Glycated collagen – a 3D matrix system to study pathological cell behavior

Abstract

A hyperglycemic condition like diabetes in patients renders them with an increased risk of developing breast cancer. Hyperglycemia and ageing increase the non-enzymatic glycosylation (glycation) of nearly all proteins in our body including collagen type I, which is an important extracellular matrix (ECM) component. This results in the formation of advanced glycated end products (AGEs), which can form covalent crosslinks in collagen fibers and change the overall architecture and stiffness of the matrix. In this study we have used MDA-MB-231 breast cancer cells to study the interaction of tumor cells with glycated collagen and have explored the role of matrix architecture and RAGE-mediated signaling in cellular behavior. We mimicked the non-enzymatic glycation of protein by treating collagen I with glucose or ribose and found that crosslinking due to AGEs induces collagen fiber bundling and an increase in pore size and stiffness of the matrix. We also observed that AGE formation triggers AGE–RAGE signaling playing a role in the morphology and migration of cells. Furthermore, our study suggests an interplay of the pore size of the collagen matrix and RAGE mediated signaling in 3D invasion of cells and our findings demonstrate that the effect of the AGE–RAGE interaction is more pronounced than that of an altered matrix architecture. This study has helped us develop a 3D system using glycated collagen to study the effects of pathological conditions such as diabetes on extracellular matrix proteins, which may have downstream effects on cell behavior and dysfunction.