{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T12:06:37Z","timestamp":1789733197239,"version":"4.0.1"},"reference-count":61,"publisher":"Wiley","license":[{"start":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T00:00:00Z","timestamp":1789689600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T00:00:00Z","timestamp":1789689600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["FL 865\/2\u20101"],"award-info":[{"award-number":["FL 865\/2\u20101"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["febs.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Molecular Oncology"],"abstract":"<jats:p>\n                    Circulating tumor cells (CTCs) are clinically relevant markers of metastasis and prognosis in gastrointestinal (GI) cancers. However, their extreme rarity and the limitations of current\n                    <jats:italic>in vivo<\/jats:italic>\n                    models severely restrict mechanistic studies of tumor cell dissemination. Here, we demonstrate for the first time that the avian chorioallantoic membrane (CAM) assay can serve as a biologically relevant and ethically advantageous 3R\u2010compliant (Replacement, Reduction, and Refinement)\n                    <jats:italic>in vivo<\/jats:italic>\n                    model for investigating spontaneous CTC generation. Six green fluorescent protein (GFP)\u2010labeled human colorectal, pancreatic, and cholangiocarcinoma cell lines were xenografted onto the CAM. Using a standardized workflow, we achieved reliable detection and isolation of GFP\u2010positive single CTCs from small\u2010volume CAM blood samples. Critically, we reproducibly confirmed the presence of spontaneously shed CAM\u2010derived CTCs independent of GFP expression using three established, clinically relevant CTC detection platforms. Subsequent single\u2010cell whole\u2010genome amplification enabled downstream short\u2010tandem repeat (STR) profiling to authenticate CTC identity, alongside detection of tumor\u2010specific point mutations and copy number alterations (CNAs). Isolated CTCs exhibited high genetic concordance with their parental cell lines, validating the technical robustness of our workflow and establishing the CAM assay as a versatile, scalable platform for investigating hematogenous dissemination and CTC biology outside of mammalian models. Consequently, the CAM assay opens new avenues for early mechanistic and pharmacological CTC research, including functional studies of patient\u2010derived material.\n                  <\/jats:p>","DOI":"10.1002\/1878-0261.70336","type":"journal-article","created":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T11:50:37Z","timestamp":1789732237000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Bridging the gap: a genetically validated avian chorioallantoic membrane platform for investigation of spontaneous circulating tumor cells"],"prefix":"10.1002","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7044-8543","authenticated-orcid":false,"given":"Dennis","family":"Roth","sequence":"first","affiliation":[{"name":"Department of Surgery (A) Heinrich\u2010Heine\u2010University and University Hospital Duesseldorf  Germany"},{"name":"Liquid Biopsy Center Duesseldorf Center for Integrated Oncology Aachen  Bonn, Cologne Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4515-739X","authenticated-orcid":false,"given":"Rui P. L.","family":"Neves","sequence":"additional","affiliation":[{"name":"Department of Surgery (A) Heinrich\u2010Heine\u2010University and University Hospital Duesseldorf  Germany"},{"name":"Liquid Biopsy Center Duesseldorf Center for Integrated Oncology Aachen  Bonn, Cologne Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julia","family":"Geheb","sequence":"additional","affiliation":[{"name":"Department of Surgery (A) Heinrich\u2010Heine\u2010University and University Hospital Duesseldorf  Germany"},{"name":"Liquid Biopsy Center Duesseldorf Center for Integrated Oncology Aachen  Bonn, Cologne Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christian","family":"Ballmeyer","sequence":"additional","affiliation":[{"name":"Department of Surgery (A) Heinrich\u2010Heine\u2010University and University Hospital Duesseldorf  Germany"},{"name":"Liquid Biopsy Center Duesseldorf Center for Integrated Oncology Aachen  Bonn, Cologne Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nathalie","family":"Ga\u00dfmann","sequence":"additional","affiliation":[{"name":"Department of Surgery (A) Heinrich\u2010Heine\u2010University and University Hospital Duesseldorf  Germany"},{"name":"Liquid Biopsy Center Duesseldorf Center for Integrated Oncology Aachen  Bonn, Cologne Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kiki