title: A 3D model of tumour angiogenic microenvironment to monitor hypoxia effects on cell interactions and cancer stem cell selection.
abstract: Tumour microenvironment determines the fate of treatments. Reconstitution of tumour conditions is mandatory for alternative in vitro methods devoted to cancer development and the selection of therapeutic strategies. This work describes a 3D model of melanoma growth in its environment. Introducing means to mimic tumour angiogenesis, which turns on tumour progression, the model shows that melanoma tumour spheroids allow reconstitution of solid tumours with stromal cells. Angiogenesis evidenced the differential recruitment of endothelial cells (EC) from early progenitors (EEPCs) to mature ECs. Hypoxia was the key parameter that selected and stabilized melanoma cancer stem like cells (CSCs) phenotype based on aldehyde dehydrogenase expression as the best criterion. The 3D-tumour-model demonstrated the distinct reactivity of ECs toward tumour cells in terms of cellular cross-talk and humoral response. Intra-spheroid cell-to-cell membrane dye exchanges, mediated by intercellular interactions, uncovered the melanoma-to-EEPC cooperation. The resulting changes in tumour milieu were evidenced by the chemokinic composition and hypoxia-related variations in microRNA expression assessed in each cellular component of the spheroids. This method brings new tools to decipher the molecular mechanism of tumour-mediated cell recruitment and for in vitro assessment of therapeutic approaches.
authors.affiliation: Centre for Molecular Biophysics, UPR 4301 CNRS Affiliated to Orléans University and INSERM, Orléans Cedex 02, France; Department of Biophysics, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, ul. Gronostajowa 7, 30387, Kraków, Poland.
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Klimkiewicz
authors.affiliation: Centre for Molecular Biophysics, UPR 4301 CNRS Affiliated to Orléans University and INSERM, Orléans Cedex 02, France.
authors.forename: Kazimierz
authors.initials: K
authors.lastname: Weglarczyk
authors.affiliation: Centre for Molecular Biophysics, UPR 4301 CNRS Affiliated to Orléans University and INSERM, Orléans Cedex 02, France.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Collet
authors.affiliation: Laboratory of Glycobiology and Cell Recognition, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, PAN, U. Rudolfa Weigla 12, 53114, Wrocław, Poland.
authors.forename: Maria
authors.initials: M
authors.lastname: Paprocka
authors.affiliation: Centre for Molecular Biophysics, UPR 4301 CNRS Affiliated to Orléans University and INSERM, Orléans Cedex 02, France.
authors.forename: Alan
authors.initials: A
authors.lastname: Guichard
authors.affiliation: Department of Biophysics, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, ul. Gronostajowa 7, 30387, Kraków, Poland.
authors.forename: Michal
authors.initials: M
authors.lastname: Sarna
authors.affiliation: Department of Medical Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, ul. Gronostajowa 7, 30387, Kraków, Poland.
authors.forename: Alicja
authors.initials: A
authors.lastname: Jozkowicz
authors.affiliation: Department of Medical Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, ul. Gronostajowa 7, 30387, Kraków, Poland; Malopolska Centre of Biotechnology, Jagiellonian University, ul. Gronostajowa 7A, 30-387, Krakow, Poland.
authors.forename: Jozef
authors.initials: J
authors.lastname: Dulak
authors.affiliation: Department of Biophysics, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, ul. Gronostajowa 7, 30387, Kraków, Poland.
authors.forename: Tadeusz
authors.initials: T
authors.lastname: Sarna
authors.affiliation: Centre for Molecular Biophysics, UPR 4301 CNRS Affiliated to Orléans University and INSERM, Orléans Cedex 02, France.
authors.forename: Catherine
authors.initials: C
authors.lastname: Grillon
authors.affiliation: Centre for Molecular Biophysics, UPR 4301 CNRS Affiliated to Orléans University and INSERM, Orléans Cedex 02, France; Malopolska Centre of Biotechnology, Jagiellonian University, ul. Gronostajowa 7A, 30-387, Krakow, Poland. Electronic address: claudine.kieda@cnrs-orleans.fr.
authors.forename: Claudine
authors.initials: C
authors.lastname: Kieda
mesh_terms: D000818:Animals; D002450:Cell Communication / Q000502:physiology*; D015687:Cell Hypoxia / Q000502:physiology*; D049109:Cell Proliferation / Q000502:physiology; D006801:Humans; D021621:Imaging, Three-Dimensional; D008545:Melanoma / Q000098:blood supply* / Q000378:metabolism / Q000473:pathology*; D008546:Melanoma, Experimental / Q000098:blood supply / Q000378:metabolism / Q000473:pathology; D051379:Mice; D014411:Neoplastic Stem Cells / Q000378:metabolism* / Q000473:pathology*; D009389:Neovascularization, Pathologic / Q000378:metabolism / Q000473:pathology; D018874:Spheroids, Cellular; D059016:Tumor Microenvironment
keywords: Angiogenesis; Endothelial progenitors; Hypoxia; Recruitment; Spheroid tumour model; microRNA