title: A three-dimensional engineered tumour for spatial snapshot analysis of cell metabolism and phenotype in hypoxic gradients.
abstract: The profound metabolic reprogramming that occurs in cancer cells has been investigated primarily in two-dimensional cell cultures, which fail to recapitulate spatial aspects of cell-to-cell interactions as well as tissue gradients present in three-dimensional tumours. Here, we describe an engineered model to assemble three-dimensional tumours by rolling a scaffold-tumour composite strip. By unrolling the strip, the model can be rapidly disassembled for snapshot analysis, allowing spatial mapping of cell metabolism in concert with cell phenotype. We also show that the establishment of oxygen gradients within samples that are shaped by oxygen-dependent signalling pathways, as well as the consequential variations in cell growth, response to hypoxic gradients extending from normoxia to severe hypoxia, and therapy responsiveness, are consistent with those of tumours in vivo. Moreover, by using liquid chromatography tandem mass spectrometry, we mapped cellular metabolism and identified spatially defined metabolic signatures of cancer cells to reveal both known and novel metabolic responses to hypoxia.
authors.affiliation: University of Toronto, Department of Chemical Engineering and Applied Chemistry, 200 College Street Toronto, Ontario M5S 3E5, Canada.
authors.forename: Darren
authors.initials: D
authors.lastname: Rodenhizer
authors.affiliation: MRC Cancer Unit, University of Cambridge, Hutchison/MRC Research Centre, Box 197, Cambridge Biomedical Campus, Cambridge CB2 0XZ, UK.
authors.forename: Edoardo
authors.initials: E
authors.lastname: Gaude
authors.affiliation: Princess Margaret Cancer Centre and Campbell Family Institute for Cancer Research, Departments of Medical Biophysics and Radiation Oncology, University Health Network, Toronto, Ontario M5G 2M9, Canada.
authors.forename: Dan
authors.initials: D
authors.lastname: Cojocari
authors.affiliation: University of Toronto, Department of Chemical Engineering and Applied Chemistry, 200 College Street Toronto, Ontario M5S 3E5, Canada.|Institute of Biomaterials and Biomedical Engineering, University of Toronto, 164 College Street, Toronto, Ontario M5S 3G9, Canada.
authors.forename: Radhakrishnan
authors.initials: R
authors.lastname: Mahadevan
authors.affiliation: MRC Cancer Unit, University of Cambridge, Hutchison/MRC Research Centre, Box 197, Cambridge Biomedical Campus, Cambridge CB2 0XZ, UK.
authors.forename: Christian
authors.initials: C
authors.lastname: Frezza
authors.affiliation: Princess Margaret Cancer Centre and Campbell Family Institute for Cancer Research, Departments of Medical Biophysics and Radiation Oncology, University Health Network, Toronto, Ontario M5G 2M9, Canada.
authors.forename: Bradly G
authors.initials: BG
authors.lastname: Wouters
authors.affiliation: University of Toronto, Department of Chemical Engineering and Applied Chemistry, 200 College Street Toronto, Ontario M5S 3E5, Canada.|Institute of Biomaterials and Biomedical Engineering, University of Toronto, 164 College Street, Toronto, Ontario M5S 3G9, Canada.
authors.forename: Alison P
authors.initials: AP
authors.lastname: McGuigan
mesh_terms: D000903:Antibiotics, Antineoplastic / Q000008:administration & dosage / Q000494:pharmacology; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects / Q000528:radiation effects; D004317:Doxorubicin / Q000008:administration & dosage / Q000494:pharmacology; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D051793:Hypoxia-Inducible Factor 1 / Q000235:genetics / Q000378:metabolism; D009369:Neoplasms / Q000378:metabolism*; D010100:Oxygen / Q000378:metabolism*; D015398:Signal Transduction; D023822:Tissue Engineering; D054457:Tissue Scaffolds
chemical_list: D000903:Antibiotics, Antineoplastic; D051793:Hypoxia-Inducible Factor 1; D004317:Doxorubicin; D010100:Oxygen