Glutamine is an abundant and versatile nutrient in cancer cells. Head and neck squamous cell carcinoma (HNSCC) was reported to be dependent on mainly glucose, not glutamine, for producing the energy required for survival and proliferation.
The roles of ASCT2 (SLC1A5) and associated glutamine metabolism were determined by the MTT, colony formation, glutamine uptake, intracellular glutathione, ROS detection, immunofluorescence, immunohistochemistry, and apoptosis enzyme-linked immunosorbent assays as well as animal studies.
We found that glutamine is also critical for HNSCC. In this study, ASCT2, an amino acid transporter responsible for glutamine transport, in addition to LAT1 and GLS, is overexpressed in HNSCC and associated with poor survival. Using both in vivo and in vitro models, we found that knocking down ASCT2 by shRNAs or miR-137 or the combination of silencing ASCT2 and pharmacologically inhibiting SNAT2 via a small-molecule antagonist called V-9302 significantly suppressed intracellular glutamine levels and downstream glutamine metabolism, including glutathione production; these effects attenuated growth and proliferation, increased apoptosis and autophagy, and increased oxidative stress and mTORC1 pathway suppression in HNSCC. Additionally, silencing ASCT2 improved the response to cetuximab in HNSCC.
In summary, ASCT2-dependent glutamine uptake and subsequent glutamine metabolism are essential for HNSCC tumorigenesis, and the combination of glutamine uptake inhibitors and cetuximab presents a promising strategy for improving the outcomes of HNSCC patients.
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Vander Heiden, M. G. & DeBerardinis, R. J. Understanding the Intersections between Metabolism and Cancer Biology. Cell 168, 657–669 (2017).
DeBerardinis, R. J. & Chandel, N. S. Fundamentals of cancer metabolism. Sci. Adv. 2, e1600200 (2016).
Altman, B. J., Stine, Z. E. & Dang, C. V. From Krebs to clinic: glutamine metabolism to cancer therapy. Nat. Rev. Cancer 16, 619–634 (2016).
Hensley, C. T., Wasti, A. T. & DeBerardinis, R. J. Glutamine and cancer: cell biology, physiology, and clinical opportunities. J. Clin. Investig. 123, 3678–3684 (2013).
Villar, V. H., Merhi, F., Djavaheri-Mergny, M. & Duran, R. V. Glutaminolysis and autophagy in cancer. Autophagy 11, 1198–1208 (2015).
Kekuda, R., Prasad, P. D., Fei, Y. J., Torres-Zamorano, V., Sinha, S., Yang-Feng, T. L. et al. Cloning of the sodium-dependent, broad-scope, neutral amino acid transporter Bo from a human placental choriocarcinoma cell line. J. Biol. Chem. 271, 18657–18661 (1996).
Wang, Q., Hardie, R. A., Hoy, A. J., van Geldermalsen, M., Gao, D., Fazli, L. et al. Targeting ASCT2-mediated glutamine uptake blocks prostate cancer growth and tumour development. J. Pathol. 236, 278–289 (2015).
van Geldermalsen, M., Wang, Q., Nagarajah, R., Marshall, A. D., Thoeng, A., Gao, D. et al. ASCT2/SLC1A5 controls glutamine uptake and tumour growth in triple-negative basal-like breast cancer. Oncogene 35, 3201–3208 (2016).
Ye, J., Huang, Q., Xu, J., Huang, J., Wang, J., Zhong, W. et al. Targeting of glutamine transporter ASCT2 and glutamine synthetase suppresses gastric cancer cell growth. J. cancer Res. Clin. Oncol. 144, 821–833 (2018).
Liu, Y., Zhao, T., Li, Z., Wang, L., Yuan, S. & Sun, L. The role of ASCT2 in cancer: a review. Eur. J. Pharmacol. 837, 81–87 (2018).
Hassanein, M., Hoeksema, M. D., Shiota, M., Qian, J., Harris, B. K., Chen, H. et al. SLC1A5 mediates glutamine transport required for lung cancer cell growth and survival. Clin. Cancer Res. 19, 560–570 (2013).
Cormerais, Y., Massard, P. A., Vucetic, M., Giuliano, S., Tambutte, E., Durivault, J. et al. The glutamine transporter ASCT2 (SLC1A5) promotes tumor growth independently of the amino acid transporter LAT1 (SLC7A5). J. Biol. Chem. 293, 2877–2887 (2018).
Kamarajan, P., Rajendiran, T. M., Kinchen, J., Bermudez, M., Danciu, T. & Kapila, Y. L. Head and neck squamous cell carcinoma metabolism draws on glutaminolysis, and stemness is specifically regulated by glutaminolysis via aldehyde dehydrogenase. J. Proteome Res. 16, 1315–1326 (2017).
Yang, J., Guo, Y., Seo, W., Zhang, R., Lu, C., Wang, Y. et al. Targeting cellular metabolism to reduce head and neck cancer growth. Sci. Rep. 9, 4995 (2019).
