[en] Tumor organoids and patient-derived orthotopic xenografts (PDOXs) are some of the most valuable pre-clinical tools in cancer research. In this protocol, we describe efficient derivation of organoids and PDOX models from glioma patient tumors. We provide detailed steps for organoid culture, intracranial implantation, and detection of tumors in the brain. We further present technical adjustments for standardized functional assays and drug testing. For complete details on the use and execution of this protocol, please refer to Golebiewska et al. (2020).
Disciplines :
Oncologie
Auteur, co-auteur :
Oudin, Anaïs; NORLUX Neuro-Oncology Laboratory, Department of Oncology, Luxembourg Institute of Health, 1526 Luxembourg, Luxembourg.
Baus, Virginie; NORLUX Neuro-Oncology Laboratory, Department of Oncology, Luxembourg Institute of Health, 1526 Luxembourg, Luxembourg.
Barthelemy, Vanessa; NORLUX Neuro-Oncology Laboratory, Department of Oncology, Luxembourg Institute of Health, 1526 Luxembourg, Luxembourg.
Fabian, Carina; NORLUX Neuro-Oncology Laboratory, Department of Oncology, Luxembourg Institute of Health, 1526 Luxembourg, Luxembourg. ; Department of Biomedicine, University of Bergen, 5019 Bergen, Norway.
KLEIN, Eliane ; NORLUX Neuro-Oncology Laboratory, Department of Oncology, Luxembourg Institute of Health, 1526 Luxembourg, Luxembourg.
Dieterle, Monika; NORLUX Neuro-Oncology Laboratory, Department of Oncology, Luxembourg Institute of Health, 1526 Luxembourg, Luxembourg.
Wantz, May; NORLUX Neuro-Oncology Laboratory, Department of Oncology, Luxembourg Institute of Health, 1526 Luxembourg, Luxembourg.
Hau, Ann-Christin; NORLUX Neuro-Oncology Laboratory, Department of Oncology, Luxembourg Institute of Health, 1526 Luxembourg, Luxembourg.
DORDING, Claire ; NORLUX Neuro-Oncology Laboratory, Department of Oncology, Luxembourg Institute of Health, 1526 Luxembourg, Luxembourg.
Bernard, Amandine; NORLUX Neuro-Oncology Laboratory, Department of Oncology, Luxembourg Institute of Health, 1526 Luxembourg, Luxembourg. ; Neuro-Immunology Group, Department of Oncology, Luxembourg Institute of Health, 1526 Luxembourg, Luxembourg.
MICHELUCCI, Alessandro ; NORLUX Neuro-Oncology Laboratory, Department of Oncology, Luxembourg Institute of Health, 1526 Luxembourg, Luxembourg. ; Neuro-Immunology Group, Department of Oncology, Luxembourg Institute of Health, 1526 Luxembourg, Luxembourg.
Yabo, Yahaya A; NORLUX Neuro-Oncology Laboratory, Department of Oncology, Luxembourg Institute of Health, 1526 Luxembourg, Luxembourg. ; Faculty of Science, Technology and Medicine, University of Luxembourg, 4367 Belvaux, Luxembourg.
Kanli, Georgia; NORLUX Neuro-Oncology Laboratory, Department of Oncology, Luxembourg Institute of Health, 1526 Luxembourg, Luxembourg. ; Quantitative Biology Unit, Luxembourg Institute of Health, 1445 Strassen, Luxembourg. ; Translational Radiomics, Department of Oncology, Luxembourg Institute of Health, 1526 Luxembourg, Luxembourg.
Keunen, Olivier; NORLUX Neuro-Oncology Laboratory, Department of Oncology, Luxembourg Institute of Health, 1526 Luxembourg, Luxembourg. ; Quantitative Biology Unit, Luxembourg Institute of Health, 1445 Strassen, Luxembourg. ; Translational Radiomics, Department of Oncology, Luxembourg Institute of Health, 1526 Luxembourg, Luxembourg.
Bjerkvig, Rolf; NORLUX Neuro-Oncology Laboratory, Department of Oncology, Luxembourg Institute of Health, 1526 Luxembourg, Luxembourg. ; Department of Biomedicine, University of Bergen, 5019 Bergen, Norway.
NICLOU, Simone P. ; NORLUX Neuro-Oncology Laboratory, Department of Oncology, Luxembourg Institute of Health, 1526 Luxembourg, Luxembourg. ; Department of Biomedicine, University of Bergen, 5019 Bergen, Norway.
GOLEBIEWSKA, Anna ; NORLUX Neuro-Oncology Laboratory, Department of Oncology, Luxembourg Institute of Health, 1526 Luxembourg, Luxembourg.
Abdul Rahim, S.A., Dirkse, A., Oudin, A., Schuster, A., Bohler, J., Barthelemy, V., Muller, A., Vallar, L., Janji, B., Golebiewska, A., et al. Regulation of hypoxia-induced autophagy in glioblastoma involves ATG9A. Br. J. Cancer 117 (2017), 813–825, 10.1038/bjc.2017.263.
Bougnaud, S., Golebiewska, A., Oudin, A., Keunen, O., Harter, P.N., Mäder, L., Azuaje, F., Fritah, S., Stieber, D., Kaoma, T., et al. Molecular crosstalk between tumour and brain parenchyma instructs histopathological features in glioblastoma. Oncotarget 7 (2016), 31955–31971, 10.18632/oncotarget.7454.
