Abstract PR03: GLUT1 is required for induction of mammary tumorigenesis by activated ErbB2/HER2/Neu
Authors: Wellberg Elizabeth A. , D'Allessandro Angelo , Lewis Andrew S. , Terrell Kristina , Abel E. Dale , Muller William A. , Hansen Kirk A. , Anderson Steven M. 
Published: May 2016

Alterations in tumor cell metabolism have been investigated for decades, and include changes in glucose utilization, as well as changes in mitochondrial and fatty acid metabolism; however, it is unclear at what point during the process of tumorigenesis these metabolic changes occur. We hypothesized that elevated glucose uptake represents the first metabolic adaptation to transformation, and thus may be a target for the prevention of breast cancer. The GLUT1 transporter is expressed in breast cancer cells and is responsible for the majority of their glucose uptake. Not surprisingly, breast cancer patients with high levels of GLUT1 have a poorer clinical outcome than those with lower levels, likely due to the association between glucose uptake and high tumor grade. We previously showed that the reduction of GLUT1 in established tumor cells resulted in a 50% decrease in glucose consumption and lactate generation. There was a corresponding decrease in proliferation in both two and three-dimensional culture, and in tumor growth in immunodeficient mice. Mammary epithelial cells from GLUT1fl/fl mice were transformed using polyoma middle tumor antigen (PyMT), and GLUT1 excised using Cre recombinase. Cells expressing GLUT1 were tumorigenic in immunodeficient mice, while cells lacking GLUT1 were not tumorigenic. This suggests that in mammary epithelial cells transformed in vitro, loss of GLUT1 was sufficient to prevent tumor outgrowths when injected into the mammary fat pad of nude mice. This indicates that GLUT1 and by extension glucose, are critical at an early stage of mammary tumorigenesis.

To test the hypothesis that elevated glucose uptake is required very early in the process of tumor formation, we crossed the MMTV-NIC (NIC) mice, which contain activated NeuNT, an internal ribosome entry site (IRES), and Cre recombinase, to mice bearing floxed alleles of GLUT1, and monitored tumor development. All of the control MMTV-NIC mice developed tumors in less than 200 days. In contrast, none of the NIC-GLUT1fl/fl mice developed mammary tumors even after eighteen months. We anticipated that loss of one allele of GLUT1 would significantly delay mammary tumorigenesis; however, we did not observe any tumors in these mice, even at eighteen months of age. Tumors that formed in the NIC mice were typical Neu-dependent tumors observed in similar transgenic mice with a high proliferative index (38% by Ki67 staining), and there was evidence of preneoplastic lesions in the non tumor-bearing mammary glands. Small Cre-positive lesions were also positive for GLUT1 indicating GLUT1 expression increases very early in Neu-mediated tumorigenesis. The mammary ductal tree developed normally in NIC-Glut1fl/fl and NIC-Glut1fl/+ mice. Mammary glands from mice lacking one allele of GLUT1 had increased epithelial content and higher proliferative index, compared to mice lacking both Glut1 alleles; however, no palpable tumors ever formed in either group. These results suggest that loss of a single allele of GLUT1 is sufficient to suppress Neu-induced mammary tumorigenesis and that there is an absolute requirement for GLUT1 at the earliest stages of tumorigenesis.

Citation Format: Elizabeth A. Wellberg, Angelo D'Allessandro, Andrew S. Lewis, Kristina Terrell, E. Dale Abel, William A. Muller, Kirk A. Hansen, Steven M. Anderson. GLUT1 is required for induction of mammary tumorigenesis by activated ErbB2/HER2/Neu. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Breast Cancer Research; Oct 17-20, 2015; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(2_Suppl):Abstract nr PR03.