Characterization Of A Novel Transplantable C3tag Breast Cancer Model In C57bl/6j Mice.

Abstract Introduction: In women in the United States, breast cancer (BC) is the second leading cause of cancer-related death. BC is a heterogeneous disease, characterized by several subtypes based on molecular markers including several hormone receptors. Transcriptomic analysis further stratifies the disease. The basal-like subtype, often triple-negative for hormone receptors, has the poorest prognosis partially due to the absence of targeted therapies. Obesity, a preventable cause of cancer, is an established risk factor for breast cancer, including basal-like breast cancers. Mouse models to study the interaction of BC and obesity, particularly the more aggressive basal-like BC, are crucial. The C3(1)/SV40 large T-antigen (TAg) transgenic mouse model is an established model of basal-like mammary cancer that recapitulates several key features of human breast cancer. However, this model was generated in FVB mice, which is not an ideal genetic background for studying obesity, immunology and other aspects of cancer research. The goal of this study was to develop and characterize a C3TAg transgenic mouse model on a C57BL/6 background. Therefore, the current study backcrossed C3TAg transgenic FVB mice with C57BL/6 mice, a model most commonly used to study obesity, for 10 generations to create a transplantable C3TAg model in C57BL/6 mice. Methods: Cell Line Generation: C3TAg transgenic FVB mice were backcrossed with C57BL/6 mice for 10 generations. Tumors from 6-month-old heterozygous female mammary tissue were harvested, dissociated, and subcloned by limited dilution. A total of 156 clonal lines were established from which a subset of 12 lines displaying differences in morphology, growth and expression of metabolic and EMT genes (by qPCR). In vivo: 96 female C57BL/6J mice were randomized into 12 groups (n=8/group) and orthotopically injected with three different cell densities (5*104, 1.5*105, 5*105 cells/mouse) from one of the four selected lines. Mice tumor growth was assessed using electronic calipers. In vitro: Proliferation and cellular respiration were assessed and molecular subtyping was conducted using Affymetrix microarray data. Results: All tested cell lines formed tumors in vivo, with macroscopic characteristics of basal-like tumors including central necrosis. In vitro growth and metabolism of tested lines showed modest dissimilarity; in contrast, tumor growth rates and metastatic outgrowth were starkly dissimilar among the tested lines. Transcriptomic data clustered these cell lines with human BC tumors. Conclusions: Here we report for the first time a transplantable C3TAg model in C57BL/6 mice with highly effective tumor engraftment and growth, and with variable phenotypes for tumor growth and metastasis. Thus, this model may serve as an attractive model for the study of the interactions of basal-like breast cancer and obesity. This work was supported by R35CA197627 to SH. Citation Format: Meredith S. Carson, Michael F. Colman, Daniel Roth, Jody Albright, Melissa VerHague, John E. French, Stephen D. Hursting. Characterization of a novel transplantable C3TAg breast cancer model in C57BL/6J mice [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2918.