Collagenous stromal accumulations predict a worse clinical outcome in a variety of malignancies. Better tools are needed to elucidate the way in which collagen influences cancer cells. Here, we report a method to generate collagenous matrices that are deficient in key post-translational modifications and evaluate cancer cell behaviors on those matrices. We utilized genetic and biochemical approaches to inhibit lysine hydroxylation and glucosylation on collagen produced by MC-3T3-E1 murine osteoblasts (MC cells). Seeded onto MC cell-derived matrix surface, multicellular aggregates containing lung adenocarcinoma cells alone or in combination with cancer-associated fibroblasts dissociated with temporal and spatial patterns that were influenced by collagen modifications. These findings demonstrate the feasibility of generating defined collagen matrices that are suitable for cell culture studies.
Keywords: Co-culture models; Collagen; Collagen cross-links; DHLNL, Dehydro-dihydroxylysinonorleucine/its ketoamine; ER, Endoplasmic Reticulum; G, Galactosyl group; GG, Glucosylgalactosyl group; HLCCs, Hydroxylysine aldehyde-derived collagen cross-links; HLNL, Dehydro-hydroxylysinonorleucine/its ketoamine; Hyl, Hydroxylysine; Hylald, Aldehide Hydroxylysine; Hyp, Hydroxyproline; LCC, Lysine aldehyde–derived cross-links; LH, Lysyl hydroxylases; LOX, Lysyl oxidases; Lung cancer; Lys, Lysine; Lysald, Aldehide Lysine; Lysyl hydroxylases; Metastasis; PGGHG, Glucosylgalactosylhydroxylysine glucosidase; PTMs, Post-translational modifications; Pro, Proline; hLys, Helical domain Lysine; tLys, Telopeptidyl Lysine.
© 2020 The Authors.