{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,16]],"date-time":"2025-10-16T19:50:07Z","timestamp":1760644207328,"version":"build-2065373602"},"reference-count":52,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2024,1,19]],"date-time":"2024-01-19T00:00:00Z","timestamp":1705622400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Friends of The Royal Melbourne Hospital Neuroscience Foundation"},{"name":"Australian Federal Government"},{"name":"Australian Rotary Health and Dine for a Cure"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Background: Glioblastoma is characterised by extensive infiltration into the brain parenchyma, leading to inevitable tumor recurrence and therapeutic failure. Future treatments will need to target the specific biology of tumour recurrence, but our current understanding of the underlying mechanisms is limited. Significantly, there is a lack of available methods and models that are tailored to the examination of tumour recurrence. Methods: NOD-SCID mice were orthotopically implanted with luciferase-labelled donor U87MG or MU20 glioblastoma cells. Four days later, an unlabelled recipient tumor was implanted on the contralateral side. The mice were euthanised at a humane end-point and tissue and blood samples were collected for ex vivo analyses. Results: The ex vivo analyses of the firefly-labelled MU20 tumours displayed extensive invasion at the primary tumour margins, whereas the firefly-labelled U87MG tumours exhibited expansive phenotypes with no evident invasions at the tumour margins. Luciferase signals were detected in the contralateral unlabelled recipient tumours for both the U87MG and MU20 tumours compared to the non-implanted control brain. Remarkably, tumour cells were uniformly detected in all tissue samples of the supratentorial brain region compared to the control tissue, with single tumour cells detected in some tissue samples. Circulating tumour cells were also detected in the blood samples of most of the xenografted mice. Moreover, tumour cells were detected in the lungs of all of the mice, a probable event related to haematogenous dissemination. Similar results were obtained when the U87MG cells were alternatively labelled with gaussian luciferase. Conclusions: These findings describe a systemic disease model for glioblastoma which can be used to investigate recurrence biology and therapeutic efficacy towards recurrence.<\/jats:p>","DOI":"10.3390\/cells13020192","type":"journal-article","created":{"date-parts":[[2024,1,19]],"date-time":"2024-01-19T07:00:32Z","timestamp":1705647632000},"page":"192","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["A New Systemic Disease Mouse Model for Glioblastoma Capable of Single-Tumour-Cell Detection"],"prefix":"10.3390","volume":"13","author":[{"given":"Thomas M. B.","family":"Ware","sequence":"first","affiliation":[{"name":"Department of Surgery, The Royal Melbourne Hospital, The University of Melbourne, Parkville, VIC 3050, Australia"},{"name":"Huagene Institute, Kecheng Science and Technology Park, Pukou District, Nanjing 211806, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3020-4245","authenticated-orcid":false,"given":"Rodney B.","family":"Luwor","sequence":"additional","affiliation":[{"name":"Department of Surgery, The Royal Melbourne Hospital, The University of Melbourne, Parkville, VIC 3050, Australia"},{"name":"Huagene Institute, Kecheng Science and Technology Park, Pukou District, Nanjing 211806, China"},{"name":"Fiona Elsey Cancer Research Institute, Ballarat, VIC 3350, Australia"},{"name":"Federation University, Ballarat, VIC 3350, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hong-Jian","family":"Zhu","sequence":"additional","affiliation":[{"name":"Department of Surgery, The Royal Melbourne Hospital, The University of Melbourne, Parkville, VIC 3050, Australia"},{"name":"Huagene Institute, Kecheng Science and Technology Park, Pukou District, Nanjing 211806, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,1,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"iv1","DOI":"10.1093\/neuonc\/noy131","article-title":"CBTRUS Statistical Report: Primary Brain and Other Central Nervous System Tumors Diagnosed in the United States in 2011\u20132015","volume":"20","author":"Ostrom","year":"2018","journal-title":"Neuro Oncol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"459","DOI":"10.1016\/S1470-2045(09)70025-7","article-title":"Effects of radiotherapy with concomitant and adjuvant temozolomide versus radiotherapy alone on survival in glioblastoma in a randomised phase III study: 5-year analysis of the EORTC-NCIC trial","volume":"10","author":"Stupp","year":"2009","journal-title":"Lancet Oncol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1193","DOI":"10.1016\/j.drudis.2019.03.008","article-title":"Therapy for glioblastoma: Is it working?","volume":"24","author":"Zanders","year":"2019","journal-title":"Drug Discov. Today"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1038\/nbt.2786","article-title":"Clinical development success rates for investigational drugs","volume":"32","author":"Hay","year":"2014","journal-title":"Nat. Biotechnol."