{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:54:55Z","timestamp":1760147695845,"version":"build-2065373602"},"reference-count":25,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2023,2,17]],"date-time":"2023-02-17T00:00:00Z","timestamp":1676592000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Erich und Gertrud Roggenbuck-Stiftung","award":["R\/20.4.21"],"award-info":[{"award-number":["R\/20.4.21"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cancers"],"abstract":"<jats:p>Tumor plasticity is essential for adaptation to changing environmental conditions, in particular during the process of metastasis. In this study, we compared morphological and biochemical differences between LAN-1 neuroblastoma (NB) cells recovered from a subcutaneous xenograft primary tumor (PT) and the corresponding three generations of bone metastasis (BM I\u2013III). Moreover, growth behavior, as well as the response to chemotherapy and immune cells were assessed. For this purpose, F-actin was stained, mRNA and protein expression assessed, and lactate secretion analyzed. Further, we measured adhesion to collagen I, the growth rate of spheroids in the presence and absence of vincristine, and the production of IL-6 by peripheral blood mononuclear cells (PBMCs) co-incubated with PT or BM I\u2013III. Analysis of PT and the three BM generations revealed that their growth rate decreased from PT to BM III, and accordingly, PT cells reacted most sensitively to vincristine. In addition, morphology, adaption to hypoxic conditions, as well as transcriptomes showed strong differences between the cell lines. Moreover, BM I and BM II cells exhibited a significantly different ability to stimulate human immune cells compared to PT and BM III cells. Interestingly, the differences in immune cell stimulation corresponded to the expression level of the cancer-testis antigen MAGE-A3. In conclusion, our ex vivo model allows to analyze the adaption of tumor populations to different microenvironments and clearly demonstrates the strong alteration of tumor cell populations during this process.<\/jats:p>","DOI":"10.3390\/cancers15041274","type":"journal-article","created":{"date-parts":[[2023,2,17]],"date-time":"2023-02-17T01:32:56Z","timestamp":1676597576000},"page":"1274","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Ex Vivo Model of Neuroblastoma Plasticity"],"prefix":"10.3390","volume":"15","author":[{"given":"Paula","family":"Sch\u00e4fer","sequence":"first","affiliation":[{"name":"Department of Biochemistry and Signal Transduction, University Medical Center Hamburg-Eppendorf, Martinistrasse 52, D-20246 Hamburg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Stefanie","family":"Muhs","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Signal Transduction, University Medical Center Hamburg-Eppendorf, Martinistrasse 52, D-20246 Hamburg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lucas","family":"Turnbull","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Signal Transduction, University Medical Center Hamburg-Eppendorf, Martinistrasse 52, D-20246 Hamburg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Palwasha","family":"Garwal","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Signal Transduction, University Medical Center Hamburg-Eppendorf, Martinistrasse 52, D-20246 Hamburg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hanna","family":"Maar","sequence":"additional","affiliation":[{"name":"Institute of Anatomy and Experimental Morphology, University Medical Center Hamburg-Eppendorf, Martinistrasse 52, D-20246 Hamburg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0712-0983","authenticated-orcid":false,"given":"Timur A.","family":"Yorgan","sequence":"additional","affiliation":[{"name":"Department of Osteology and Biomechanics, University Medical Center Hamburg-Eppendorf, Martinistrasse 52, D-20246 Hamburg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eva","family":"Tolosa","sequence":"additional","affiliation":[{"name":"Department of Immunology, University Medical Center Hamburg-Eppendorf, Martinistrasse 52, D-20246 Hamburg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tobias","family":"Lange","sequence":"additional","affiliation":[{"name":"Institute of Anatomy and Experimental Morphology, University Medical Center Hamburg-Eppendorf, Martinistrasse 52, D-20246 Hamburg, Germany"},{"name":"Institute of Anatomy I, Jena University Hospital, Teichgraben 7, D-07743 Jena, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sabine","family":"Windhorst","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Signal Transduction, University Medical Center Hamburg-Eppendorf, Martinistrasse 52, D-20246 Hamburg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2023,2,17]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Van Arendonk, K.J., and Chung, D.H. (2019). Neuroblastoma: Tumor Biology and Its Implications for Staging and Treatment. Children, 6.","DOI":"10.3390\/children6010012"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"11234","DOI":"10.1158\/0008-5472.CAN-07-0718","article-title":"Neuroblastoma cells isolated from bone marrow metastases contain a naturally enriched tumor-initiating cell","volume":"67","author":"Hansford","year":"2007","journal-title":"Cancer Res."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"419","DOI":"10.1016\/j.scr.2015.08.008","article-title":"A distinct gene expression signature characterizes human neuroblastoma cancer stem cells","volume":"15","author":"Ross","year":"2015","journal-title":"Stem Cell Res."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"838","DOI":"10.1593\/neo.04310","article-title":"Characteristics of stem cells from human neuroblastoma cell lines and in tumors","volume":"6","author":"Walton","year":"2004","journal-title":"Neoplasia"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1408","DOI":"10.1038\/ng.3921","article-title":"Heterogeneity of neuroblastoma cell identity defined by transcriptional circuitries","volume":"49","author":"Boeva","year":"2017","journal-title":"Nat. Genet."