Cancer Metabolism

Enhanced glutamine uptake influences composition of immune cell infiltrates in breast cancer



Cancer cells must alter their metabolism to support proliferation. Immune evasion also plays a role in supporting tumour progression. This study aimed to find whether enhanced glutamine uptake in breast cancer (BC) can derive the existence of specific immune cell subtypes, including the subsequent impact on patient outcome.


SLC1A5, SLC7A5, SLC3A2 and immune cell markers CD3, CD8, FOXP3, CD20 and CD68, in addition to PD1 and PDL1, were assessed by using immunohistochemistry on TMAs constructed from a large BC cohort (n = 803). Patients were stratified based on SLC protein expression into accredited clusters and correlated with immune cell infiltrates and patient outcome. The effect of transient siRNA knockdown of SLC7A5 and SLC1A5 on PDL1 expression was evaluated in MDA-MB-231 cells.


High SLCs were significantly associated with PDL1 and PD1 +, FOXP3 +, CD68 + and CD20 + cells (p < 0.001). Triple negative (TN), HER2 + and luminal B tumours showed variable associations between SLCs and immune cell types (p ≤ 0.04). The expression of SLCs and PDL1, PD1 +, FOXP3 + and CD68 + cells was associated with poor patient outcome (p < 0.001). Knockdown of SLC7A5 significantly reduced PDL1 expression.


This study provides data that altered glutamine pathways in BC that appears to play a role in deriving specific subtypes of immune cell infiltrates, which either support or counteract its progression.

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  1. 1.

    Hanahan, D. & Weinberg, R. A. Hallmarks of cancer: the next generation. Cell 144, 646–674 (2011).

  2. 2.

    Bar-Peled, L. & Sabatini, D. M. Regulation of mTORC1 by amino acids. Trends Cell Biol. 24, 400–406 (2014).

  3. 3.

    Bond, P. Regulation of mTORC1 by growth factors, energy status, amino acids and mechanical stimuli at a glance. J. Int. Soc. Sports Nutr. 13, 8 (2016).

  4. 4.

    Bhutia, Y. D., Babu, E., Ramachandran, S. & Ganapathy, V. Amino acid transporters in cancer and their relevance to “glutamine addiction”: novel targets for the design of a new class of anticancer drugs. Cancer Res. 75, 1782–1788 (2015).

  5. 5.

    Fuchs, B. C. & Bode, B. P. Amino acid transporters ASCT2 and LAT1 in cancer: partners in crime? Semin. Cancer Biol. 15, 254–266 (2005).

  6. 6.

    Yanagida, O., Kanai, Y., Chairoungdua, A., Kim, D. K., Segawa, H., Nii, T. et al. Human L-type amino acid transporter 1 (LAT1): characterization of function and expression in tumor cell lines. Biochim, Biophys, Acta. 1514, 291–302 (2001).

  7. 7.

    El Ansari, R., Craze, M. L., Miligy, I., Diez-Rodriguez, M., Nolan, C. C., Ellis, I. O. et al. The amino acid transporter SLC7A5 confers a poor prognosis in the highly proliferative breast cancer subtypes and is a key therapeutic target in luminal B tumours. Breast Cancer Res. 20, 21 (2018).

  8. 8.

    El Ansari, R., Craze, M. L., Diez-Rodriguez, M., Nolan, C. C., Ellis, I. O., Rakha, E. A. et al. The multifunctional solute carrier 3A2 (SLC3A2) confers a poor prognosis in the highly proliferative breast cancer subtypes. Br. J. Cancer 118, 1115 (2018).

  9. 9.

    El-Ansari, R., Craze, M. L., Alfarsi, L., Soria, D., Diez-Rodriguez, M., Nolan, C. C. et al. The combined expression of solute carriers is associated with a poor prognosis in highly proliferative ER+ breast cancer. Breast Cancer Res. Tr. 175, 27–38 (2019).

  10. 10.

    Monjazeb, A. M., Zamora, A. E., Grossenbacher, S. K., Mirsoian, A., Sckisel, G. D. & Murphy, W. J. Immunoediting and antigen loss: overcoming the achilles heel of immunotherapy with antigen non-specific therapies. Front. Oncol. 3, 197 (2013).

  11. 11.

    Qian, B. Z. & Pollard, J. W. Macrophage diversity enhances tumor progression and metastasis. Cell 141, 39–51 (2010).

  12. 12.

    Vinay, D. S., Ryan, E. P., Pawelec, G., Talib, W. H., Stagg, J., Elkord, E. et al. Immune evasion in cancer: mechanistic basis and therapeutic strategies. Semin. Cancer Biol. 35(Suppl), S185–S98. (2015).

