The 1ALCTL and 1BLCTL isoforms of Arg/Abl2 induce fibroblast activation and extra cellular matrix remodelling differently.

Barbara Torsello,Sofia De Marco,Roberto A Perego,Roberto Giovannoni,Roberta Corti,Silvia Bombelli,Elisa Chisci,Cristina Bianchi,Valeria Cassina,Davide Bernasconi

doi:10.1242/bio.038554

Barbara Torsello, Sofia De Marco + Show 8 more

Open Access

https://doi.org/10.1242/bio.038554

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Journal: Biology open	Publication Date: Mar 15, 2019
Citations: 7	License type: CC BY 4.0

Affiliation: University of Milano-Bicocca, University of Milan

Abstract

ABSTRACTThe fibrotic tissue and the stroma adjacent to cancer cells are characterised by the presence of activated fibroblasts (myofibroblasts) which play a role in creating a supportive tissue characterised by abundant extracellular matrix (ECM) secretion. The myofibroblasts remodel this tissue through secreted molecules and modulation of their cytoskeleton and specialized contractile structures. The non-receptor protein tyrosine kinase Arg (also called Abl2) has the unique ability to bind directly to the actin cytoskeleton, transducing diverse extracellular signals into cytoskeletal rearrangements. In this study we analysed the 1ALCTL and 1BLCTL Arg isoforms in Arg−/− murine embryonal fibroblasts (MEF) cell line, focusing on their capacity to activate fibroblasts and to remodel ECM. The results obtained showed that Arg isoform 1BLCTL has a major role in proliferation, migration/invasion of MEF and in inducing a milieu able to modulate tumour cell morphology, while 1ALCTL isoform has a role in MEF adhesion maintaining active focal adhesions. On the whole, the presence of Arg in MEF supports the proliferation, activation, adhesion, ECM contraction and stiffness, while the absence of Arg affected these myofibroblast features.This article has an associated First Person interview with the first author of the paper.

Highlights

The fibrotic process, after a chronic injury, results in an excessive scar tissue deposition and development of fibrosis, and has in the activated fibroblasts the main players (Yazdani et al, 2017)
Stable expression and tyrosine kinase activity of 1ALCTL and 1BLCTL Arg isoforms transfected in Arg−/− murine embryonal fibroblasts (MEF) To study the role of 1ALCTL and 1BLCTL Arg isoforms in fibroblast activation, we cloned into the stable expression vector pCX-C1-EGFP plasmid (Cinti et al, 2015) the corresponding Arg cDNA sequences
A characteristic of activated fibroblasts is the high proliferation rate (Barron and Rowley, 2012; Li et al, 2016), we evaluated the effect of Arg isoforms on MEF proliferation counting the viable cells at different time points

Summary

Introduction

The fibrotic process, after a chronic injury, results in an excessive scar tissue deposition and development of fibrosis, and has in the activated fibroblasts (myofibroblasts) the main players (Yazdani et al, 2017). The stroma adjacent to cancer cells is a permissive and supportive environment (Mueller and Fusenig, 2004) in which there is a variable presence of myofibroblasts, . CAF promote matrix remodelling by soluble factors (Kalluri and Zeisberg, 2006) and by the generation of tracks that enable the collective invasion of the tumour cells. Remodelling of the ECM and promotion of cancer cell invasion requires myofibroblast cytoskeletal rearrangement through contraction of the actomyosin cytoskeleton assembled in stress fibres (Barron and Rowley, 2012; Calvo et al, 2013)

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