Galectin-1 Restricts Vascular Smooth Muscle Cell Motility Via Modulating Adhesion Force and Focal Adhesion Dynamics

Min-Shao Tsai,Po-Chiao Chang,Lee-Young Chau,Yi-Ru Li,Fu-Tong Liu,Chih-Wen Yang,Heng-Huei Lin,Ing-Shouh Hwang,Kuo-I Lin,Ming-Tsai Chiang,Chiao-Po Hsu,Dong-Lin Tsai,Hsien-San Hou,Huan-Yuan Chen,Pei-Kuen Wei

doi:10.1038/s41598-018-29843-3

Abstract

Vascular smooth muscle cell (VSMC) migration play a key role in the development of intimal hyperplasia and atherosclerosis. Galectin-1 (Gal-1) is a redox-sensitive β-galactoside-binding lectin expressed in VSMCs with intracellular and extracellular localizations. Here we show that VSMCs deficient in Gal-1 (Gal-1-KO) exhibited greater motility than wild type (WT) cells. Likewise, Gal-1-KO-VSMC migration was inhibited by a redox-insensitive but activity-preserved Gal-1 (CSGal-1) in a glycan-dependent manner. Gal-1-KO-VSMCs adhered slower than WT cells on fibronectin. Cell spreading and focal adhesion (FA) formation examined by phalloidin and vinculin staining were less in Gal-1-KO-VSMCs. Concomitantly, FA kinase (FAK) phosphorylation was induced to a lower extent in Gal-1-KO cells. Analysis of FA dynamics by nocodazole washout assay demonstrated that FA disassembly, correlated with FAK de-phosphorylation, was faster in Gal-1-KO-VSMCs. Surface plasmon resonance assay demonstrated that CSGal-1 interacted with α5β1integrin and fibronectin in a glycan-dependent manner. Chemical crosslinking experiment and atomic force microscopy further revealed the involvement of extracellular Gal-1 in strengthening VSMC-fibronectin interaction. In vivo experiment showed that carotid ligation-induced neointimal hyperplasia was more severe in Gal-1-KO mice than WT counterparts. Collectively, these data disclose that Gal-1 restricts VSMC migration by modulating cell-matrix interaction and focal adhesion turnover, which limits neointimal formation post vascular injury.

Highlights

Galectins are a family of β-galactoside-binding proteins with a wide spectrum of biological functions in a variety of cell types and tissues[3]
In the present study we assessed the impacts of Gal-1 deficiency on platelet-derived growth factor (PDGF)-induced VSMC growth and migration responses using primary VSMCs isolated from wild type (WT) and Gal-1-knockout (KO) mice
To investigate whether Gal-1 has an impact on VSMC phenotype, we cultured primary VSMCs isolated from WT and Gal-KO mice and compared their growth rates and motilities in vitro

Summary

Introduction

Galectins are a family of β-galactoside-binding proteins with a wide spectrum of biological functions in a variety of cell types and tissues[3]. These proteins exhibit both extracellular and intracellular localizations[3,4]. Given that Gal-1 is a multifunctional protein, the potential roles of endogenous Gal-1 involved during VSMC remodeling post vascular injury deserve to be explored To this end, in the present study we assessed the impacts of Gal-1 deficiency on platelet-derived growth factor (PDGF)-induced VSMC growth and migration responses using primary VSMCs isolated from wild type (WT) and Gal-1-knockout (KO) mice. We assessed the impact of Gal-1 deficiency on the development of intima hyperplasia in animal model

Methods

Results

Conclusion