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1261","DOI":"10.1038\/ng.3899","article-title":"Neuroblastoma is composed of two super-enhancer-associated differentiation states","volume":"49","author":"Koster","year":"2017","journal-title":"Nat. Genet."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Gautier, M., Thirant, C., Delattre, O., and Janoueix-Lerosey, I. (2021). Plasticity in Neuroblastoma Cell Identity Defines a Noradrenergic-to-Mesenchymal Transition (NMT). Cancers, 13.","DOI":"10.3390\/cancers13122904"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1038\/s41573-019-0044-1","article-title":"The great escape: Tumour cell plasticity in resistance to targeted therapy","volume":"19","author":"Boumahdi","year":"2020","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Labitzky, V., Baranowsky, A., Maar, H., Hanika, S., Starzonek, S., Ahlers, A.K., St\u00fcbke, K., Koziolek, E.J., Heine, M., and Sch\u00e4fer, P. (2020). Modeling Spontaneous Bone Metastasis Formation of Solid Human Tumor Xenografts in Mice. Cancers, 12.","DOI":"10.3390\/cancers12020385"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"5166","DOI":"10.1091\/mbc.e09-01-0083","article-title":"Neuronal IP3 3-kinase is an F-actin-bundling protein: Role in dendritic targeting and regulation of spine morphology","volume":"20","author":"Johnson","year":"2009","journal-title":"Mol. Biol. Cell"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"269","DOI":"10.1038\/nature13034","article-title":"Direct measurement of local oxygen concentration in the bone marrow of live animals","volume":"508","author":"Spencer","year":"2014","journal-title":"Nature"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"114700","DOI":"10.1016\/j.bcp.2021.114700","article-title":"Oncogenic activity and cellular functionality of melanoma associated antigen A3","volume":"192","author":"Schafer","year":"2021","journal-title":"Biochem. Pharmacol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"963","DOI":"10.1016\/j.molcel.2010.08.029","article-title":"MAGE-RING protein complexes comprise a family of E3 ubiquitin ligases","volume":"39","author":"Doyle","year":"2010","journal-title":"Mol. Cell"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"844","DOI":"10.1080\/15548627.2015.1034420","article-title":"Oncogenic MAGEA-TRIM28 ubiquitin ligase downregulates autophagy by ubiquitinating and degrading AMPK in cancer","volume":"11","author":"Pineda","year":"2015","journal-title":"Autophagy"},{"key":"ref_15","first-page":"127","article-title":"Expression of p53 in human neuroblastoma- and neuroepithelioma-derived cell lines","volume":"7","author":"Davidoff","year":"1992","journal-title":"Oncogene"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"390","DOI":"10.1002\/ijc.2910390320","article-title":"Production of stable cytolytic T-cell clones directed against autologous human melanoma","volume":"39","author":"Herin","year":"1987","journal-title":"Int. J. Cancer"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1702","DOI":"10.7150\/ijms.27643","article-title":"Cancer\/testis Antigen MAGEA3 Interacts with STAT1 and Remodels the Tumor Microenvironment","volume":"15","author":"Wang","year":"2018","journal-title":"Int. J. Med. Sci."},{"key":"ref_18","unstructured":"Tolosa, E. Personal observation."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1895","DOI":"10.1056\/NEJMp1508811","article-title":"The Path to Cancer\u2014Three Strikes and You\u2019re Out","volume":"373","author":"Vogelstein","year":"2015","journal-title":"N. Engl. J. Med."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"569","DOI":"10.1038\/s12276-020-0423-z","article-title":"The force awakens: Metastatic dormant cancer cells","volume":"52","author":"Park","year":"2020","journal-title":"Exp. Mol. Med."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"3840","DOI":"10.1158\/1078-0432.CCR-06-2507","article-title":"Detection of MAGE-A transcripts in bone marrow is an independent prognostic factor in operable non-small-cell lung cancer","volume":"13","author":"Sienel","year":"2007","journal-title":"Clin. Cancer Res."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Wright, S.C.E., Vasilevski, N., Serra, V., Rodon, J., and Eichhorn, P.J.A. (2021). Mechanisms of Resistance to PI3K Inhibitors in Cancer: Adaptive Responses, Drug Tolerance and Cellular Plasticity. Cancers, 13.","DOI":"10.3390\/cancers13071538"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"aaa6566","DOI":"10.1126\/science.aaa6566","article-title":"Innate lymphoid cells. Innate lymphoid cells: A new paradigm in immunology","volume":"348","author":"Eberl","year":"2015","journal-title":"Science"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"916","DOI":"10.1016\/S1470-2045(18)30254-7","article-title":"MAGE-A3 immunotherapeutic as adjuvant therapy for patients with resected, MAGE-A3-positive, stage III melanoma (DERMA): A double-blind, randomised, placebo-controlled, phase 3 trial","volume":"19","author":"Dreno","year":"2018","journal-title":"Lancet Oncol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"822","DOI":"10.1016\/S1470-2045(16)00099-1","article-title":"Efficacy of the MAGE-A3 cancer immunotherapeutic as adjuvant therapy in patients with resected MAGE-A3-positive non-small-cell lung cancer (MAGRIT): A randomised, double-blind, placebo-controlled, phase 3 trial","volume":"17","author":"Vansteenkiste","year":"2016","journal-title":"Lancet Oncol."}],"container-title":["Cancers"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6694\/15\/4\/1274\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T18:38:50Z","timestamp":1760121530000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6694\/15\/4\/1274"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,2,17]]},"references-count":25,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2023,2]]}},"alternative-id":["cancers15041274"],"URL":"https:\/\/doi.org\/10.3390\/cancers15041274","relation":{},"ISSN":["2072-6694"],"issn-type":[{"type":"electronic","value":"2072-6694"}],"subject":[],"published":{"date-parts":[[2023,2,17]]}}}