Abstract 6035: Genetically-engineered mouse model for colonic neoplasia presenting mucinous and microsatellite unstable phenotype
Authors: Sada Haruki , Hinoi Takao , Niitsu Hiroaki , Miguchi Masashi , Sakamoto Naoya , Oue Naohide , Tashiro Hirotaka , Ohdan Hideki 
Published: June 2022

Background and Aim: Mucinous colorectal adenocarcinoma (CRC) is characterized by abundant extracellular mucin which occupies more than 50% of the tumor volume. Mucinous CRCs account for 10% of all CRCs and are likely to be microsatellite unstable including Lynch syndrome. Mucinous CRCs metastasize less frequently. However, once they metastasize, they exhibit resistance to cytotoxic chemotherapeutics, and are associated with poorer prognosis. Although there are some issues to be addressed in mucinous CRCs, mouse models are lacking for mucinous CRCs. Here, we report genetically-engineered mouse models (GEMMs) for mucin-producing colonic tumors with microsatellite instability (MSI), resembling human mucinous CRCs.

Method: To induce genetic alterations specifically in colonic epithelial cells, we used CDX2P-G19Cre transgenic mice. In this mouse model, a 9.5-kb fragment in the CDX2 promoter lesion (CDX2P9.5) regulates Cre expression specifically in colonic epithelium (Hinoi et al. Cancer Research 2007). In addition, 19 guanine nucleotides are inserted just after the first ATG, and Cre only expresses in the epithelial cells in which the number of the guanine tract stochastically turns a multiple of three. This mechanism helps our GEMMs to avoid embryonic death due to the genetic events we introduce. We have also reported that tumors in CDX2P9.5-G19Cre;Apcflox/flox mice have an abnormality in two of four MSI markers, suggesting that the tumors are driven by MSI (Sasada et al. PLoS One 2015). TGFBR2 mutation is frequently seen in human MSI CRCs, and inactivation of both Tgfrbr2 and Apc by Villin-Cre results in small intestinal neoplasm producing mucin. Therefore, to test if Tgfbr2 conditional knockout affects the mucin-producing phenotype in the context of MSI-high in this mouse model, we generated CDX2P-G19Cre;Apcflox/+;Tgfbr2flox/flox.

Results: In CDX2P-G19Cre; Apcflox/+; Tgfbr2 flox/flox mice, colonic neoplasia developed in 27 out of 40 mice. Approximately 60% of the tumors are well-differentiated adenocarcinoma producing mucin, while the remaining 40% are regular colonic adenomas. In these mucinous adenocarcinomas, tumor cells invade into the muscular layer. We also confirmed the immunoreactivity of β-catenin in cytoplasm and nuclei at the invasive front of the tumors in IHC analysis, indicating that these tumors are driven by abnormal Wnt activity. Nearby to the invasive front, there were mucin that were positive for a PAS reaction.

Conclusion: Mucin-producing adenocarcinomas develop in CDX2P9.5-G19Cre;Apcflox/+; Tgfbr2flox/flox mice. This GEMM is a potential mouse model recapitulating human mucinous and microsatellite unstable colorectal adenocarcinoma. The precise mechanism in which mucinous adenocarcinoma develops needs to be elucidated by genomic and transcriptomic analyses.

Citation Format: Haruki Sada, Takao Hinoi, Hiroaki Niitsu, Masashi Miguchi, Naoya Sakamoto, Naohide Oue, Hirotaka Tashiro, Hideki Ohdan. Genetically-engineered mouse model for colonic neoplasia presenting mucinous and microsatellite unstable phenotype [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6035.