Biglycan expression in the melanoma microenvironment promotes invasiveness via increased tissue stiffness inducing integrin-ß1 expression.

Novel targeted and immunotherapeutic approaches have revolutionized the treatment of metastatic melanoma. A better understanding of the melanoma-microenvironment, in particular the interaction of cells with extracellular matrix molecules, may help to further improve these new therapeutic strategies.We observed that the extracellular matrix molecule biglycan (Bgn) was expressed in certain human melanoma cells and primary fibroblasts when evaluated by microarray-based gene expression analysis. Bgn expression in the melanoma tissues correlated with low overall-survival and low progression-free-survival in patients. To understand the functional role of Bgn we used gene-targeted mice lacking functional Bgn. Here we observed that melanoma growth, metastasis-formation and tumor-related death were reduced in Bgn-/- mice compared to Bgn+/+ mice. In vitro invasion of melanoma cells into organotypic-matrices derived from Bgn-/- fibroblasts was reduced compared to melanoma invasion into Bgn-proficient matrices. Tissue stiffness as determined by atomic-force-microscopy was reduced in Bgn-/- matrices. Isolation of melanoma cells and fibroblasts from the stiffer Bgn+/+ matrices revealed an increase in integrin-ß1 expression compared to the Bgn-/- fibroblast matrices. Overexpression of integrin-ß1 in B16-melanoma cells abolished the survival benefit seen in Bgn-/- mice. Consistent with the studies performed in mice, the abundance of Bgn-expression in human melanoma samples positively correlated with the expression of integrin-ß1, which is in agreement with results from the organotypic invasion-assay and the in vivo mouse studies.This study describes a novel role for Bgn-related tissue stiffness in the melanoma-microenvironment via regulation of integrin-ß1 expression by melanoma cells in both mice and humans.

Mechanical load and mechanical integrity of lung cells - experimental mechanostimulation of epithelial cell- and fibroblast-monolayers.

Experimental mechanostimulation of soft biologic tissue is widely used to investigate cellular responses to mechanical stress or strain. Reactions on mechanostimulation are investigated in terms of morphological changes, inflammatory responses and apoptosis/necrosis induction on a cellular level. In this context, the analysis of the mechanical characteristics of cell-layers might allow to indicate patho-physiological changes in the cell-cell contacts. Recently, we described a device for experimental mechanostimulation that allows simultaneous measurement of the mechanical characteristics of cell-monolayers. Here, we investigated how cultivated lung epithelial cell- and fibroblast-monolayers behave mechanically under different amplitudes of biaxial distension. The cell monolayers were sinusoidally deflected to 5%, 10% or 20% surface gain and their mechanical properties during mechanostimulation were analyzed. With increasing stimulation amplitudes more pronounced reductions of cell junctions were observed. These findings were accompanied by a substantial loss of monolayer rigidity. Pulmonary fibroblast monolayers were initially stiffer but were stronger effected by the mechanostimulation compared to epithelial cell-monolayers. We conclude that, according to their biomechanical function within the pulmonary tissue, epithelial cells and fibroblasts differ with respect to their mechanical characteristics and tolerance of mechanical load.