C.","family":"Andree","sequence":"additional","affiliation":[{"name":"VyCAP B.V  Enschede The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Monica","family":"Sudarsanam","sequence":"additional","affiliation":[{"name":"Department of Surgery (A) Heinrich\u2010Heine\u2010University and University Hospital Duesseldorf  Germany"},{"name":"Liquid Biopsy Center Duesseldorf Center for Integrated Oncology Aachen  Bonn, Cologne Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christiane","family":"Driemel","sequence":"additional","affiliation":[{"name":"Department of Surgery (A) Heinrich\u2010Heine\u2010University and University Hospital Duesseldorf  Germany"},{"name":"Liquid Biopsy Center Duesseldorf Center for Integrated Oncology Aachen  Bonn, Cologne Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Celine","family":"\u00d6rencik","sequence":"additional","affiliation":[{"name":"Department of Surgery (A) Heinrich\u2010Heine\u2010University and University Hospital Duesseldorf  Germany"},{"name":"Liquid Biopsy Center Duesseldorf Center for Integrated Oncology Aachen  Bonn, Cologne Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andr\u00e8","family":"Franken","sequence":"additional","affiliation":[{"name":"Liquid Biopsy Center Duesseldorf Center for Integrated Oncology Aachen  Bonn, Cologne Germany"},{"name":"Department of Obstetrics and Gynecology University Hospital and Medical Faculty of the Heinrich\u2010Heine\u2010University Duesseldorf  Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eslam","family":"Elmaghraby","sequence":"additional","affiliation":[{"name":"Department of Surgery (A) Heinrich\u2010Heine\u2010University and University Hospital Duesseldorf  Germany"},{"name":"Liquid Biopsy Center Duesseldorf Center for Integrated Oncology Aachen  Bonn, Cologne Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sven H.","family":"Loosen","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Hepatology and Infectious Diseases University Hospital Duesseldorf, Medical Faculty of Heinrich Heine University Duesseldorf  Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Roderburg","family":"Christoph","sequence":"additional","affiliation":[{"name":"Department of Gastroenterology, Hepatology and Infectious Diseases University Hospital Duesseldorf, Medical Faculty of Heinrich Heine University Duesseldorf  Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sebastian","family":"Schoelch","sequence":"additional","affiliation":[{"name":"JCCU Translational Surgical Oncology (A430) German Cancer Research Center (DKFZ)  Heidelberg Germany"},{"name":"DKFZ\u2010Hector Cancer Institute, Medical Faculty Mannheim Heidelberg University  Mannheim Germany"},{"name":"Department of Surgery Universit\u00e4tsmedizin Mannheim, Medical Faculty Mannheim, Heidelberg University  Mannheim"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hans","family":"Neubauer","sequence":"additional","affiliation":[{"name":"Liquid Biopsy Center Duesseldorf Center for Integrated Oncology Aachen  Bonn, Cologne Germany"},{"name":"Department of Obstetrics and Gynecology University Hospital and Medical Faculty of the Heinrich\u2010Heine\u2010University Duesseldorf  Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wolfram T.","family":"Knoefel","sequence":"additional","affiliation":[{"name":"Department of Surgery (A) Heinrich\u2010Heine\u2010University and University Hospital Duesseldorf  Germany"},{"name":"Liquid Biopsy Center Duesseldorf Center for Integrated Oncology Aachen  Bonn, Cologne Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3412-594X","authenticated-orcid":false,"given":"Nikolas H.","family":"Stoecklein","sequence":"additional","affiliation":[{"name":"Department of Surgery (A) Heinrich\u2010Heine\u2010University and University Hospital Duesseldorf  Germany"},{"name":"Liquid Biopsy Center Duesseldorf Center for Integrated Oncology Aachen  Bonn, Cologne Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9322-1659","authenticated-orcid":false,"given":"Georg","family":"Fluegen","sequence":"additional","affiliation":[{"name":"Department of Surgery (A) Heinrich\u2010Heine\u2010University and University Hospital Duesseldorf  Germany"},{"name":"Liquid Biopsy Center Duesseldorf Center for Integrated Oncology Aachen  Bonn, Cologne Germany"},{"name":"Department of General, Visceral, Vascular and Transplantation Surgery University Hospital Essen  Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,9,18]]},"reference":[{"key":"e_1_2_11_2_1","unstructured":"Global cancer epidemiology situation.2026."},{"key":"e_1_2_11_3_1","unstructured":"National Cancer Institute \u2013 Surveillance E. and Results Program.Cancer stat facts: colorectal 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