Cellular fibronectin toolset

Expression vectors encoding human cellular fibronectin variants harboring the alternatively spliced Extra Domains B and/or A (or neither) ; fibronectin variant-expressing cells and purified recombinant cellular fibronectin variant proteins.

Interest / Relevance: Cellular (also known as oncofetal) fibronectin variants harboring one or both alternatively spliced so-called extra domains (EDB and EDA) play a central bioregulatory role during development, repair processes and fibrosis.

Keywords: extracellular matrix, fibrosis, angiogenesis, tumor microenvironment

Scientist's name: Ellen VAN OBBERGHEN-SCHILLING
Georgios Efthymiou
Agata Radwanska

Publications:

Efthymiou G*, Radwanska A*, Grapa A-I, Beghelli-de la Forest Divonne S, Grall D, Schaub S, Hattab M, Pisano S, Poet M, Pisani D, Counillon L, Descombes X, Blanc-Feraud L and Van Obberghen-Schilling E. Fibronectin Extra Domains tune cellular responses and confer topographically distinct features to fibril networks. J Cell Sci. 2021 : jcs.252957 doi: 10.1242/jcs.252957 Published 1 February 2021.

Efthymiou G*, Saint A, Ruff M, Rekad Z, Ciais D and Van Obberghen-Schilling E Shaping Up the Tumor Microenvironment with Cellular Fibronectin. Front Oncol. 2020 Apr 30,10:641. doi: 10.3389/fonc.2020.00641.

Soubies E, Radwanska A, Grall D, Aubert GA, Blanc-Feraud L, Van Obberghen-Schilling E and Schaub, S. Nanometric axial resolution of fibronectin assembly achieved with an efficient reconstruction approach for multi-angle-TIRF microscopy Sci Rep. 2019 Feb 13,9(1):1926. doi: 10.1038/s41598-018-36119-3.

Grapa AI, Meunier R, Blanc-Féraud L, Efthymiou G, Schaub S, Radwanska A, Van Obberghen-Schilling E and Descombes X. (2018) Classification of fibronectin variants with curvelets IEEE International Symposium on Biomedical Imaging

Radwanska A, Grall D, Schaub S, Beghelli-de la Forest Divonne S, Ciai, D, Sudaka A, Rekima S, Rupp T, Orend G and Van Obberghen-Schilling E. (2017) Counterbalancing anti-adhesive effects of Tenascin-C through fibronectin expression in endothelial cells. Sci Rep. Oct 6,7(1):12762. doi: 10.1038/s41598-017-13008-9.

Gopal S*, Veracini L*, Grall D, Butori C, Schaub S, Audebert S, Camoin L, Baudelet E, Radwanska A, Beghelli-de la Forest Divonne S, Violette SM, Weinreb PH, Rekima S, Ilie M, Sudaka A, Hofman P and Van Obberghen-Schilling E. Fibronectin-guided migration of carcinoma collectives. Nat Commun. 2017 Jan 19,8:14105. doi: 10.1038
comms14105

Serres E., Debarbieux F.*, Stanchi F.*, Maggiorella L., Grall D., Rougon G., and Van Obberghen-Schilling E. (2014) Fibronectin expression in glioblastomas promotes cell cohesion, collective invasion of basement membrane in vitro and orthotopic tumor growth in mice. Oncogene 33:3451-62.

Cseh B, Fernandez-Sauze S, Grall D, Schaub S, Doma E, and Van Obberghen-Schilling E. Autocrine fibronectin directs matrix assembly and crosstalk between cell-matrix and cell-cell adhesion in vascular endothelial cells. J Cell Sci. (2010) 123, 3989-3999.

Detailed technology overview: Detailed technology is described in :
Efthymiou G, et al. Fibronectin Extra Domains tune cellular responses and confer topographically distinct features to fibril networks. J Cell Sci. 2021 : jcs.252957 doi: 10.1242/jcs.252957 Published 1 February 2021.

- Construction of lentiviral expression vectors harboring the alternatively-spliced « cellular FN » variants containing Extra Domain A and/or B, or no Extra Domain (plasma-like FN)
- Production of lentiviral particles for transduction of FN variant-expressing cells
- Expression of FN variants in (assembly-incompetent) HEK293 cells
- Purification of soluble FN variants secreted by FN-expressing HEK293 cells
- Expression of FN variants in assembly-competent cells
- Addition of purified FN variants to cells for functional studies
- Preparation of variant-specific « cell-derived matrices »

Reference:

RT00508