  13. 13.

    Mahmoud, S. M., Paish, E. C., Powe, D. G., Macmillan, R. D., Grainge, M. J., Lee, A. H. et al. Tumor-infiltrating CD8+ lymphocytes predict clinical outcome in breast cancer. J. Clin. Oncol. 29, 1949–1955 (2011).

  14. 14.

    Mahmoud, S. M., Paish, E. C., Powe, D. G., Macmillan, R. D., Lee, A. H., Ellis, I. O. et al. An evaluation of the clinical significance of FOXP3+ infiltrating cells in human breast cancer. Breast Cancer Res. Tr. 127, 99–108 (2011).

  15. 15.

    Mahmoud, S. M., Lee, A. H., Paish, E. C., Macmillan, R. D., Ellis, I. O. & Green, A. R. The prognostic significance of B lymphocytes in invasive carcinoma of the breast. Breast Cancer Res. Tr. 132, 545–553 (2012).

  16. 16.

    Konishi, J., Yamazaki, K., Azuma, M., Kinoshita, I., Dosaka-Akita, H. & Nishimura, M. B7-H1 expression on non-small cell lung cancer cells and its relationship with tumor-infiltrating lymphocytes and their PD-1 expression. Clinical Cancer Res. 10, 5094–5100 (2004).

  17. 17.

    Nomi, T., Sho, M., Akahori, T., Hamada, K., Kubo, A., Kanehiro, H. et al. Clinical significance and therapeutic potential of the programmed death-1 ligand/programmed death-1 pathway in human pancreatic cancer. Clinical Cancer Res. 13, 2151–2157 (2007).

  18. 18.

    Hamanishi, J., Mandai, M., Iwasaki, M., Okazaki, T., Tanaka, Y., Yamaguchi, K. et al. Programmed cell death 1 ligand 1 and tumor-infiltrating CD8+ T lymphocytes are prognostic factors of human ovarian cancer. Proc. Natl Acad. Sci. USA 104, 3360–3365 (2007).

  19. 19.

    Badoual, C., Hans, S., Merillon, N., Van Ryswick, C., Ravel, P., Benhamouda, N. et al. PD-1-expressing tumor-infiltrating T cells are a favorable prognostic biomarker in HPV-associated head and neck cancer. Cancer Res. 73, 128–138 (2013).

  20. 20.

    Sabatier, R., Finetti, P., Mamessier, E., Adelaide, J., Chaffanet, M., Ali, H. R. et al. Prognostic and predictive value of PDL1 expression in breast cancer. Oncotarget. 6, 5449–5464 (2015).

  21. 21.

    Kareva, I. & Hahnfeldt, P. The emerging “hallmarks” of metabolic reprogramming and immune evasion: distinct or linked? Cancer Res. 73, 2737–2742 (2013).

  22. 22.

    Carmona-Fontaine, C., Bucci, V., Akkari, L., Deforet, M., Joyce, J. A. & Xavier, J. B. Emergence of spatial structure in the tumor microenvironment due to the Warburg effect. Proc. Natl Acad. Sci. USA 110, 19402–19407 (2013).

  23. 23.

    Abd El-Rehim, D. M., Ball, G., Pinder, S. E., Rakha, E., Paish, C., Robertson, J. F. et al. High-throughput protein expression analysis using tissue microarray technology of a large well-characterised series identifies biologically distinct classes of breast cancer confirming recent cDNA expression analyses. Int. J. Cancer 116, 340–350 (2005).

  24. 24.

    Green, A. R., Aleskandarany, M. A., Ali, R., Hodgson, E. G., Atabani, S., De Souza, K. et al. Clinical impact of tumor DNA repair expression and T-cell infiltration in breast cancers. Cancer Immunol. Res. 5, 292–299 (2017).

  25. 25.

    McCarty, K. S. Jr. & KS, Mc. Carty Sr. Histochemical approaches to steroid receptor analyses. Semin. Diagn. Pathol. 1, 297–308 (1984).

  26. 26.

    Elston, C. W. & Ellis, I. O. Pathological prognostic factors in breast cancer. I. The value of histological grade in breast cancer: experience from a large study with long-term follow-up. C. W. Elston & I. O. Ellis. Histopathology 19, 403–410 (1991). Histopathology 41,151–152 (2002), discussion 2–3.

  27. 27.

    Senkus, E., Kyriakides, S., Ohno, S., Penault-Llorca, F., Poortmans, P., Rutgers, E. et al. Primary breast cancer: ESMO Clinical Practice Guidelines for diagnosis, treatment and follow-up. Ann. Oncol. 26(Suppl 5), v8–v30 (2015).

  28. 28.

