Alex Helkin1, Kristopher G Maier1, Vivian Gahtan2. 1. SUNY Upstate Medical University, Division of Vascular Surgery and Endovascular Services, Syracuse, NY, USA; Department of Veterans Affairs VA Healthcare Network Upstate New York at Syracuse, Syracuse, NY, USA. 2. SUNY Upstate Medical University, Division of Vascular Surgery and Endovascular Services, Syracuse, NY, USA; Department of Veterans Affairs VA Healthcare Network Upstate New York at Syracuse, Syracuse, NY, USA. Electronic address: gahtanv@upstate.edu.
Abstract
INTRODUCTION: The thrombospondins (TSPs) are matricellular proteins that exert multifunctional effects by binding cytokines, cell-surface receptors and other proteins. TSPs play important roles in vascular pathobiology and are all expressed in arterial lesions. The differential effects of TSP-1, -2, and -5 represent a gap in knowledge in vascular smooth muscle cell (VSMC) physiology. Our objective is to determine if structural differences of the TSPs imparted different effects on VSMC functions critical to the formation of neointimal hyperplasia. We hypothesize that TSP-1 and -2 induce similar patterns of migration, proliferation and gene expression, while the effects of TSP-5 are different. METHODS: Human aortic VSMC chemotaxis was tested for TSP-2 and TSP-5 (1-40 μg/mL), and compared to TSP-1 and serum-free media (SFM) using a modified Boyden chamber. Next, VSMCs were exposed to TSP-1, TSP-2 or TSP-5 (0.2-40 μg/mL). Proliferation was assessed by MTS assay. Finally, VSMCs were exposed to TSP-1, TSP-2, TSP-5 or SFM for 3, 6 or 24 h. Quantitative real-time PCR was performed on 96 genes using a microfluidic card. Statistical analysis was performed by ANOVA or t-test, with p < 0.05 being significant. RESULTS: TSP-1, TSP-2 and TSP-5 at 20 μg/mL all induce chemotaxis 3.1 fold compared to serum-free media. TSP-1 and TSP-2 induced proliferation 53% and 54% respectively, whereas TSP-5 did not. In the gene analysis, overall, cardiovascular system development and function is the canonical pathway most influenced by TSP treatment, and includes multiple growth factors, cytokines and proteases implicated in cellular migration, proliferation, vasculogenesis, apoptosis and inflammation pathways. CONCLUSIONS AND RELEVANCE: The results of this study indicate TSP-1, -2, and -5 play active roles in VSMC physiology and gene expression. Similarly to TSP-1, VSMC chemotaxis to TSP-2 and -5 is dose-dependent. TSP-1 and -2 induces VSMC proliferation, but TSP-5 does not, likely due conservation of N-terminal domains in TSP-1 and -2. In addition, TSP-1, -2 and -5 significantly affect VSMC gene expression; however, little overlap exists in the specific genes altered. This study further delineates TSP-1, -2 and -5's contributions to processes related to VSMC physiology. Published by Elsevier Inc.
INTRODUCTION: The thrombospondins (TSPs) are matricellular proteins that exert multifunctional effects by binding cytokines, cell-surface receptors and other proteins. TSPs play important roles in vascular pathobiology and are all expressed in arterial lesions. The differential effects of TSP-1, -2, and -5 represent a gap in knowledge in vascular smooth muscle cell (VSMC) physiology. Our objective is to determine if structural differences of the TSPs imparted different effects on VSMC functions critical to the formation of neointimal hyperplasia. We hypothesize that TSP-1 and -2 induce similar patterns of migration, proliferation and gene expression, while the effects of TSP-5 are different. METHODS:Human aortic VSMC chemotaxis was tested for TSP-2 and TSP-5 (1-40 μg/mL), and compared to TSP-1 and serum-free media (SFM) using a modified Boyden chamber. Next, VSMCs were exposed to TSP-1, TSP-2 or TSP-5 (0.2-40 μg/mL). Proliferation was assessed by MTS assay. Finally, VSMCs were exposed to TSP-1, TSP-2, TSP-5 or SFM for 3, 6 or 24 h. Quantitative real-time PCR was performed on 96 genes using a microfluidic card. Statistical analysis was performed by ANOVA or t-test, with p < 0.05 being significant. RESULTS:TSP-1, TSP-2 and TSP-5 at 20 μg/mL all induce chemotaxis 3.1 fold compared to serum-free media. TSP-1 and TSP-2 induced proliferation 53% and 54% respectively, whereas TSP-5 did not. In the gene analysis, overall, cardiovascular system development and function is the canonical pathway most influenced by TSP treatment, and includes multiple growth factors, cytokines and proteases implicated in cellular migration, proliferation, vasculogenesis, apoptosis and inflammation pathways. CONCLUSIONS AND RELEVANCE: The results of this study indicate TSP-1, -2, and -5 play active roles in VSMC physiology and gene expression. Similarly to TSP-1, VSMC chemotaxis to TSP-2 and -5 is dose-dependent. TSP-1 and -2 induces VSMC proliferation, but TSP-5 does not, likely due conservation of N-terminal domains in TSP-1 and -2. In addition, TSP-1, -2 and -5 significantly affect VSMC gene expression; however, little overlap exists in the specific genes altered. This study further delineates TSP-1, -2 and -5's contributions to processes related to VSMC physiology. Published by Elsevier Inc.
Authors: J C Rodriguez-Manzaneque; T F Lane; M A Ortega; R O Hynes; J Lawler; M L Iruela-Arispe Journal: Proc Natl Acad Sci U S A Date: 2001-10-16 Impact factor: 11.205
Authors: Natasha M Rogers; Maryam Sharifi-Sanjani; Mingyi Yao; Kedar Ghimire; Raquel Bienes-Martinez; Stephanie M Mutchler; Heather E Knupp; Jeffrey Baust; Enrico M Novelli; Mark Ross; Claudette St Croix; Johannes C Kutten; Caitlin A Czajka; John C Sembrat; Mauricio Rojas; David Labrousse-Arias; Timothy N Bachman; Rebecca R Vanderpool; Brian S Zuckerbraun; Hunter C Champion; Ana L Mora; Adam C Straub; Richard A Bilonick; Maria J Calzada; Jeffrey S Isenberg Journal: Cardiovasc Res Date: 2016-10-13 Impact factor: 10.787
Authors: Thomas C F Bodewes; Joel M Johnson; Michael Auster; Cindy Huynh; Sriya Muralidharan; Mauricio Contreras; Frank W LoGerfo; Leena Pradhan-Nabzdyk Journal: FASEB J Date: 2016-09-26 Impact factor: 5.191
Authors: Emma Plana; Laura Gálvez; Pilar Medina; Silvia Navarro; Victoria Fornés-Ferrer; Joaquín Panadero; Manuel Miralles Journal: Int J Mol Sci Date: 2020-06-28 Impact factor: 5.923