:physiopathology; D005109:Extracellular Matrix / Q000378:metabolism*; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D005353:Fibronectins / Q000378:metabolism; D051418:Focal Adhesion Kinase 2 / Q000378:metabolism; D006984:Hypertrophy; D007150:Immunohistochemistry; D039661:Integrin beta3 / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009206:Myocardium / Q000378:metabolism* / Q000473:pathology; D011312:Pressure; D020574:Proto-Oncogene Proteins c-sis / Q000494:pharmacology; D000071999:S100 Calcium-Binding Protein A4; D009418:S100 Proteins / Q000378:metabolism
chemical_list: D005353:Fibronectins; D039661:Integrin beta3; D020574:Proto-Oncogene Proteins c-sis; D000071999:S100 Calcium-Binding Protein A4; D009418:S100 Proteins; C470944:S100a4 protein, mouse; D000077214:Becaplermin; D003094:Collagen; D051418:Focal Adhesion Kinase 2; C496053:Ptk2b protein, mouse