Organoids and metastatic orthotopic mouse model for mismatch repair-deficient colorectal cancer

Yurong Song; Travis D Kerr; Chelsea Sanders; Lisheng Dai; Shaneen S Baxter; Brandon Somerville; Ryan N Baugher; Stephanie D Mellott; Todd B Young; Heidi E Lawhorn; Teri M Plona; Bingfang Xu; Lei Wei; Qiang Hu; Song Liu; Alan Hutson; Baktiar Karim; Sandra Burkett; Simone Difilippantonio; Ligia Pinto; Johannes Gebert; Matthias Kloor; Steven M Lipkin; Shizuko Sei; Robert H Shoemaker

doi:10.3389/fonc.2023.1223915

Organoids and metastatic orthotopic mouse model for mismatch repair-deficient colorectal cancer

Front Oncol. 2023 Sep 8:13:1223915. doi: 10.3389/fonc.2023.1223915. eCollection 2023.

Authors

Yurong Song¹, Travis D Kerr¹, Chelsea Sanders², Lisheng Dai¹, Shaneen S Baxter¹, Brandon Somerville¹, Ryan N Baugher³, Stephanie D Mellott³, Todd B Young³, Heidi E Lawhorn³, Teri M Plona³, Bingfang Xu⁴, Lei Wei⁵, Qiang Hu⁵, Song Liu⁵, Alan Hutson⁵, Baktiar Karim⁶, Sandra Burkett⁷, Simone Difilippantonio², Ligia Pinto¹, Johannes Gebert⁸, Matthias Kloor⁸, Steven M Lipkin⁹, Shizuko Sei¹⁰, Robert H Shoemaker¹⁰

Affiliations

¹ Frederick National Laboratory for Cancer Research, Vaccine, Immunity, and Cancer Directorate, Frederick, MD, United States.
² Frederick National Laboratory for Cancer Research, Laboratory Animal Sciences Program, Frederick, MD, United States.
³ Frederick National Laboratory for Cancer Research, Clinical Laboratory Improvement Amendments (CLIA) Molecular Diagnostics Laboratory, Frederick, MD, United States.
⁴ Frederick National Laboratory for Cancer Research, Genomics Laboratory, Frederick, MD, United States.
⁵ Department of Biostatistics and Bioinformatics, Roswell Park Comprehensive Cancer Center, Buffalo, NY, United States.
⁶ Molecular Histopathology Laboratory, Frederick National Laboratory for Cancer Research, Frederick, MD, United States.
⁷ Molecular Cytogenetics Core Facility, National Cancer Institute, Frederick, MD, United States.
⁸ Department of Applied Tumor Biology, Institute of Pathology, University of Heidelberg, Heidelberg, Germany.
⁹ Department of Medicine, Weill Cornell Medical College, Cornell University, New York, NY, United States.
¹⁰ Chemopreventive Agent Development Research Group, Division of Cancer Prevention, National Cancer Institute, Bethesda, MD, United States.

Abstract

Background: Genome integrity is essential for the survival of an organism. DNA mismatch repair (MMR) genes (e.g., MLH1, MSH2, MSH6, and PMS2) play a critical role in the DNA damage response pathway for genome integrity maintenance. Germline mutations of MMR genes can lead to Lynch syndrome or constitutional mismatch repair deficiency syndrome, resulting in an increased lifetime risk of developing cancer characterized by high microsatellite instability (MSI-H) and high mutation burden. Although immunotherapy has been approved for MMR-deficient (MMRd) cancer patients, the overall response rate needs to be improved and other management options are needed.

Methods: To better understand the biology of MMRd cancers, elucidate the resistance mechanisms to immune modulation, and develop vaccines and therapeutic testing platforms for this high-risk population, we generated organoids and an orthotopic mouse model from intestine tumors developed in a Msh2-deficient mouse model, and followed with a detailed characterization.

Results: The organoids were shown to be of epithelial origin with stem cell features, to have a high frameshift mutation frequency with MSI-H and chromosome instability, and intra- and inter-tumor heterogeneity. An orthotopic model using intra-cecal implantation of tumor fragments derived from organoids showed progressive tumor growth, resulting in the development of adenocarcinomas mixed with mucinous features and distant metastasis in liver and lymph node.

Conclusions: The established organoids with characteristics of MSI-H cancers can be used to study MMRd cancer biology. The orthotopic model, with its distant metastasis and expressing frameshift peptides, is suitable for evaluating the efficacy of neoantigen-based vaccines or anticancer drugs in combination with other therapies.

Keywords: Lynch syndrome; MSH2; chromosome instability; colorectal cancer; microsatellite instability; mismatch repair deficiency; mouse model; organoid.

Copyright © 2023 Song, Kerr, Sanders, Dai, Baxter, Somerville, Baugher, Mellott, Young, Lawhorn, Plona, Xu, Wei, Hu, Liu, Hutson, Karim, Burkett, Difilippantonio, Pinto, Gebert, Kloor, Lipkin, Sei and Shoemaker.

Grants and funding

This research was supported in part with federal funds from the National Cancer Institute, National Institutes of Health, under Contract No. HHSN261201500003I; IOTN Moonshot grant U24CA232979-01 (LW, QH, SL, and AH); and NIH grants U54 CA 272688U01 and U01 CA233056 and contracts from NCI PREVENT program (HHSN2612015000391) (SML, JG, and MK). The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the U.S. government.