Pre-clinical imaging for establishment and comparison of orthotopic non-small cell lung carcinoma: in search for models reflecting clinical scenarios.

OBJECTIVE: Clinically relevant animal models of non-small cell lung carcinoma (NSCLC) are required for the validation of novel treatments. We compared two different orthotopic transplantation techniques as well as imaging modalities to identify suitable mouse models mimicking clinical scenarios. METHODS: We used three genomically diverse NSCLC cell lines [National Cancer Institute (NCI)-H1703 adenosquamous cell carcinoma, NCI-H23 adenocarcinoma and A549 adenocarcinoma) for implanting tumour cells either as spheroids or cell suspension into lung parenchyma. Bioluminescence imaging (BLI) and contrast-enhanced cone beam CT (CBCT) were performed twice weekly to monitor tumour growth. Tumour histological data and microenvironmental parameters were determined. RESULTS: Tumour development after spheroid-based transplantation differs probably due to the integrity of spheroids, as H1703 developed single localised nodules, whereas H23 showed diffuse metastatic spread starting early after transplantation. A549 transplantation as cell suspension with the help of a stereotactic system was associated with initial single localised tumour growth and eventual metastatic spread. Imaging techniques were successfully applied to monitor longitudinal tumour growth: BLI revealed highly sensitive qualitative data, whereas CBCT was associated with less sensitive quantitative data. Histology revealed significant model-dependent heterogeneity in proliferation, hypoxia, perfusion and necrosis. CONCLUSION: Our developed orthotopic NSCLC tumours have similarity with biological growth behaviour comparable to that seen in the clinic and could therefore be used as attractive models to study tumour biology and evaluate new therapeutic strategies. The use of human cancer cell lines facilitates testing of different genomic tumour profiles that may affect treatment outcomes. ADVANCES IN KNOWLEDGE: The combination of different imaging modalities to identify tumour growth with subsequent use in treatment planning and orthotopic transplantation techniques to develop initially single lesions to ultimate metastases pave the way towards representative pre-clinical NSCLC models for experimental testing of novel therapeutic options in future studies.

[1]  Robert M. Hoffman,et al.  Orthotopic Metastatic Mouse Models for Anticancer Drug Discovery and Evaluation: a Bridge to the Clinic , 2004, Investigational New Drugs.

[2]  R. Mak,et al.  Image-guided radiotherapy platform using single nodule conditional lung cancer mouse models , 2014, Nature Communications.

[3]  G. Giaccone,et al.  Reconstituted basement membrane (matrigel) and laminin can enhance the tumorigenicity and the drug resistance of small cell lung cancer cell lines. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[4]  G. Kayser,et al.  An orthotopic mouse model of small cell lung cancer reflects the clinical course in patients , 2016, Clinical & Experimental Metastasis.

[5]  Antje Dietrich,et al.  Precise image-guided irradiation of small animals: a flexible non-profit platform , 2016, Physics in medicine and biology.

[6]  T. Jacks,et al.  Imaging primary lung cancers in mice to study radiation biology. , 2010, International journal of radiation oncology, biology, physics.

[7]  R. Hoffman,et al.  Site-specific chemosensitivity of human small-cell lung carcinoma growing orthotopically compared to subcutaneously in SCID mice: the importance of orthotopic models to obtain relevant drug evaluation data. , 1993, Anticancer research.

[8]  W. Travis,et al.  New pathologic classification of lung cancer: relevance for clinical practice and clinical trials. , 2013, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[9]  T. Buckle,et al.  Validation of intratracheal instillation of lung tumour cells in mice using single photon emission computed tomography/computed tomography imaging , 2010, Laboratory animals.

[10]  Ido D. Weiss,et al.  In the Hunt for Therapeutic Targets: Mimicking the Growth, Metastasis, and Stromal Associations of Early-Stage Lung Cancer Using a Novel Orthotopic Animal Model , 2015, Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer.

[11]  Wolfgang Enghardt,et al.  Pre-clinical research in small animals using radiotherapy technology--a bidirectional translational approach. , 2014, Zeitschrift fur medizinische Physik.