    Perou, C. M., Sorlie, T., Eisen, M. B., van de Rijn, M., Jeffrey, S. S., Rees, C. A. et al. Molecular portraits of human breast tumours. Nature 406, 747–752 (2000).

  29. 29.

    Kim, S., Kim, D. H., Jung, W. H. & Koo, J. S. Expression of glutamine metabolism-related proteins according to molecular subtype of breast cancer. Endocr. Relat. Cancer 20, 339–348 (2013).

  30. 30.

    Hilvo, M., Denkert, C., Lehtinen, L., Muller, B., Brockmoller, S., Seppanen-Laakso, T. et al. Novel theranostic opportunities offered by characterization of altered membrane lipid metabolism in breast cancer progression. Cancer Res. 71, 3236–3245 (2011).

  31. 31.

    Gatenby, R. A. & Gillies, R. J. Why do cancers have high aerobic glycolysis? Nat. Rev. Cancer. 4, 891–899 (2004).

  32. 32.

    Netea-Maier, R. T., Smit, J. W. A. & Netea, M. G. Metabolic changes in tumor cells and tumor-associated macrophages: A mutual relationship. Cancer Lett. 413, 102–109 (2018).

  33. 33.

    Althobiti, M., Aleskandarany, M. A., Joseph, C., Toss, M., Mongan, N., Diez-Rodriguez, M. et al. Heterogeneity of tumour infiltrating lymphocytes (TILs) in breast cancer and its prognostic significance. Histopathology 73, 887–896 (2018).

  34. 34.

    Martinez-Outschoorn, U. E., Lin, Z., Ko, Y. H., Goldberg, A. F., Flomenberg, N., Wang, C. et al. Understanding the metabolic basis of drug resistance: therapeutic induction of the Warburg effect kills cancer cells. Cell Cycle 10, 2521–2528 (2011).

  35. 35.

    Zhang, W. J., Chen, C., Zhou, Z. H., Gao, S. T., Tee, T. J., Yang, L. Q. et al. Hypoxia-inducible factor-1 alpha correlates with tumor-associated macrophages infiltration, influences survival of gastric cancer patients. J. Cancer 8, 1818–1825 (2017).

  36. 36.

    Li, N., Li, Y., Li, Z., Huang, C., Yang, Y., Lang, M. et al. Hypoxia inducible factor 1 (HIF-1) recruits macrophage to activate pancreatic stellate cells in pancreatic ductal adenocarcinoma. Int. J. Mol. Sci. 17, 799–811 (2016).

  37. 37.

    Land, S. C. & Tee, A. R. Hypoxia-inducible factor 1alpha is regulated by the mammalian target of rapamycin (mTOR) via an mTOR signaling motif. J. Biol. Chem. 282, 20534–20543 (2007).

  38. 38.

    Mittendorf, E. A., Philips, A. V., Meric-Bernstam, F., Qiao, N., Wu, Y., Harrington, S. et al. PD-L1 expression in triple-negative breast cancer. Cancer Immunol. Res. 2, 361–370 (2014).

  39. 39.

    Tsang, J. Y., Au, W. L., Lo, K. Y., Ni, Y. B., Hlaing, T., Hu, J. et al. PD-L1 expression and tumor infiltrating PD-1+ lymphocytes associated with outcome in HER2+ breast cancer patients. Breast Cancer Res. Tr. 162, 19–30 (2017).

  40. 40.

    Rohde, T., MacLean, D. A. & Klarlund Pedersen, B. Glutamine, lymphocyte proliferation and cytokine production. Scand J. Immunol. 44, 648–650 (1996).

  41. 41.

    Nakaya, M., Xiao, Y., Zhou, X., Chang, J. H., Chang, M., Cheng, X. et al. Inflammatory T cell responses rely on amino acid transporter ASCT2 facilitation of glutamine uptake and mTORC1 kinase activation. Immunity. 40, 692–705 (2014).

  42. 42.

    Sinclair, L. V., Rolf, J., Emslie, E., Shi, Y. B., Taylor, P. M. & Cantrell, D. A. Control of amino-acid transport by antigen receptors coordinates the metabolic reprogramming essential for T cell differentiation. Nat. Immunol. 14, 500–508 (2013).

  43. 43.

    Ren, W., Liu, G., Yin, J., Tan, B., Wu, G., Bazer, F. W. et al. Amino-acid transporters in T-cell activation and differentiation. Cell Death Dis. 8, e2757 (2017).

  44. 44.

    Ligthart-Melis, G. C., van de Poll, M. C., Boelens, P. G., Dejong, C. H., Deutz, N. E. & van Leeuwen, P. A. Glutamine is an important precursor for de novo synthesis of arginine in humans. Am. J. Clin. Nutr. 87, 1282–1289 (2008).

