title: Collaboration of 3D context and extracellular matrix in the development of glioma stemness in a 3D model.
abstract: A hierarchy of cellular stemness exists in certain cancers, and any successful strategy to treat such cancers would have to eliminate the self-renewing tumor-initiating cells at the apex of the hierarchy. The cellular microenvironment, in particular the extracellular matrix (ECM), is believed to have a role in regulating stemness. In this work, U251 glioblastoma cells are cultured on electrospun polystyrene (ESPS) scaffolds coated with an array of 7 laminin isoforms to provide a 3D model for stem cell-related genes and proteins expression studies. We observed collaboration between 3D context and laminins in promoting glioma stemness. Depending on the laminin isoform presented, U251 cells cultured on ESPS scaffolds (3D) exhibited increased expression of stemness markers compared to those cultured on tissue culture polystyrene (2D). Our results indicate the influence of 3D (versus 2D) context on integrin expression, specifically, the upregulation of the laminin-binding integrins alpha 6 and beta 4. By a colony forming assay, we showed enhanced clonogenicity of cells grown on ESPS scaffolds in collaboration with laminins 411, 421, 511 and 521. Evaluation of patient glioma databases demonstrated significant enrichment of integrin and ECM pathway networks in tumors of worse prognosis, consistent with our observations. The present results demonstrate how 3D versus 2D context profoundly affects ECM signaling, leading to stemness.
authors.affiliation: Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, The Nanos, Singapore 138669, Singapore.
authors.forename: Nina K L
authors.initials: NK
authors.lastname: Ma
authors.affiliation: Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, The Nanos, Singapore 138669, Singapore.
authors.forename: Jia Kai
authors.initials: JK
authors.lastname: Lim
authors.affiliation: Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, The Nanos, Singapore 138669, Singapore; School of Applied Science, Temasek Polytechnic, Singapore 529757, Singapore.
authors.forename: Meng Fatt
authors.initials: MF
authors.lastname: Leong
authors.affiliation: Department of Research, National Neuroscience Institute, Singapore 308433, Singapore; Singapore Institute for Clinical Sciences, Agency for Science, Technology and Research (A*STAR), Singapore 117609, Singapore; School of Biological Sciences, Nanyang Technological University, Singapore 637551, Singapore.
authors.forename: Edwin
authors.initials: E
authors.lastname: Sandanaraj
authors.affiliation: Singapore Institute for Clinical Sciences, Agency for Science, Technology and Research (A*STAR), Singapore 117609, Singapore; Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117597, Singapore; Department of Neurosurgery, National Neuroscience Institute, Singapore 308433, Singapore; Duke-National University of Singapore Graduate Medical School, Singapore 169857, Singapore.
authors.forename: Beng Ti
authors.initials: BT
authors.lastname: Ang
authors.affiliation: Department of Research, National Neuroscience Institute, Singapore 308433, Singapore; Duke-National University of Singapore Graduate Medical School, Singapore 169857, Singapore; Division of Medical Sciences, Humphrey Oei Institute of Cancer Research, National Cancer Centre, Singapore 169610, Singapore.
authors.forename: Carol
authors.initials: C
authors.lastname: Tang
authors.affiliation: Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, The Nanos, Singapore 138669, Singapore. Electronic address: awan@ibn.a-star.edu.sg.
authors.forename: Andrew C A
authors.initials: AC
authors.lastname: Wan
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000378:metabolism; D001932:Brain Neoplasms / Q000378:metabolism / Q000473:pathology*; D005109:Extracellular Matrix / Q000473:pathology*; D005910:Glioma / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D008855:Microscopy, Electron, Scanning; D008954:Models, Biological; D014411:Neoplastic Stem Cells / Q000473:pathology*
chemical_list: D018528:ATP-Binding Cassette Transporters
keywords: 3D context; Extracellular matrix; Glioma stemness; Integrins; Laminin isoforms