A8.15 The Focal Contact Protein Lasp-1 Modulates the Migration Capacity of Synovial Fibroblasts. (25th February 2013)

• Record Type: Journal Article
• Authors: Korb-Pap, Adelheid; Beckmann, Denise; Hillen, Jan; Heitzmann, Marianne; Chew, Catherine S; Butz, Stefan; Vestweber, Dietmar; Pavenstädt, Hermann; Pap, Thomas
• Is Part Of: Annals of the rheumatic diseases. Volume 72:Supplement 1(2013)
• Page Start: A62
• Page End: A63
• Publication Date: 2013-02-25
• Subjects: Rheumatism -- Periodicals; 616.723005
• DOI: 10.1136/annrheumdis-2013-203222.15
• Languages: English
• ISSNs: 0003-4967

Abstract

Abstract : Background and Objectives: RA synovial fibroblasts (SF) have been suggested to contribute to the spreading of disease through their ability to leave cartilage destruction sites, migrate via the bloodstream and re-initiate the destructive process at distant articular cartilage surfaces. In this context, the actin-crosslinking protein Lasp-1 is of interest, because it is localised at leading edges of migrating cells and regulates metastatic dissemination of different tumours. Therefore, it is particularly important to investigate the role of Lasp-1 in SF migration and its effects on RA. Materials and Methods: To identify different Lasp-1 expression levels in the hind paws of wt and hTNFtg mice, an established model for human RA, Western- blot analyses were performed. In parallel, Lasp-1 expression and its sub-cellular distribution was investigated in SF from wt and hTNFtg mice by Western-blot analyses and immunofluorescence. The migratory capacity of SFs derived from wild-type, Lasp-1 -/-, hTNFtg and Lasp1 -/- /hTNFtg mice was studied in a modified scratch assay as well as in live cell imaging studies. Furthermore, a transmigration assay using SF from all four genotypes and murine endothelioma cells (bEnd.5) as an endothelial barrier was carried out. For more detailed information, SF transmigration was evaluated when endothelial cells were also pre-treated with TNF-alpha, mimicking inflammatory conditions. Results: Lasp-1 expression is upregulated in SF from hTNFtg mice and localises to structures of cell adhesion and invasion. In the scratch assay, a significantly reduced migration rate was detected in Lasp-1 -/- SFs after 24 hrs (–43.7% versus wt, p < 0.05) and in Lasp1 -/- /hTNFtg, respectively (–69.11% versus hTNFtg, p < 0.05). Live cell imaging studies showed a slower migration and striking differences in migration morphology of Lasp1 -/- /hTNFtg compared to hTNFtg SF. Furthermore, analyses showed a significant reduction of transmigration of Lasp1 -/- /hTNFtg compared to hTNFtg SF that was even enhanced by TNF-alpha stimulation of the endothelial cells. Interestingly, interbred Lasp1 -/- /hTNFtg mice presented milder clinical symptoms and analyses of histopathology revealed less cartilage degradation and less attachment of synovial tissue to the cartilage than hTNFtg mice at an age of 14 weeks. Conclusions: Our data provide that the migratory capacity of SF is regulated by Lasp-1 and influences the severity of arthritis in hTNFtg mice. SF – when activated – migrate through the formation of invasive and adhesive membrane structures such as invadopodia, where Lasp-1 is prominently localised. Thus, targeting Lasp-1 may be a promising strategy to reduce the invasive and migratory behaviour of synovial fibroblasts in RA.