Type III and V collagens modulate the expression and assembly of EDA+ fibronectin in the extracellular matrix of defective Ehlers-Danlos syndrome fibroblasts

Background: Alternative splicing of EDA fibronectin (FN) region is a cell type- and development-regulated
mechanism controlled by pathological processes, growth factors and extracellular matrix (ECM). Classic
and vascular Ehlers–Danlos syndrome (cEDS and vEDS) are connective tissue disorders caused by COL5A1/
COL5A2 and COL3A1 gene mutations, leading to an in vivo abnormal collagen fibrillogenesis and to an in
vitro defective organisation in the ECM of type V (COLLV) and type III collagen (COLLIII). These defects induce
the FN-ECM disarray and the decrease of COLLs and FN receptors, the α2β1 and α5β1 integrins. Purified
COLLV and COLLIII restore the COLL-FN-ECMs in both EDS cell strains.
Methods: Real-time PCR, immunofluorescence microscopy, andWestern blotting were used to investigate the
effects of COLLs on FN1 gene expression, EDA region alternative splicing, EDA+-FN-ECM assembly, α5β1
integrin and EDA+-FN-specific α9 integrin subunit organisation, α5β1 integrin and FAK co-regulation in
EDS fibroblasts.
Results: COLLV-treated cEDS and COLLIII-treated vEDS fibroblasts up-regulate the FN1 gene expression, mod-
ulate the EDA+ mRNA maturation and increase the EDA+-FN levels, thus restoring a control-like FN-ECM,
which elicits the EDA+-FN-specific α9β1 integrin organisation, recruits the α5β1 integrin and switches on
the FAK binding and phosphorylation.
Conclusion: COLLs regulate the EDA+-FN-ECM organisation at transcriptional and post-transcriptional level
and activate the α5β1–FAK complexes. COLLs also recruit the α9β1 integrin involved in the assembly of
the EDA+-FN-ECM in EDS cells.
General significance: The knowledge of the COLLs-ECM role in FN isotype expression and in EDA+-FN-ECM-
mediated signal transduction adds insights in the ECM remodelling mechanisms in EDS cells.
© 2012 Published by Elsevier B.V. 42
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