Wei Wang1, Nathalie Burg1, Spandana Vootukuri1, Barry S Coller2. 1. Allen and Frances Adler Laboratory of Blood and Vascular Biology, The Rockefeller University, New York, NY, United States. 2. Allen and Frances Adler Laboratory of Blood and Vascular Biology, The Rockefeller University, New York, NY, United States. Electronic address: collerb@rockefeller.edu.
Abstract
BACKGROUND: Transforming growth factor-β1 (TGF-β1) has been implicated in the pathogenesis of aortic valve stenosis (AS). There is, however, little direct evidence for a role of active TGF-β1 in AS due to the sensitivity of current assays. We searched for evidence of plasma TGF-β1 activation by assaying Smad2/3 phosphorylation in circulating leukocytes and platelet-leukocyte aggregates (PLAs) in a mouse model of AS (Reversa). METHODS: Echocardiography was used to measure AS and cardiac function. Intracellular phospho-flow cytometry in combination with optical fluorescence microscopy was used to detect PLAs and p-Smad2/3 levels. RESULTS: Reversa mice on a western diet developed AS, had significantly increased numbers of PLAs and more intense staining for p-Smad2/3 in both PLAs and single leukocytes (all p<0.05). p-Smad2/3 staining was more intense in PLAs than in single leukocytes in both diet groups (p<0.05) and correlated with plasma total TGF-β1 levels (r=0.38, p=0.05 for PLAs and r=0.37, p=0.06 for single leukocytes) and reductions in ejection fraction (r=-0.42, p=0.03 for PLAs and r=-0.37, p=0.06 for single leukocytes). CONCLUSIONS: p-Smad2/3 staining is more intense in leukocytes of hypercholesterolemic mice that developed AS, suggesting increased circulating active TGF-β1 levels. Leukocyte p-Smad2/3 may be a valuable surrogate indicator of circulating active TGF-β1.
BACKGROUND: Transforming growth factor-β1 (TGF-β1) has been implicated in the pathogenesis of aortic valve stenosis (AS). There is, however, little direct evidence for a role of active TGF-β1 in AS due to the sensitivity of current assays. We searched for evidence of plasma TGF-β1 activation by assaying Smad2/3 phosphorylation in circulating leukocytes and platelet-leukocyte aggregates (PLAs) in a mouse model of AS (Reversa). METHODS: Echocardiography was used to measure AS and cardiac function. Intracellular phospho-flow cytometry in combination with optical fluorescence microscopy was used to detect PLAs and p-Smad2/3 levels. RESULTS: Reversa mice on a western diet developed AS, had significantly increased numbers of PLAs and more intense staining for p-Smad2/3 in both PLAs and single leukocytes (all p<0.05). p-Smad2/3 staining was more intense in PLAs than in single leukocytes in both diet groups (p<0.05) and correlated with plasma total TGF-β1 levels (r=0.38, p=0.05 for PLAs and r=0.37, p=0.06 for single leukocytes) and reductions in ejection fraction (r=-0.42, p=0.03 for PLAs and r=-0.37, p=0.06 for single leukocytes). CONCLUSIONS: p-Smad2/3 staining is more intense in leukocytes of hypercholesterolemicmice that developed AS, suggesting increased circulating active TGF-β1 levels. Leukocyte p-Smad2/3 may be a valuable surrogate indicator of circulating active TGF-β1.
Authors: Roza Badr Eslam; Thomas Gremmel; Alexandra Schneller; Michael Stegfellner; Alexandra Kaider; Christine Mannhalter; Irene Lang; Simon Panzer Journal: Thromb Res Date: 2011-08-03 Impact factor: 3.944
Authors: L M Wakefield; T S Winokur; R S Hollands; K Christopherson; A D Levinson; M B Sporn Journal: J Clin Invest Date: 1990-12 Impact factor: 14.808
Authors: Jasimuddin Ahamed; Nathalie Burg; Keiji Yoshinaga; Christin A Janczak; Daniel B Rifkin; Barry S Coller Journal: Blood Date: 2008-06-10 Impact factor: 22.113
Authors: Alexander Meyer; Wei Wang; Jiaxiang Qu; Lori Croft; Jay L Degen; Barry S Coller; Jasimuddin Ahamed Journal: Blood Date: 2011-12-01 Impact factor: 25.476