Mouse Models of Human Gastric Cancer Subtypes With Stomach-Specific CreERT2-Mediated Pathway Alterations

Therese Seidlitz; Yi-Ting Chen; Heike Uhlemann; Sebastian Schölch; Susan Kochall; Sebastian R Merker; Anna Klimova; Alexander Hennig; Christine Schweitzer; Kristin Pape; Gustavo B Baretton; Thilo Welsch; Daniela E Aust; Jürgen Weitz; Bon-Kyoung Koo; Daniel E Stange

doi:10.1053/j.gastro.2019.09.026

Mouse Models of Human Gastric Cancer Subtypes With Stomach-Specific CreERT2-Mediated Pathway Alterations

Gastroenterology. 2019 Dec;157(6):1599-1614.e2. doi: 10.1053/j.gastro.2019.09.026. Epub 2019 Oct 1.

Authors

Therese Seidlitz¹, Yi-Ting Chen², Heike Uhlemann¹, Sebastian Schölch³, Susan Kochall¹, Sebastian R Merker¹, Anna Klimova⁴, Alexander Hennig⁵, Christine Schweitzer¹, Kristin Pape¹, Gustavo B Baretton⁶, Thilo Welsch¹, Daniela E Aust⁶, Jürgen Weitz⁷, Bon-Kyoung Koo⁸, Daniel E Stange⁹

Affiliations

¹ Department of Visceral, Thoracic and Vascular Surgery, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany.
² Department of Pathology, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan; Department of Pathology, Faculty of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.
³ Department of Visceral, Thoracic and Vascular Surgery, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany; Department of Surgery, Universitätsmedizin Mannheim, Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany; German Cancer Consortium (DKTK), Dresden, Germany, and German Cancer Research Center (DKFZ), Heidelberg, Germany.
⁴ Institute for Medical Informatics and Biometry, Medical Faculty Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany; Core Unit for Data Management and Analytics (CDMA), National Center for Tumor Diseases (NCT), Dresden, Germany.
⁵ Department of Visceral, Thoracic and Vascular Surgery, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany; National Center for Tumor Diseases, Dresden, Germany.
⁶ German Cancer Consortium (DKTK), Dresden, Germany, and German Cancer Research Center (DKFZ), Heidelberg, Germany; Core Unit for Molecular Tumour Diagnostics, National Center for Tumor Diseases (NCT), Dresden, Germany; Institute of Pathology and Tumour and Normal Tissue Bank of the University Cancer Center, University Hospital Carl Gustav Carus, Medical Faculty, Technische Universität Dresden, Dresden, Germany; National Center for Tumor Diseases, Dresden, Germany.
⁷ Department of Visceral, Thoracic and Vascular Surgery, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany; German Cancer Consortium (DKTK), Dresden, Germany, and German Cancer Research Center (DKFZ), Heidelberg, Germany; National Center for Tumor Diseases, Dresden, Germany.
⁸ Institute of Molecular Biotechnology of the Austrian Academy of Sciences, Vienna Biocenter, Vienna, Austria.
⁹ Department of Visceral, Thoracic and Vascular Surgery, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany; German Cancer Consortium (DKTK), Dresden, Germany, and German Cancer Research Center (DKFZ), Heidelberg, Germany; National Center for Tumor Diseases, Dresden, Germany. Electronic address: daniel.stange@ukdd.de.

Abstract

Background & aims: Patterns of genetic alterations characterize different molecular subtypes of human gastric cancer. We aimed to establish mouse models of these subtypes.

Methods: We searched databases to identify genes with unique expression in the stomach epithelium, resulting in the identification of Anxa10. We generated mice with tamoxifen-inducible Cre recombinase (CreERT2) in the Anxa10 gene locus. We created 3 mouse models with alterations in pathways that characterize the chromosomal instability (CIN) and the genomically stable (GS) subtypes of human gastric cancer: Anxa10-CreER^T2;Kras^G12D/+;Tp53^R172H/+;Smad4^fl/f (CIN mice), Anxa10-CreER^T2;Cdh1^fl/fl;Kras^G12D/+;Smad4^fl/fl (GS-TGBF mice), and Anxa10-CreER^T2;Cdh1^fl/fl;Kras^G12D/+;Apc^fl/fl (GS-Wnt mice). We analyzed tumors that developed in these mice by histology for cell types and metastatic potential. We derived organoids from the tumors and tested their response to chemotherapeutic agents and the epithelial growth factor receptor signaling pathway inhibitor trametinib.

Results: The gastric tumors from the CIN mice had an invasive phenotype and formed liver and lung metastases. The tumor cells had a glandular morphology, similar to human intestinal-type gastric cancer. The gastric tumors from the GS-TGFB mice were poorly differentiated with diffuse morphology and signet ring cells, resembling human diffuse-type gastric cancer. Cells from these tumors were invasive, and mice developed peritoneal carcinomatosis and lung metastases. GS-Wnt mice developed adenomatous tooth-like gastric cancer. Organoids derived from tumors of GS-TGBF and GS-Wnt mice were more resistant to docetaxel, whereas organoids from the CIN tumors were more resistant to trametinib.

Conclusions: Using a stomach-specific CreERT2 system, we created mice that develop tumors with morphologic similarities to subtypes of human gastric cancer. These tumors have different patterns of local growth, metastasis, and response to therapeutic agents. They can be used to study different subtypes of human gastric cancer.

Keywords: Carcinogenesis; Cre/loxP System; Stomach Cancer; Targeted Therapy.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adenomatous Polyposis Coli Protein / genetics
Animals
Annexins / genetics
Antineoplastic Agents / pharmacology
Antineoplastic Agents / therapeutic use
Cell Transformation, Neoplastic / genetics
Disease Models, Animal*
Drug Resistance, Neoplasm / genetics
Female
Gastric Mucosa / pathology*
Genetic Loci / genetics*
Humans
Integrases / genetics
Male
Mice
Mice, Transgenic
Mutation
Proto-Oncogene Proteins p21(ras) / genetics
Smad4 Protein / genetics
Stomach Neoplasms / drug therapy
Stomach Neoplasms / genetics*
Stomach Neoplasms / pathology
Tumor Suppressor Protein p53 / genetics

Substances

Adenomatous Polyposis Coli Protein
Annexins
Antineoplastic Agents
Anxa10 protein, mouse
Smad4 Protein
Smad4 protein, mouse
Trp53 protein, mouse
Tumor Suppressor Protein p53
adenomatous polyposis coli protein, mouse
Cre recombinase
Integrases
Hras protein, mouse
Proto-Oncogene Proteins p21(ras)