title: Multifunctional bioscaffolds for 3D culture of melanoma cells reveal increased MMP activity and migration with BRAF kinase inhibition.
abstract: Matrix metalloproteinases (MMPs) are important for many different types of cancer-related processes, including metastasis. Understanding the functional impact of changes in MMP activity during cancer treatment is an important facet not typically evaluated as part of preclinical research. With MMP activity being a critical component of the metastatic cascade, we designed a 3D hydrogel system to probe whether pharmacological inhibition affected human melanoma cell proteolytic activity; metastatic melanoma is a highly aggressive and drug-resistant form of skin cancer. The relationship between MMP activity and drug treatment is unknown, and therefore we used an in situ fluorogenic MMP sensor peptide to determine how drug treatment affects melanoma cell MMP activity in three dimensions. We encapsulated melanoma cells from varying stages of progression within PEG-based hydrogels to examine the relationship between drug treatment and MMP activity. From these results, a metastatic melanoma cell line (A375) and two inhibitors that inhibit RAF (PLX4032 and sorafenib) were studied further to determine whether changes in MMP activity led to a functional change in cell behavior. A375 cells exhibited increased MMP activity despite an overall decrease in metabolic activity with PLX4032 treatment. The changes in proteolytic activity correlated with increased cell elongation and increased single-cell migration. In contrast, sorafenib did not alter MMP activity or cell motility, showing that the changes induced by PLX4032 were not a universal response to small-molecule inhibition. Therefore, we argue the importance of studying MMP activity with drug treatment and its possible implications for unwanted side effects.
authors: Department of Chemical and Biological Engineering, BioFrontiers Institute, and Howard Hughes Medical Institute, University of Colorado Boulder, Boulder, CO 80309; and Department of Biomedical Engineering and Comprehensive Cancer Center, The Ohio State University, Columbus, OH 43210 leight.1@osu.edu kristi.anseth@colorado.edu.
authors: Jennifer L
authors: JL
authors: Leight
authors: Department of Chemical and Biological Engineering, BioFrontiers Institute, and.
authors: Emi Y
authors: EY
authors: Tokuda
authors: Department of Chemical and Biological Engineering, BioFrontiers Institute, and.
authors: Caitlin E
authors: CE
authors: Jones
authors: Department of Chemical and Biological Engineering, BioFrontiers Institute, and.
authors: Austin J
authors: AJ
authors: Lin
authors: Department of Chemical and Biological Engineering, BioFrontiers Institute, and Howard Hughes Medical Institute, University of Colorado Boulder, Boulder, CO 80309; and leight.1@osu.edu kristi.anseth@colorado.edu.
authors: Kristi S
authors: KS
authors: Anseth
mesh_terms: D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects*; D017364:Collagenases / Q000378:metabolism*; D006801:Humans; D020100:Hydrogels / Q000737:chemistry; D007211:Indoles / Q000494:pharmacology*; D008545:Melanoma / Q000201:enzymology* / Q000473:pathology; D009536:Niacinamide / Q000031:analogs & derivatives* / Q000494:pharmacology; D010671:Phenylurea Compounds / Q000494:pharmacology*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D048493:Proto-Oncogene Proteins B-raf / Q000037:antagonists & inhibitors* / Q000378:metabolism; D000077157:Sorafenib; D013449:Sulfonamides / Q000494:pharmacology*; D054457:Tissue Scaffolds / Q000737:chemistry*; D000077484:Vemurafenib
chemical_list: D020100:Hydrogels; D007211:Indoles; D010671:Phenylurea Compounds; D047428:Protein Kinase Inhibitors; D013449:Sulfonamides; D000077484:Vemurafenib; D009536:Niacinamide; D000077157:Sorafenib; C482119:BRAF protein, human; D048493:Proto-Oncogene Proteins B-raf; D017364:Collagenases
keywords: BRAF/MEK; cancer; cell motility; hydrogel; matrix metalloproteinase