title: Development of bioluminescent chick chorioallantoic membrane (CAM) models for primary pancreatic cancer cells: a platform for drug testing.
abstract: The aim of the present study was to develop chick-embryo chorioallantoic membrane (CAM) bioluminescent tumor models employing low passage cell cultures obtained from primary pancreatic ductal adenocarcinoma (PDAC) cells. Primary PDAC cells transduced with lentivirus expressing Firefly-luciferase (Fluc) were established and inoculated onto the CAM membrane, with >80% engraftment. Fluc signal reliably correlated with tumor growth. Tumor features were evaluated by immunohistochemistry and genetic analyses, including analysis of mutations and mRNA expression of PDAC pivotal genes, as well as microRNA (miRNA) profiling. These studies showed that CAM tumors had histopathological and genetic characteristic comparable to the original tumors. We subsequently tested the modulation of key miRNAs and the activity of gemcitabine and crizotinib on CAM tumors, showing that combination treatment resulted in 63% inhibition of tumor growth as compared to control (p < 0.01). These results were associated with reduced expression of miR-21 and increased expression of miR-155. Our study provides the first evidence that transduced primary PDAC cells can form tumors on the CAM, retaining several histopathological and (epi)genetic characteristics of original tumors. Moreover, our results support the use of these models for drug testing, providing insights on molecular mechanisms underlying antitumor activity of new drugs/combinations.
authors.affiliation: Department of Medical Oncology, Cancer Center Amsterdam, VU University Medical Center, Amsterdam, The Netherlands.|Department of Internal Medicine, Agios Nikolaos General Hospital, Agios Nikolaos, Crete, Greece.
authors.forename: Maria
authors.initials: M
authors.lastname: Rovithi
authors.affiliation: Molecular Medicine Group, Department of Modern Sciences and Technologies; School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: Amir
authors.initials: A
authors.lastname: Avan
authors.affiliation: Cancer Pharmacology Lab, AIRC Start-Up Unit, University of Pisa, Pisa, Italy.
authors.forename: Niccola
authors.initials: N
authors.lastname: Funel
authors.affiliation: Cancer Pharmacology Lab, AIRC Start-Up Unit, University of Pisa, Pisa, Italy.
authors.forename: Leticia G
authors.initials: LG
authors.lastname: Leon
authors.affiliation: Department of Medical Oncology, Cancer Center Amsterdam, VU University Medical Center, Amsterdam, The Netherlands.
authors.forename: Valentina E
authors.initials: VE
authors.lastname: Gomez
authors.affiliation: Department of Neurosurgery, Cancer Center Amsterdam, VU University Medical Center, Amsterdam, The Netherlands.|Molecular Neurogenetics Unit, Department of Neurology, Massachusetts General Hospital and Neuroscience Program, Harvard Medical School, Boston, Massachusetts, US.
authors.forename: Thomas
authors.initials: T
authors.lastname: Wurdinger
authors.affiliation: Department of Medical Oncology, Cancer Center Amsterdam, VU University Medical Center, Amsterdam, The Netherlands.
authors.forename: Arjan W
authors.initials: AW
authors.lastname: Griffioen
authors.affiliation: Department of Medical Oncology, Cancer Center Amsterdam, VU University Medical Center, Amsterdam, The Netherlands.
authors.forename: Henk M W
authors.initials: HM
authors.lastname: Verheul
authors.affiliation: Department of Medical Oncology, Cancer Center Amsterdam, VU University Medical Center, Amsterdam, The Netherlands.|Cancer Pharmacology Lab, AIRC Start-Up Unit, University of Pisa, Pisa, Italy.
authors.forename: Elisa
authors.initials: E
authors.lastname: Giovannetti
mesh_terms: D000230:Adenocarcinoma / Q000188:drug therapy / Q000235:genetics / Q000473:pathology; D000818:Animals; D063646:Carcinogenesis / Q000473:pathology; D021441:Carcinoma, Pancreatic Ductal / Q000188:drug therapy / Q000235:genetics / Q000473:pathology; D049109:Cell Proliferation / Q000187:drug effects / Q000235:genetics; D002645:Chickens; D049033:Chorioallantoic Membrane / Q000378:metabolism*; D000077547:Crizotinib / Q000494:pharmacology / Q000627:therapeutic use; D004252:DNA Mutational Analysis; D003841:Deoxycytidine / Q000031:analogs & derivatives / Q000494:pharmacology / Q000627:therapeutic use; D004353:Drug Evaluation, Preclinical; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D051559:Hepatocyte Nuclear Factor 6 / Q000235:genetics / Q000378:metabolism; D008163:Luminescent Measurements / Q000379:methods*; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism; D008954:Models, Biological; D010190:Pancreatic Neoplasms / Q000188:drug therapy / Q000235:genetics / Q000473:pathology*; D055758:SOX9 Transcription Factor / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D014407:Tumor Cells, Cultured; D000093542:Gemcitabine
chemical_list: D051559:Hepatocyte Nuclear Factor 6; D035683:MicroRNAs; D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human; D003841:Deoxycytidine; D000077547:Crizotinib; D000093542:Gemcitabine