},{"key":"ref_5","first-page":"1","article-title":"Glioblastoma treatment: Where to now?","volume":"5","author":"Ware","year":"2018","journal-title":"Integr. Cancer Sci. Ther."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"6095","DOI":"10.1038\/sj.onc.1207870","article-title":"The tumor-specific de2-7 epidermal growth factor receptor (EGFR) promotes cells survival and heterodimerizes with the wild-type EGFR","volume":"23","author":"Luwor","year":"2004","journal-title":"Oncogene"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"98","DOI":"10.1016\/j.ccr.2009.12.020","article-title":"Integrated genomic analysis identifies clinically relevant subtypes of glioblastoma characterized by abnormalities in PDGFRA, IDH1, EGFR, and NF1","volume":"17","author":"Verhaak","year":"2010","journal-title":"Cancer Cell"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"2264","DOI":"10.1158\/1078-0432.CCR-05-2510","article-title":"Use of an orthotopic xenograft model for assessing the effect of epidermal growth factor receptor amplification on glioblastoma radiation response","volume":"12","author":"Sarkaria","year":"2006","journal-title":"Clin. Cancer Res."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1227\/01.NEU.0000145865.25689.55","article-title":"Anti-epidermal growth factor receptor monoclonal antibody cetuximab augments radiation effects in glioblastoma multiforme in vitro and in vivo","volume":"56","author":"Eller","year":"2005","journal-title":"Neurosurgery"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1268","DOI":"10.1200\/JCO.2008.17.5984","article-title":"Randomized phase II trial of erlotinib versus temozolomide or carmustine in recurrent glioblastoma: EORTC brain tumor group study 26034","volume":"27","author":"Brandes","year":"2009","journal-title":"J. Clin. Oncol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1093\/neuonc\/nop015","article-title":"A phase II trial of erlotinib in patients with recurrent malignant gliomas and nonprogressive glioblastoma multiforme postradiation therapy","volume":"12","author":"Raizer","year":"2010","journal-title":"Neuro Oncol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1200\/JCO.2004.08.110","article-title":"Phase II trial of gefitinib in recurrent glioblastoma","volume":"22","author":"Rich","year":"2004","journal-title":"J. Clin. Oncol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1304","DOI":"10.1227\/01.NEU.0000245622.70344.BE","article-title":"Effect of the angiogenesis inhibitor Cilengitide (EMD 121974) on glioblastoma growth in nude mice","volume":"59","author":"Yamada","year":"2006","journal-title":"Neurosurgery"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"5610","DOI":"10.1200\/JCO.2008.16.7510","article-title":"Randomized phase II study of cilengitide, an integrin-targeting arginine-glycine-aspartic acid peptide, in recurrent glioblastoma multiforme","volume":"26","author":"Reardon","year":"2008","journal-title":"J. Clin. Oncol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"708","DOI":"10.1093\/neuonc\/nou356","article-title":"Two cilengitide regimens in combination with standard treatment for patients with newly diagnosed glioblastoma and unmethylated MGMT gene promoter: Results of the open-label, controlled, randomized phase II CORE study","volume":"17","author":"Nabors","year":"2015","journal-title":"Neuro Oncol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1100","DOI":"10.1016\/S1470-2045(14)70379-1","article-title":"Cilengitide combined with standard treatment for patients with newly diagnosed glioblastoma with methylated MGMT promoter (CENTRIC EORTC 26071-22072 study): A multicentre, randomised, open-label, phase 3 trial","volume":"15","author":"Stupp","year":"2014","journal-title":"Lancet Oncol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"7843","DOI":"10.1158\/0008-5472.CAN-06-1010","article-title":"Stem cell-like glioma cells promote tumor angiogenesis through vascular endothelial growth factor","volume":"66","author":"Bao","year":"2006","journal-title":"Cancer Res."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1383","DOI":"10.1593\/neo.08928","article-title":"Combining bevacizumab with temozolomide increases the antitumor efficacy of temozolomide in a human glioblastoma orthotopic xenograft model","volume":"10","author":"Mathieu","year":"2008","journal-title":"Neoplasia"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"4733","DOI":"10.1200\/JCO.2008.19.8721","article-title":"Bevacizumab alone and in combination with irinotecan in recurrent glioblastoma","volume":"27","author":"Friedman","year":"2009","journal-title":"J. Clin. Oncol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"699","DOI":"10.1056\/NEJMoa1308573","article-title":"A randomized trial of bevacizumab for newly diagnosed glioblastoma","volume":"370","author":"Gilbert","year":"2014","journal-title":"N. Engl. J. Med."