Toyoda, M., Kaira, K., Ohshima, Y., Ishioka, N. S., Shino, M., Sakakura, K. et al. Prognostic significance of amino-acid transporter expression (LAT1, ASCT2, and xCT) in surgically resected tongue cancer. Br. J. Cancer 110, 2506–2513 (2014).
Schulte, M. L., Fu, A., Zhao, P., Li, J., Geng, L., Smith, S. T. et al. Pharmacological blockade of ASCT2-dependent glutamine transport leads to antitumor efficacy in preclinical models. Nat. Med. 24, 194–202 (2018).
Broer, A., Fairweather, S. & Broer, S. Disruption of amino acid homeostasis by novel ASCT2 inhibitors involves multiple targets. Front. Pharmacol. 9, 785 (2018).
Zhang, J., Pavlova, N. N. & Thompson, C. B. Cancer cell metabolism: the essential role of the nonessential amino acid, glutamine. EMBO J. 36, 1302–1315 (2017).
Broer, A., Rahimi, F. & Broer, S. Deletion of amino acid transporter ASCT2 (SLC1A5) reveals an essential role for transporters SNAT1 (SLC38A1) and SNAT2 (SLC38A2) to sustain glutaminolysis in cancer cells. J. Biol. Chem. 291, 13194–13205 (2016).
Tao, X., Lu, Y., Qiu, S., Wang, Y., Qin, J. & Fan, Z. AP1G1 is involved in cetuximab-mediated downregulation of ASCT2-EGFR complex and sensitization of human head and neck squamous cell carcinoma cells to ROS-induced apoptosis. Cancer Lett. 408, 33–42 (2017).
Luo, M., Wu, L., Zhang, K., Wang, H., Zhang, T., Gutierrez, L. et al. miR-137 regulates ferroptosis by targeting glutamine transporter SLC1A5 in melanoma. Cell Death Differ. 25, 1457–1472 (2018).
Lu, H., Li, X., Lu, Y., Qiu, S. & Fan, Z. ASCT2 (SLC1A5) is an EGFR-associated protein that can be co-targeted by cetuximab to sensitize cancer cells to ROS-induced apoptosis. Cancer Lett. 381, 23–30 (2016).
Moreadith, R. W. & Lehninger, A. L. The pathways of glutamate and glutamine oxidation by tumor cell mitochondria. Role of mitochondrial NAD(P)+-dependent malic enzyme. J. Biol. Chem. 259, 6215–6221 (1984).
Dong, J., Xiao, D., Zhao, Z., Ren, P., Li, C., Hu, Y. et al. Epigenetic silencing of microRNA-137 enhances ASCT2 expression and tumor glutamine metabolism. Oncogenesis 6, e356 (2017).
Jewell, J. L., Kim, Y. C., Russell, R. C., Yu, F. X., Park, H. W., Plouffe, S. W. et al. Metabolism. Differential regulation of mTORC1 by leucine and glutamine. Science 347, 194–198 (2015).
Bonnet, S., Archer, S. L., Allalunis-Turner, J., Haromy, A., Beaulieu, C., Thompson, R. et al. A mitochondria-K+ channel axis is suppressed in cancer and its normalization promotes apoptosis and inhibits cancer growth. Cancer Cell. 11, 37–51 (2007).
Graham, J., Muhsin, M. & Kirkpatrick, P. Cetuximab. Nat. Rev. Drug Discov. 3, 549–550 (2004).
Vander Heiden, M. G., Cantley, L. C. & Thompson, C. B. Understanding the Warburg effect: the metabolic requirements of cell proliferation. Science 324, 1029–1033 (2009).
Sandulache, V. C., Ow, T. J., Pickering, C. R., Frederick, M. J., Zhou, G., Fokt, I. et al. Glucose, not glutamine, is the dominant energy source required for proliferation and survival of head and neck squamous carcinoma cells. Cancer 117, 2926–2938 (2011).
Wang, J. B., Erickson, J. W., Fuji, R., Ramachandran, S., Gao, P., Dinavahi, R. et al. Targeting mitochondrial glutaminase activity inhibits oncogenic transformation. Cancer cell. 18, 207–19 (2010).
Cheng, T., Sudderth, J., Yang, C., Mullen, A. R., Jin, E. S., Mates, J. M. et al. Pyruvate carboxylase is required for glutamine-independent growth of tumor cells. Proc. Natl Acad. Sci. USA 108, 8674–8679 (2011).
Suzuki, S., Tanaka, T., Poyurovsky, M. V., Nagano, H., Mayama, T., Ohkubo, S. et al. Phosphate-activated glutaminase (GLS2), a p53-inducible regulator of glutamine metabolism and reactive oxygen species. Proc. Natl Acad. Sci. USA 107, 7461–7466 (2010).
Leemans, C. R., Snijders, P. J. F. & Brakenhoff, R. H. The molecular landscape of head and neck cancer. Nat. Rev. Cancer 18, 269–282 (2018).
