Focal adhesion kinase is required for ß-catenin-induced mobilization of epidermal stem cells.

Focal adhesion kinase (FAK) is a non-receptor tyrosine kinase that integrates signals downstream of integrin and growth factor activation. Previously, we have shown that skin-specific loss of fak prevents chemically induced skin carcinogenesis in mice following phorbol ester treatment. In this study, we show that skin-specific deletion of fak prevents mobilization of stem cells within the bulge region of the hair follicle, which are the precursors of papillomas following phorbol ester treatment. We also show that phorbol ester treatment results in activation of-catenin within the skin and that FAK is required for ß-catenin-induced stem cell mobilization. In addition, inhibition of Src kinase activity, a major binding partner of FAK also prevents stem cell mobilization. We show that FAK is required for the nuclear localization of ß-catenin in the skin following phorbol ester treatment and the transcriptional activation of the ß-catenin target gene c-Myc. This provides the first evidence of cross-talk between integrin and Wnt signalling pathways in the control of epidermal stem cells and the early events associated with skin carcinogenesis.

The role of focal adhesion kinase catalytic activity on the proliferation and migration of squamous cell carcinoma cells.

Focal adhesion kinase (FAK) is upregulated in several epithelial tumours and there has been considerable interest in developing small molecule kinase inhibitors of FAK. However, FAK also has important adaptor functions within the cell, integrating signals from both integrins and growth factors. To investigate the role of FAKs kinase domain, we generated fak-deficient squamous cell carcinoma (SCC) cell lines. Re-expression of a wild type or kinase dead FAK allowed us to delineate its kinase dependent functions. In addition, we used the novel FAK kinase inhibitor PF-562,271. The kinase activity of FAK was important for tumour cell migration and polarity but more striking was its requirement for the anchorage independent 3 dimensional (3D) proliferation of SCC cells and their growth as xenografts in mice. Inhibition of FAK activity and prevention of growth in 3D correlated with Src inhibition. We further identified a mechanism whereby FAK regulates proliferation in 3D via regulation of the kinase activity of Src. This was dependent on the kinase activity of FAK and its resulting phosphorylation on Y397 that provides a high affinity binding site for Src. These data support the further development of FAK kinase inhibitors as agents that have the potential to inhibit both tumour cell migration and proliferation.

Diversity of matriptase expression level and function in breast cancer.

Overexpression of matriptase has been reported in a variety of human cancers and is sufficient to trigger tumor formation in mice, but the importance of matriptase in breast cancer remains unclear. We analysed matriptase expression in 16 human breast cancer cell lines and in 107 primary breast tumors. The data revealed considerable diversity in the expression level of this protein indicating that the significance of matriptase may vary from case to case. Matriptase protein expression was correlated with HER2 expression and highest expression was seen in HER2-positive cell lines, indicating a potential role in this subgroup. Stable overexpression of matriptase in two breast cancer cell lines had different consequences. In MDA-MB-231 human breast carcinoma cells the only noted consequence of matriptase overexpression was modestly impaired growth in vivo. In contrast, overexpression of matriptase in 4T1 mouse breast carcinoma cells resulted in visible changes in morphology, actin staining and cell to cell contacts. This correlated with downregulation of the cell-cell adhesion molecule E-cadherin. These results suggest that the functions of matriptase in breast cancer are likely to be variable and cell context dependent.

Telavancin: in vitro activity against staphylococci in a biofilm model.

OBJECTIVES: To assess the in vitro activity of the novel lipoglycopeptide telavancin against staphylococcal biofilms using an in vitro pharmacokinetic model. METHODS: Using the Sorbarod model, biofilms were established. The strains tested included methicillin-susceptible and -resistant strains of Staphylococcus aureus and coagulase-negative staphylococci, as well as glycopeptide-intermediate S. aureus (GISA). The biofilms were exposed to exponentially decreasing concentrations of telavancin and four comparator antibiotics, vancomycin, teicoplanin, linezolid and moxifloxacin and the bactericidal activity of the antibiotics was assessed. The concentrations of the antibiotics used in these experiments corresponded to peak serum levels achievable in humans and the rates at which drug concentrations were decreased corresponded to their elimination half-lives. RESULTS: All of the drugs tested produced a reduction in the number of bacteria eluted from the biofilms. Telavancin was more effective than the commercially available glycopeptides, vancomycin and teicoplanin, and of the three, was the most active agent against both the non-GISA and GISA strains. Of all the antibiotics tested, moxifloxacin produced the greatest reduction in biofilm cells, but only against the non-GISA strains. CONCLUSIONS: Telavancin exhibited substantial antimicrobial activity against staphylococcal biofilms, including GISA strains. This study supports the case for the evaluation of telavancin in the treatment of staphylococcal biofilm-associated infections.