  45. 45.

    Biswas, S. K. & Mantovani, A. Orchestration of metabolism by macrophages. Cell Metab. 15, 432–437 (2012).

  46. 46.

    Penkert, J., Ripperger, T., Schieck, M., Schlegelberger, B., Steinemann, D. & Illig, T. On metabolic reprogramming and tumor biology: A comprehensive survey of metabolism in breast cancer. Oncotarget 7, 67626 (2016).

  47. 47.

    Liu, Y., Yang, L., An, H., Chang, Y., Zhang, W., Zhu, Y. et al. High expression of solute carrier family 1, member 5 (SLC1A5) is associated with poor prognosis in clear-cell renal cell carcinoma. Sci. Rep. 5, 16954 (2015).

  48. 48.

    Muenst, S., Soysal, S. D., Gao, F., Obermann, E. C., Oertli, D. & Gillanders, W. E. The presence of programmed death 1 (PD-1)-positive tumor-infiltrating lymphocytes is associated with poor prognosis in human breast cancer. Breast Cancer Res. Tr. 139, 667–676 (2013).

  49. 49.

    Muenst, S., Schaerli, A. R., Gao, F., Daster, S., Trella, E., Droeser, R. A. et al. Expression of programmed death ligand 1 (PD-L1) is associated with poor prognosis in human breast cancer. Breast Cancer Res. Tr. 146, 15–24 (2014).

  50. 50.

    Baptista, M. Z., Sarian, L. O., Derchain, S. F., Pinto, G. A. & Vassallo, J. Prognostic significance of PD-L1 and PD-L2 in breast cancer. Hum. Pathol. 47, 78–84 (2016).

  51. 51.

    Parsa, A. T., Waldron, J. S., Panner, A., Crane, C. A., Parney, I. F., Barry, J. J. et al. Loss of tumor suppressor PTEN function increases B7-H1 expression and immunoresistance in glioma. Nat. Med. 13, 84–88 (2007).

  52. 52.

    Lastwika, K. J., Wilson, W. 3rd, Li, Q. K., Norris, J., Xu, H., Ghazarian, S. R. et al. Control of PD-L1 expression by oncogenic activation of the AKT-mTOR pathway in non-small cell lung cancer. Cancer Res. 76, 227–238 (2016).

  53. 53.

    Brahmer, J. R., Tykodi, S. S., Chow, L. Q., Hwu, W. J., Topalian, S. L., Hwu, P. et al. Safety and activity of anti-PD-L1 antibody in patients with advanced cancer. N. Engl. J. Med. 366, 2455–2465 (2012).

  54. 54.

    Topalian, S. L., Hodi, F. S., Brahmer, J. R., Gettinger, S. N., Smith, D. C., McDermott, D. F. et al. Safety, activity, and immune correlates of anti-PD-1 antibody in cancer. N. Engl. J. Med. 366, 2443–2454 (2012).

  55. 55.

    Powles, T., Eder, J. P., Fine, G. D., Braiteh, F. S., Loriot, Y., Cruz, C. et al. MPDL3280A (anti-PD-L1) treatment leads to clinical activity in metastatic bladder cancer. Nature 515, 558–562 (2014).

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We thank the Nottingham Health Science Biobank and Breast Cancer Now Tissue Bank for the provision of tissue samples. We thank the University of Nottingham (Nottingham Life Cycle 6 and Cancer Research Priority Area) for funding.

Author contributions

R.E. contributed to writing, IHC staining, scoring, data analysis and interpretation; M.L.C. contributed to writing and reviewing the paper; M.A. and L.A. contributed to analysis and reviewing the paper; I.O.E. and E.A.R. contributed to writing and reviewing the paper; A.R.G. contributed to study design, data analysis and interpretation, writing and reviewing the paper.

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Correspondence to Andrew R. Green.

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The authors declare no competing interests.

Ethics approval and consent to participate

This study was approved by the Nottingham Research Ethics Committee 2 under the title ‘Development of a molecular genetic classification of breast cancer’ and the North West–Greater Manchester Central Research Ethics Committee under the title ‘Nottingham Health Science Biobank (NHSB)’ reference number 15/NW/0685. The study was performed in accordance with the Declaration of Helsinki.


Sources of study funding are the University of Nottingham (Nottingham Life Cycle 6 and Cancer Research Priority Area).

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The data sets generated during this study are available from the corresponding author on reasonable request.

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Ansari, R.E., Craze, M.L., Althobiti, M. et al. Enhanced glutamine uptake influences composition of immune cell infiltrates in breast cancer. Br J Cancer 122, 94–101 (2020).

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