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PET Imaging of l-Type Amino Acid Transporter (LAT1) and Cystine-Glutamate Antiporter (xc−) with [18F]FDOPA and [18F]FSPG in Breast Cancer Models

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Abstract

Purpose

The present study describes the analysis of amino acid transporters ASCT1, ASCT2, LAT1, and xc− in breast cancer under normoxic and hypoxic conditions. [18F]FDOPA-PET and [18F]FSPG-PET were used as imaging biomarkers to probe l-type amino acid transporter (LAT1) and cystine-glutamate antiporter (xc−) in breast cancer models.

Procedures

LAT1 and xc− transporters were studied under normoxic and hypoxic conditions with radiotracers [18F]FDOPA and [18F]FSPG in estrogen receptor–positive (ER+) MCF7 and triple-negative MDA-MB231 cells and in human mammary epithelial MCF10A control cells. Protein expression was analyzed using Western blot and immunohistochemistry.

Results

ASCT1 protein expression levels were comparable in all three cell lines, while noticeable ASCT2 expression levels were only found in MCF10A control cells. Higher LAT1 protein expression was detected in ER+ MCF7 cells. High xc− protein expression levels were detected in MDA-MB231 cells. Uptake of [18F]FDOPA through LAT1 was significantly higher in MCF7 versus MDA-MB231 cells, while the uptake of [18F]FSPG through xc− resulted in the opposite confirming expression and functional differences for both amino acid transporters in different breast cancer models. Hypoxia significantly increased [18F]FDOPA uptake in MCF7 cells and [18F]FSPG uptake in MDA-MB231 cells. In vivo PET imaging revealed substantially higher tumor uptake of [18F]FDOPA in MCF7 tumors as well as [18F]FSPG uptake in MDA-MB231 tumors confirming differences detected in vitro.

Conclusions

ER+ breast cancer cells express higher levels of amino acid transporter LAT1, whereas triple-negative breast cancer cells express more xc−. Cellular uptake and PET imaging experiments with [18F]FDOPA and [18F]FSPG confirmed functional LAT1 and xc− expression profiles. There was initial evidence that hypoxia regulates the function of both amino acid transporters in breast cancer. The results further indicated that [18F]FDOPA and [18F]FSPG are suitable radiotracer to distinguish between ER+ and triple-negative breast cancer types.

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Acknowledgments

The authors thank Dr. John Wilson, David Clendening, and Blake Lazurko from the Edmonton Radiopharmaceutical Center for 18F production as well as [18F]FDOPA production. The authors are also grateful to Dan McGinn (Vivarium of the Cross Cancer Institute, Edmonton, AB, Canada) for supporting the animal work and Dr. Hans-Soenke Jans (University of Alberta) for technical help and support of the PET imaging experiments. D.K. thanks the Alberta Cancer Foundation for a Graduate Student Scholarship and the Cancer Research Institute of Northern Alberta (CRINA) for la Vie en Rose Scholarship for Breast Cancer Research. The authors also gratefully acknowledge the Dianne and Irving Kipnes Foundation for supporting this work.

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Authors and Affiliations

  1. Department of Oncology, University of Alberta, 11560 University Ave, Edmonton, AB, T6G 1Z2, Canada

    Daniel Krys, Stephanie Mattingly, Darryl Glubrecht, Melinda Wuest & Frank Wuest

  2. Cancer Research Institute of Northern Alberta (CRINA), University of Alberta, Edmonton, AB, T6G 2S2, Canada

    Daniel Krys, Stephanie Mattingly, Melinda Wuest & Frank Wuest

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  1. Daniel Krys
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Correspondence to Frank Wuest.

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All animal experiments were carried out in accordance with guidelines of the Canadian Council on Animal Care (CCAC) and approved by the local animal care committee of the Cross Cancer Institute.

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Krys, D., Mattingly, S., Glubrecht, D. et al. PET Imaging of l-Type Amino Acid Transporter (LAT1) and Cystine-Glutamate Antiporter (xc−) with [18F]FDOPA and [18F]FSPG in Breast Cancer Models. Mol Imaging Biol 22, 1562–1571 (2020). https://doi.org/10.1007/s11307-020-01529-1

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