Fleming, J. C., Woo, J., Moutasim, K., Mellone, M., Frampton, S. J., Mead, A. et al. HPV, tumour metabolism and novel target identification in head and neck squamous cell carcinoma. Br. J. Cancer 120, 356–367 (2019).
Dejure, F. R. & Eilers, M. MYC and tumor metabolism: chicken and egg. EMBO J. 36, 3409–3420 (2017).
Stine, Z. E., Walton, Z. E., Altman, B. J., Hsieh, A. L. & Dang, C. V. MYC, Metabolism, and cancer. Cancer Discov. 5, 1024–1039 (2015).
Gao, P., Tchernyshyov, I., Chang, T. C., Lee, Y. S., Kita, K., Ochi, T. et al. c-Myc suppression of miR-23a/b enhances mitochondrial glutaminase expression and glutamine metabolism. Nature 458, 762–765 (2009).
Langevin, S. M., Stone, R. A., Bunker, C. H., Lyons-Weiler, M. A., LaFramboise, W. A., Kelly, L. et al. MicroRNA-137 promoter methylation is associated with poorer overall survival in patients with squamous cell carcinoma of the head and neck. Cancer 117, 1454–1462 (2011).
Sullivan, L. B., Gui, D. Y., Hosios, A. M., Bush, L. N., Freinkman, E. & Vander Heiden, M. G. Supporting aspartate biosynthesis is an essential function of respiration in proliferating cells. Cell 162, 552–563 (2015).
DeBerardinis, R. J., Mancuso, A., Daikhin, E., Nissim, I., Yudkoff, M., Wehrli, S. et al. Beyond aerobic glycolysis: transformed cells can engage in glutamine metabolism that exceeds the requirement for protein and nucleotide synthesis. Proc. Natl Acad. Sci. USA 104, 19345–50 (2007).
Cosway, B. & Lovat, P. The role of autophagy in squamous cell carcinoma of the head and neck. Oral. Oncol. 54, 1–6 (2016).
Zhang, L., Zhang, W., Wang, Y. F., Liu, B., Zhang, W. F., Zhao, Y. F. et al. Dual induction of apoptotic and autophagic cell death by targeting survivin in head neck squamous cell carcinoma. Cell Death Dis. 6, e1771 (2015).
Huang, A. C., Lien, J. C., Lin, M. W., Yang, J. S., Wu, P. P., Chang, S. J. et al. Tetrandrine induces cell death in SAS human oral cancer cells through caspase activation-dependent apoptosis and LC3-I and LC3-II activation-dependent autophagy. Int. J. Oncol. 43, 485–94 (2013).
Chiu, M., Sabino, C., Taurino, G., Bianchi, M. G., Andreoli, R., Giuliani, N. et al. GPNA inhibits the sodium-independent transport system L for neutral amino acids. Amino Acids 49, 1365–1372 (2017).
Esslinger, C. S., Cybulski, K. A. & Rhoderick, J. F. Ngamma-aryl glutamine analogues as probes of the ASCT2 neutral amino acid transporter binding site. Bioorganic &. medicinal Chem. 13, 1111–1118 (2005).
Sacco, A. G. & Cohen, E. E. Current treatment options for recurrent or metastatic head and neck squamous cell carcinoma. J. Clin. Oncol. 33, 3305–3313 (2015).
Gyawali, B., Shimokata, T., Honda, K. & Ando, Y. Chemotherapy in locally advanced head and neck squamous cell carcinoma. Cancer Treat. Rev. 44, 10–16 (2016).
Ma, H., Wu, Z., Peng, J., Li, Y., Huang, H., Liao, Y. et al. Inhibition of SLC1A5 sensitizes colorectal cancer to cetuximab. Int. J. Cancer 142, 2578–2588 (2018).
Luo, J., Hong, Y., Lu, Y., Qiu, S., Chaganty, B. K., Zhang, L. et al. Acetyl-CoA carboxylase rewires cancer metabolism to allow cancer cells to survive inhibition of the Warburg effect by cetuximab. Cancer Lett. 384, 39–49 (2017).
We thank the TCGA project for its valuable contributions to HNSCC research.
The authors declare no competing interests.
Ethics approval and consent to participate
The study was conducted with approval from the ethics committee of Tianjin Cancer Hospital, and informed consent was obtained for experimentation. This study was performed in accordance to the Declaration of Helsinki.
This study was supported by the Natural Science Foundation of China (no. 81702698 and no. 81902757) and a grant from the Starting Foundation of MD/PhD and Introduced Talent issued by Tianjin Medical University Cancer Institute and Hospital. (no. B1602, B1720 and B1716).
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The datasets generated and analysed during the current study are available from the corresponding author on reasonable request.
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Zhang, Z., Liu, R., Shuai, Y. et al. ASCT2 (SLC1A5)-dependent glutamine uptake is involved in the progression of head and neck squamous cell carcinoma. Br J Cancer 122, 82–93 (2020). https://doi.org/10.1038/s41416-019-0637-9
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