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Inhibition of 6-phosphofructo-2-kinase (PFKFB3) suppresses glucose metabolism and the growth of HER2+ breast cancer

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Abstract

Purpose

Human epidermal growth factor receptor-2 (HER2) has been implicated in the progression of multiple tumor types, including breast cancer, and many downstream effectors of HER2 signaling are primary regulators of cellular metabolism, including Ras and Akt. A key downstream metabolic target of Ras and Akt is the 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 isozyme (PFKFB3), whose product, fructose-2,6-bisphosphate (F26BP), is a potent allosteric activator of a rate-limiting enzyme in glycolysis, 6-phosphofructo-1-kinase (PFK-1). We postulate that PFKFB3 may be regulated by HER2 and contribute to HER2-driven tumorigenicity.

Methods

Immunohistochemistry and Kaplan–Meier analysis of HER2+ patient samples investigated the relevance of PFKFB3 in HER2+ breast cancer. In vitro genetic and pharmacological inhibition of PFKFB3 was utilized to determine effects on HER2+ breast cancer cells, while HER2 antagonist treatment assessed the mechanistic regulation on PFKFB3 expression and glucose metabolism. Administration of a PFKFB3 inhibitor in a HER2-driven transgenic breast cancer model evaluated this potential therapeutic approach in vivo.

Results

PFKFB3 is elevated in human HER2+ breast cancer and high PFKFB3 transcript correlated with poorer progression-free (PFS) and distant metastatic-free (DFMS) survival. Constitutive HER2 expression led to elevated PFKFB3 expression and increased glucose metabolism, while inhibition of PFKFB3 suppressed glucose uptake, F26BP, glycolysis, and selectively decreased the growth of HER2-expressing breast cancer cells. In addition, treatment with lapatinib, an FDA-approved HER2 inhibitor, decreased PFKFB3 expression and glucose metabolism in HER2+ cells. In vivo administration of a PFKFB3 antagonist significantly suppressed the growth of HER2-driven breast tumors and decreased 18F-2-deoxy-glucose uptake.

Conclusions

Taken together, these data support the potential clinical utility of PFKFB3 inhibitors as chemotherapeutic agents against HER2+ breast cancer.

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Acknowledgments

This work was supported by the Department of Defense CDMRP (BC112204 to JO), the National Cancer Institute [CA166327 to BFC; CA149438 to JC], and by the American Cancer Society (RSG 13-139-01-CNE to BFC). We gratefully acknowledge Drs. Chin Ng and Huaiyu Zheng within the James Graham Brown Cancer Center (JGBCC) animal imaging core facility for assistance with the microCT and microPET procedures and Dr. Senthil Muthuswamy for providing the MCF10A-HER2 cells.

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Author notes

  1. Julie O’Neal

    Present address: Department of Medicine, Washington University, Saint Louis, MO, 63110, USA

Authors and Affiliations

  1. Department of Medicine, University of Louisville, Louisville, KY, 40202, USA

    Julie O’Neal, Amy Clem, Yoannis Imbert-Fernandez, Sucheta Telang & Jason Chesney

  2. Department of Biochemistry and Molecular Genetics, University of Louisville, 505 South Hancock Street, CTRB, Rm. 422, Louisville, KY, 40202, USA

    Lindsey Reynolds, Susan Dougherty & Brian F. Clem

  3. Molecular Targets Group, James Graham Brown Cancer Center, University of Louisville, Louisville, KY, 40202, USA

    Julie O’Neal, Amy Clem, Lindsey Reynolds, Susan Dougherty, Yoannis Imbert-Fernandez, Sucheta Telang, Jason Chesney & Brian F. Clem

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Correspondence to Brian F. Clem.

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Conflict of interest

ST, JC, and BFC are co-inventors on patents related to PFKFB3 inhibitors (US Patent#8,088,385) which are wholly owned by the University of Louisville Research Foundation.

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O’Neal, J., Clem, A., Reynolds, L. et al. Inhibition of 6-phosphofructo-2-kinase (PFKFB3) suppresses glucose metabolism and the growth of HER2+ breast cancer. Breast Cancer Res Treat 160, 29–40 (2016). https://doi.org/10.1007/s10549-016-3968-8

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