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Cellular and Molecular Biology

Metabolic stress induces GD2+ cancer stem cell-like phenotype in triple-negative breast cancer

British Journal of Cancer volume 126, pages 615–627 (2022)Cite this article

Subjects

Abstract

Background

Metabolic stress resulting from nutrient deficiency is one of the hallmarks of a growing tumour. Here, we tested the hypothesis that metabolic stress induces breast cancer stem-like cell (BCSC) phenotype in triple-negative breast cancer (TNBC).

Methods

Flow cytometry for GD2 expression, mass spectrometry and Ingenuity Pathway Analysis for metabolomics, bioinformatics, in vitro tumorigenesis and in vivo models were used.

Results

Serum/glucose deprivation not only increased stress markers but also enhanced GD2+ BCSC phenotype and function in TNBC cells. Global metabolomics profiling identified upregulation of glutathione biosynthesis in GD2high cells, suggesting a role of glutamine in the BCSC phenotype. Cueing from the upregulation of the glutamine transporters in primary breast tumours, inhibition of glutamine uptake using small-molecule inhibitor V9302 reduced GD2+ cells by 70–80% and BCSC characteristics in TNBC cells. Mechanistic studies revealed inhibition of the mTOR pathway and induction of ferroptosis by V9302 in TNBC cells. Finally, inhibition of glutamine uptake significantly reduced in vivo tumour growth in a TNBC patient-derived xenograft model using NSG (non-obese diabetic/severe combined immunodeficiency with a complete null allele of the IL-2 receptor common gamma chain) mice.

Conclusion

Here, we show metabolic stress results in GD2+ BCSC phenotype in TNBC and glutamine contributes to GD2+ phenotype, and targeting the glutamine transporters could complement conventional chemotherapy in TNBC.

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Fig. 1: Serum and glucose deprivation induces GD2 expression in TNBC.
Fig. 2: Metabolomic profiling of breast cancer stem cells.
Fig. 3: METABRIC dataset show higher expression of SLC1A5 in breast cancer patients and is associated with poor survivability.
Fig. 4: V9302 inhibits GD2 expression, BCSC function and the glutathione (GSH) pathway in TNBC cells.
Fig. 5: V9302 induces synergistic killing and ferroptosis and inhibits the mTOR pathway in TNBC cells.
Fig. 6: Inhibition of glutamine metabolism reduces in vivo tumorigenesis.

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Acknowledgements

We are grateful to the patients who provided tumour biopsies for PDX model establishment. PDX models and derivatives were obtained from the Cazalot Breast Cancer Model Resource at The University of Texas MD Anderson Cancer Center. This resource was established through a gift from the Cazalot family and from funds from the MD Anderson Cancer Center Breast Cancer Moon Shot Programme. Editorial support was provided by Sunita Patterson and Bryan Tutt in Editing Services, Research Medical Library, MD Anderson Cancer Center. This study was supported by the Department of Defense (DOD), Grant #BC181493 (to VLB) and a Breast Cancer Research Foundation (BCRF) grant (to MA). Metabolomics and data analysis were performed by Metabolomics core facility, Baylor College of Medicine. This project was supported by CPRIT Proteomics and Metabolomics Core Facility (to NP), (RP170005), National Institute of Health (NIH)/National Cancer Institute (NCI) grant (P30 CA125123), Dan L. Duncan Cancer Center and NIH/NCI R01CA220297 (to NP) and NIH/NCI R01CA216426 (to NP). Additional funding sources that supported this work include Cancer Prevention and Research Institute of Texas grants RP150148 (to HP-W).

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Author notes

  1. These authors contributed equally: Appalaraju Jaggupilli, Stanley Ly.

Authors and Affiliations

  1. Section of Molecular Hematology and Therapy, Department of Leukemia, The University of Texas MD Anderson Cancer Center, Houston, TX, USA

    Appalaraju Jaggupilli, Stanley Ly, Khoa Nguyen, Vivek Anand, Bin Yuan, Fouad El-Dana, Zoe Arvanitis, Michael Andreeff & V. Lokesh Battula

  2. Department of Neurosurgery, McGovern Medical School, The University of Texas Health Science Center, Houston, TX, USA

    Yuanqing Yan

  3. Department of Molecular and Cell Biology, Baylor College of Medicine, Houston, TX, USA

    Danthasinghe Waduge Badrajee Piyarathna & Nagireddy Putluri

  4. Experimental Radiation Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA

    Helen Piwnica-Worms

  5. Center for Molecular Probes, Vanderbilt University Institute of Imaging Science, Nashville, TN, USA

    Henry Charles Manning

  6. Section of Translational Breast Cancer Research, Department of Breast Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA

    V. Lokesh Battula

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  1. Appalaraju Jaggupilli
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Contributions

AJ and VLB conceived the ideas, designed experiments, interpreted data and wrote the manuscript with inputs from all authors. AJ, SL and KN performed a majority of the experiments. BY assisted with Western blot and animal experiments. VA performed flow cytometry, qPCR and mammosphere assays. FE-D and ZA assisted with qPCR and mammosphere assays. DWBP and NP performed experiments and analyses pertaining to metabolomics. YY performed bioinformatics and combination index analysis. HP-W contributed to TNBC PDX models. HCM contributed V9302. MA and VLB supervised and directed all aspects of the study.

Corresponding author

Correspondence to V. Lokesh Battula.

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The authors declare no competing interests.

Ethics approval and consent to participate

This study was performed in accordance with the Declaration of Helsinki. All experiments involving animals were approved by and conducted in accordance with the policies of the Institutional Animal Care and Use Committee (IACUC) of The University of Texas MD Anderson Cancer Center.

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Jaggupilli, A., Ly, S., Nguyen, K. et al. Metabolic stress induces GD2+ cancer stem cell-like phenotype in triple-negative breast cancer. Br J Cancer 126, 615–627 (2022). https://doi.org/10.1038/s41416-021-01636-y

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