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"709","DOI":"10.1056\/NEJMoa1308345","article-title":"Bevacizumab plus radiotherapy-temozolomide for newly diagnosed glioblastoma","volume":"370","author":"Chinot","year":"2014","journal-title":"N. Engl. J. Med."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1611","DOI":"10.1200\/JCO.2015.63.4691","article-title":"Bevacizumab Plus Irinotecan Versus Temozolomide in Newly Diagnosed O6-Methylguanine-DNA Methyltransferase Nonmethylated Glioblastoma: The Randomized GLARIUS Trial","volume":"34","author":"Herrlinger","year":"2016","journal-title":"J. Clin. Oncol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1093\/neuonc\/nop027","article-title":"Tumor invasion after treatment of glioblastoma with bevacizumab: Radiographic and pathologic correlation in humans and mice","volume":"12","author":"Fuller","year":"2010","journal-title":"Neuro Oncol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"264","DOI":"10.1002\/ana.22036","article-title":"Residual tumor cells are unique cellular targets in glioblastoma","volume":"68","author":"Glas","year":"2010","journal-title":"Ann. Neurol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"5819","DOI":"10.1038\/onc.2016.85","article-title":"A comprehensive profile of recurrent glioblastoma","volume":"35","author":"Campos","year":"2016","journal-title":"Oncogene"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"2054","DOI":"10.1038\/s41598-019-38625-4","article-title":"Intravital imaging of glioma border morphology reveals distinctive cellular dynamics and contribution to tumor cell invasion","volume":"9","author":"Alieva","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1306","DOI":"10.1002\/glia.20850","article-title":"Imaging glioma cell invasion in vivo reveals mechanisms of dissemination and peritumoral angiogenesis","volume":"57","author":"Winkler","year":"2009","journal-title":"Glia"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"491","DOI":"10.1038\/sj.neo.7900121","article-title":"Rapid and quantitative assessment of cancer treatment response using in vivo bioluminescence imaging","volume":"2","author":"Rehemtulla","year":"2000","journal-title":"Neoplasia"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"243","DOI":"10.1016\/j.canlet.2004.07.010","article-title":"Monitoring luciferase-labeled cancer cell growth and metastasis in different in vivo models","volume":"217","author":"Nogawa","year":"2005","journal-title":"Cancer Lett."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"11003","DOI":"10.1038\/s41598-020-67411-w","article-title":"GL261 luciferase-expressing cells elicit an anti-tumor immune response: An evaluation of murine glioma models","volume":"10","author":"Sanchez","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"161","DOI":"10.2310\/7290.2012.00029","article-title":"In vivo bioluminescence imaging validation of a human biopsy-derived orthotopic mouse model of glioblastoma multiforme","volume":"12","author":"Jarzabek","year":"2013","journal-title":"Mol. Imaging"},{"key":"ref_32","first-page":"e53287","article-title":"Bioluminescence Imaging of an Immunocompetent Animal Model for Glioblastoma","volume":"107","author":"Clark","year":"2016","journal-title":"J. Vis. Exp."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1791","DOI":"10.7150\/jca.15564","article-title":"Noninvasive Monitoring of Glioma Growth in the Mouse","volume":"7","author":"Alessandrini","year":"2016","journal-title":"J. Cancer"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Sasportas, L.S., Hori, S.S., Pratx, G., and Gambhir, S.S. (2014). Detection and quantitation of circulating tumor cell dynamics by bioluminescence imaging in an orthotopic mammary carcinoma model. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0105079"},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Shuryak, I., Smilenov, L.B., Kleiman, N.J., and Brenner, D.J. (2013). Potential reduction of contralateral second breast-cancer risks by prophylactic mammary irradiation: Validation in a breast-cancer-prone mouse model. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0085795"},{"key":"ref_36","first-page":"e975572","article-title":"Cross-talk between tumors can affect responses to therapy","volume":"4","author":"Devaud","year":"2015","journal-title":"Oncoimmunology"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"456","DOI":"10.1038\/s41392-023-01711-1","article-title":"Simultaneously targeting extracellular vesicle trafficking and TGF-beta receptor kinase activity blocks signaling hyperactivation and metastasis","volume":"8","author":"Teixeira","year":"2023","journal-title":"Signal Transduct. Target Ther."