[12]  M. Burt,et al.  Establishment of an experimental intrapulmonary tumor nodule model. , 1997, The Annals of thoracic surgery.

[13]  J. Carlsson,et al.  Liquid-overlay culture of cellular spheroids. , 1984, Recent results in cancer research. Fortschritte der Krebsforschung. Progres dans les recherches sur le cancer.

[14]  I. Fidler,et al.  Orthotopic Models are Necessary to Predict Therapy of Transplantable Tumors in Mice , 1998, Cancer and Metastasis Reviews.

[15]  A. Sweet-Cordero,et al.  Hypoxia in Models of Lung Cancer: Implications for Targeted Therapeutics , 2010, Clinical Cancer Research.

[16]  B. Wilson,et al.  Orthotopic lung cancer murine model by nonoperative transbronchial approach. , 2014, The Annals of thoracic surgery.

[17]  H. Kalthoff,et al.  An improved orthotopic xenotransplant procedure for human lung cancer in SCID bg mice. , 2000, The Annals of thoracic surgery.

[18]  liyan Huang,et al.  Establishment of an orthotopic lung cancer model in nude mice and its evaluation by spiral CT. , 2012, Journal of thoracic disease.

[19]  M. Bibby,et al.  Orthotopic models of cancer for preclinical drug evaluation: advantages and disadvantages. , 2004, European journal of cancer.

[20]  Y. Castier,et al.  Bioluminescent Orthotopic Mouse Models of Human Localized Non-Small Cell Lung Cancer: Feasibility and Identification of Circulating Tumour Cells , 2011, PloS one.

[21]  H. Dosaka-akita,et al.  Prognostic significance of p27KIP1 protein and ki-67 growth fraction in non-small cell lung cancers. , 2000, Clinical cancer research : an official journal of the American Association for Cancer Research.

[22]  Hassan Lemjabbar-Alaoui,et al.  Lung cancer: Biology and treatment options. , 2015, Biochimica et biophysica acta.

[23]  P. Bunn,et al.  Irradiated nude rat model for orthotopic human lung cancers. , 1991, Cancer research.

[24]  A. Berns,et al.  Induction of small cell lung cancer by somatic inactivation of both Trp53 and Rb1 in a conditional mouse model. , 2003, Cancer cell.

[25]  R. Herbst,et al.  Development of an orthotopic model to study the biology and therapy of primary human lung cancer in nude mice. , 2003, Clinical cancer research : an official journal of the American Association for Cancer Research.

[26]  Timothy D Solberg,et al.  An Orthotopic Lung Tumor Model for Image-Guided Microirradiation in Rats , 2010, Radiation research.

[27]  A. Jemal,et al.  Cancer statistics, 2017 , 2017, CA: a cancer journal for clinicians.

[28]  M. Krause,et al.  Pimonidazole labelling and response to fractionated irradiation of five human squamous cell carcinoma (hSCC) lines in nude mice: the need for a multivariate approach in biomarker studies. , 2006, Radiotherapy and oncology : journal of the European Society for Therapeutic Radiology and Oncology.

[29]  L. Kraus-Berthier,et al.  Histology and sensitivity to anticancer drugs of two human non-small cell lung carcinomas implanted in the pleural cavity of nude mice. , 2000, Clinical cancer research : an official journal of the American Association for Cancer Research.

[30]  C. Contag,et al.  Advancing animal models of neoplasia through in vivo bioluminescence imaging. , 2002, European journal of cancer.

[31]  N. Heuzé-Vourc’h,et al.  Monitoring of tumour progression using bioluminescence imaging and computed tomography scanning in a nude mouse orthotopic model of human small cell lung cancer. , 2012, Lung cancer.

[32]  R. Bjerkvig,et al.  In Vivo Bioluminescence Imaging Validation of a Human Biopsy–Derived Orthotopic Mouse Model of Glioblastoma Multiforme , 2013, Molecular imaging.

[33]  Juergen Friedrich,et al.  Spheroid-based drug screen: considerations and practical approach , 2009, Nature Protocols.

[34]  Jos Jonkers,et al.  Genetically engineered mouse models in oncology research and cancer medicine , 2016, EMBO molecular medicine.