Christensen, K., Aaberg-Jessen, C., Andersen, C., Goplen, D., Bjerkvig, R., Kristensen, B.W., Immunohistochemical expression of stem cell, endothelial cell, and chemosensitivity markers in primary glioma spheroids cultured in serum-containing and serum-free medium. Neurosurgery 66 (2010), 933–947, 10.1227/01.NEU.0000368393.45935.46.
Demeure, K., Fack, F., Duriez, E., Tiemann, K., Bernard, A., Golebiewska, A., Bougnaud, S., Bjerkvig, R., Domon, B., Niclou, S.P., Targeted proteomics to assess the response to anti-angiogenic treatment in human glioblastoma (GBM). Mol. Cell. Proteomics 15 (2016), 481–492, 10.1074/mcp.M115.052423.
Dirkse, A., Golebiewska, A., Buder, T., Nazarov, P.V., Muller, A., Poovathingal, S., Brons, N.H.C., Leite, S., Sauvageot, N., Sarkisjan, D., et al. Stem cell-associated heterogeneity in Glioblastoma results from intrinsic tumor plasticity shaped by the microenvironment. Nat. Commun., 10, 2019, 1787, 10.1038/s41467-019-09853-z.
Fack, F., Espedal, H., Keunen, O., Golebiewska, A., Obad, N., Harter, P.N., Mittelbronn, M., Bahr, O., Weyerbrock, A., Stuhr, L., et al. Bevacizumab treatment induces metabolic adaptation toward anaerobic metabolism in glioblastomas. Acta Neuropathol. 129 (2015), 115–131, 10.1007/s00401-014-1352-5.
Fack, F., Tardito, S., Hochart, G., Oudin, A., Zheng, L., Fritah, S., Golebiewska, A., Nazarov, P.V., Bernard, A., Hau, A.C., et al. Altered metabolic landscape in IDH-mutant gliomas affects phospholipid, energy, and oxidative stress pathways. EMBO Mol. Med. 9 (2017), 1681–1695, 10.15252/emmm.201707729.
Golebiewska, A., Bougnaud, S., Stieber, D., Brons, N.H., Vallar, L., Hertel, F., Klink, B., Schrock, E., Bjerkvig, R., Niclou, S.P., Side population in human glioblastoma is non-tumorigenic and characterizes brain endothelial cells. Brain 136 (2013), 1462–1475, 10.1093/brain/awt025.
Golebiewska, A., Hau, A.C., Oudin, A., Stieber, D., Yabo, Y.A., Baus, V., Barthelemy, V., Klein, E., Bougnaud, S., Keunen, O., et al. Patient-derived organoids and orthotopic xenografts of primary and recurrent gliomas represent relevant patient avatars for precision oncology. Acta Neuropathol. 140 (2020), 919–949, 10.1007/s00401-020-02226-7.
Johansson, M., Oudin, A., Tiemann, K., Bernard, A., Golebiewska, A., Keunen, O., Fack, F., Stieber, D., Wang, B., Hedman, H., Niclou, S.P., The soluble form of the tumor suppressor Lrig1 potently inhibits in vivo glioma growth irrespective of EGF receptor status. Neuro Oncol. 15 (2013), 1200–1211, 10.1093/neuonc/not054.
Keunen, O., Johansson, M., Oudin, A., Sanzey, M., Rahim, S.A., Fack, F., Thorsen, F., Taxt, T., Bartos, M., Jirik, R., et al. Anti-VEGF treatment reduces blood supply and increases tumor cell invasion in glioblastoma. Proc. Natl. Acad. Sci. U S A 108 (2011), 3749–3754, 10.1073/pnas.1014480108.
Klein, E., Hau, A.C., Oudin, A., Golebiewska, A., Niclou, S.P., Glioblastoma organoids: pre-clinical applications and challenges in the context of immunotherapy. Front. Oncol., 10, 2020, 604121, 10.3389/fonc.2020.604121.
Niclou, S.P., Danzeisen, C., Eikesdal, H.P., Wiig, H., Brons, N.H., Poli, A.M., Svendsen, A., Torsvik, A., Enger, P., Terzis, J.A., Bjerkvig, R., A novel eGFP-expressing immunodeficient mouse model to study tumor-host interactions. FASEB J. 22 (2008), 3120–3128, 10.1096/fj.08-109611.
Sanzey, M., Abdul Rahim, S.A., Oudin, A., Dirkse, A., Kaoma, T., Vallar, L., Herold-Mende, C., Bjerkvig, R., Golebiewska, A., Niclou, S.P., Comprehensive analysis of glycolytic enzymes as therapeutic targets in the treatment of glioblastoma. PLoS One, 10, 2015, e0123544, 10.1371/journal.pone.0123544.
Stieber, D., Golebiewska, A., Evers, L., Lenkiewicz, E., Brons, N.H., Nicot, N., Oudin, A., Bougnaud, S., Hertel, F., Bjerkvig, R., et al. Glioblastomas are composed of genetically divergent clones with distinct tumourigenic potential and variable stem cell-associated phenotypes. Acta Neuropathol. 127 (2014), 203–219, 10.1007/s00401-013-1196-4.