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"2152","DOI":"10.1158\/0008-5472.CAN-13-0813","article-title":"Detection of brain tumor cells in the peripheral blood by a telomerase promoter-based assay","volume":"74","author":"Macarthur","year":"2014","journal-title":"Cancer Res."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1299","DOI":"10.1158\/2159-8290.CD-14-0471","article-title":"Brain tumor cells in circulation are enriched for mesenchymal gene expression","volume":"4","author":"Sullivan","year":"2014","journal-title":"Cancer Discov."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"823","DOI":"10.1001\/jama.1928.02690380007003","article-title":"Removal of Right Cerebral Hemisphere for Certain Tumors with Hemiplegia: Preliminary Report","volume":"90","author":"Dandy","year":"1928","journal-title":"J. Am. Med. Assoc."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"4","DOI":"10.1093\/neuonc\/nos273","article-title":"Standards of care for treatment of recurrent glioblastoma\u2014Are we there yet?","volume":"15","author":"Weller","year":"2013","journal-title":"Neuro Oncol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"523","DOI":"10.1001\/archneurol.2011.2910","article-title":"Addressing Diffuse Glioma as a Systemic Brain Disease With Single-Cell Analysis","volume":"69","author":"Sahm","year":"2012","journal-title":"Arch. Neurol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1807","DOI":"10.1126\/science.1164382","article-title":"An integrated genomic analysis of human glioblastoma multiforme","volume":"321","author":"Parsons","year":"2008","journal-title":"Science"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"765","DOI":"10.1056\/NEJMoa0808710","article-title":"IDH1 and IDH2 mutations in gliomas","volume":"360","author":"Yan","year":"2009","journal-title":"N. Engl. J. Med."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"979","DOI":"10.1093\/neuonc\/nos135","article-title":"In vivo models of primary brain tumors: Pitfalls and perspectives","volume":"14","author":"Huszthy","year":"2012","journal-title":"Neuro Oncol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"714","DOI":"10.1016\/j.ctrv.2009.08.011","article-title":"High-grade glioma mouse models and their applicability for preclinical testing","volume":"35","author":"Beijnen","year":"2009","journal-title":"Cancer Treat. Rev."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1136\/mp.56.3.187","article-title":"No evidence of tumour cells in blood of patients with glioma","volume":"56","author":"Bohm","year":"2003","journal-title":"J. Clin. Pathol. Mol. Pathol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"124","DOI":"10.1227\/00006123-199501000-00016","article-title":"Glioblastoma cells do not intravasate into blood vessels","volume":"36","author":"Bernstein","year":"1995","journal-title":"Neurosurgery"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"247ra101","DOI":"10.1126\/scitranslmed.3009095","article-title":"Hematogenous dissemination of glioblastoma multiforme","volume":"6","author":"Muller","year":"2014","journal-title":"Sci. Transl. Med."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"487","DOI":"10.1038\/s41416-018-0186-7","article-title":"Detection of circulating tumour cell clusters in human glioblastoma","volume":"119","author":"Krol","year":"2018","journal-title":"Br. J. Cancer"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"6632","DOI":"10.1158\/0008-5472.CAN-18-0650","article-title":"Circulating Glioma Cells Exhibit Stem Cell-like Properties","volume":"78","author":"Liu","year":"2018","journal-title":"Cancer Res."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"692","DOI":"10.1016\/j.ccell.2019.02.007","article-title":"Evolutionary Trajectories of IDHWT Glioblastomas Reveal a Common Path of Early Tumorigenesis Instigated Years ahead of Initial Diagnosis","volume":"35","author":"Yang","year":"2019","journal-title":"Cancer Cell"}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/13\/2\/192\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T13:46:01Z","timestamp":1760103961000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/13\/2\/192"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,1,19]]},"references-count":52,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2024,1]]}},"alternative-id":["cells13020192"],"URL":"https:\/\/doi.org\/10.3390\/cells13020192","relation":{},"ISSN":["2073-4409"],"issn-type":[{"type":"electronic","value":"2073-4409"}],"subject":[],"published":{"date-parts":[[2024,1,19]]}}}