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Short Communications

Inhibition of vacuolar ATPase subunit in tumor cells delays tumor growth by decreasing the essential macrophage population in the tumor microenvironment

Abstract

In cancer cells, vacuolar ATPase (V-ATPase), a multi-subunit enzyme, is expressed on the plasma as well as vesicular membranes and critically influences metastatic behavior. The soluble, cleaved N-terminal domain of V-ATPase a2 isoform is associated with in vitro induction of tumorigenic characteristics in macrophages. This activity led us to further investigate its in vivo role in cancer progression by inhibition of a2 isoform (a2V) in tumor cells and the concomitant effect on tumor microenvironment in the mouse 4T-1 breast cancer model. Results showed that macrophages cocultivated with a2V knockdown (sh-a2) 4T-1 cells produce lower amounts of tumorigenic factors in vitro and have reduced ability to suppress T-cell activation and proliferation compared with control 4T-1 cells. Data analysis showed a delayed mammary tumor growth in Balb/c mice inoculated with sh-a2 4T-1 cells compared with control. The purified CD11b+ macrophages from sh-a2 tumors showed a reduced expression of mannose receptor-1 (CD206), interleukin-10, transforming growth factor-β, arginase-1, matrix metalloproteinase and vascular endothelial growth factor. Flow cytometric analysis of tumor-infiltrated macrophages showed a significantly low number of F4/80+CD11c+CD206+ macrophages in sh-a2 tumors compared with control. In sh-a2 tumors, most of the macrophages were F4/80+CD11c+ (antitumor M1 macrophages) suggesting it to be the reason behind delayed tumor growth. Additionally, tumor-infiltrating macrophages from sh-a2 tumors showed a reduced expression of CD206 compared with control whereas CD11c expression was unaffected. These findings demonstrate that in the absence of a2V in tumor cells, the resident macrophage population in the tumor microenvironment is altered which affects in vivo tumor growth. We suggest that by involving the host immune system, tumor growth can be controlled through targeting of a2V on tumor cells.

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References

  1. Umemura N, Saio M, Suwa T, Kitoh Y, Bai J, Nonaka K et al. Tumor-infiltrating myeloid-derived suppressor cells are pleiotropic-inflamed monocytes/macrophages that bear M1- and M2-type characteristics. J Leukoc Biol 2008; 83: 1136–1144.

    Article  CAS  Google Scholar 

  2. Sica A, Bronte V . Altered macrophage differentiation and immune dysfunction in tumor development. J Clin Invest 2007; 117: 1155–1166.

    Article  CAS  Google Scholar 

  3. Gabrilovich DI, Ostrand-Rosenberg S, Bronte V . Coordinated regulation of myeloid cells by tumours. Nat Rev Immunol 2012; 12: 253–268.

    Article  CAS  Google Scholar 

  4. Motz GT, Coukos G . Deciphering and reversing tumor immune suppression. Immunity 2013; 39: 61–73.

    Article  CAS  Google Scholar 

  5. Conejo-Garcia JR, Benencia F, Courreges MC, Kang E, Mohamed-Hadley A, Buckanovich RJ et al. Tumor-infiltrating dendritic cell precursors recruited by a beta-defensin contribute to vasculogenesis under the influence of Vegf-A. Nat Med 2004; 10: 950–958.

    Article  CAS  Google Scholar 

  6. Roy N, Pollard JW . Tumor-Associated Macrophages: From Mechanisms to Therapy. Immunity 2014; 41: 49–61.

    Article  Google Scholar 

  7. Katara GK, Jaiswal MK, Kulshrestha A, Kolli B, Gilman-Sachs A, Beaman KD . Tumor-associated vacuolar ATPase subunit promotes tumorigenic characteristics in macrophages. Oncogene 2013; 33: 5649–5654.

    Article  Google Scholar 

  8. Sennoune SR, Luo D, Martínez-Zaguilán R . Plasmalemmal vacuolar-type H+-ATPase in cancer biology. Cell Biochem Biophys 2004; 40: 185–206.

    Article  CAS  Google Scholar 

  9. Torigoe T, Izumi H, Ishiguchi H, Uramoto H, Murakami T, Ise T et al. Enhanced expression of the human vacuolar Hþ-ATPase c subunit gene (ATP6L) in response to anticancer agents. J Biol Chem 2002; 277: 36534–36543.

    Article  CAS  Google Scholar 

  10. Sennoune SR, Bakunts K, Martínez GM, Chua-Tuan JL, Kebir Y, Attaya MN et al. Vacuolar H+-ATPase in human breast cancer cells with distinct metastatic potential: distribution and functional activity. Am J Physiol Cell Physiol 2004; 286: C1443–C1452.

    Article  CAS  Google Scholar 

  11. Kwong C, Gilman-Sachs A, Beaman K . Tumor-associated a2 vacuolar ATPase acts as a key mediator of cancer-related inflammation by inducing pro-tumorigenic properties in monocytes. J Immunol 2011; 186: 1781–1789.

    Article  CAS  Google Scholar 

  12. Ntrivalas E, Gilman-Sachs A, Kwak-Kim J, Beaman K . The Nterminus domain of the a2 isoform of vacuolar ATPase can regulate interleukin- 1b production from mononuclear cells in co-culture with JEG-3 choriocarcinoma cells. Am J Reprod Immunol 2007; 57: 201–209.

    Article  CAS  Google Scholar 

  13. Jaiswal MK, Mallers TM, Larsen B, Kwak-Kim J, Chaouat G, Gilman-Sachs A et al. V-ATPase upregulation during early pregnancy: a possible link to establishment of an inflammatory response during preimplantation period of pregnancy. Reproduction 2012; 143: 713–725.

    Article  CAS  Google Scholar 

  14. Jaiswal MK, Agrawal V, Mallers T, Gilman-Sachs A, Hirsch E, Beaman KD . Regulation of apoptosis and innate immune stimuli in inflammation-induced preterm labor. J Immunol 2013; 191: 5702–5713.

    Article  CAS  Google Scholar 

  15. Kwong C, Gilman-Sachs A, Beaman K . An independent endocytic pathway stimulates different monocyte subsets by the a2 N-terminus domain of vacuolar-ATPase. Oncoimmunology 2013; 2: e22978.

    Article  Google Scholar 

  16. Forgac M . Vacuolar ATPases: rotary proton pumps in physiology and pathophysiology. Nat Rev Mol Cell Biol 2007; 8: 917–929.

    Article  CAS  Google Scholar 

  17. Hinton A, Sennoune SR, Bond S, Fang M, Reuveni M, Sahagian GG et al. Function of a subunit isoforms of the V-ATPase in pH homeostasis and in vitro invasion of MDA-MB231 human breast cancer cells. J Biol Chem 2009; 284: 16400–16408.

    Article  CAS  Google Scholar 

  18. Nishisho T, Hata K, Nakanishi M, Morita Y, Sun-Wada GH, Wada Y et al. The a3 isoform vacuolar type H+-ATPase promotes distant metastasis in the mouse B16 melanoma cells. Mol Cancer Res 2011; 9: 845–855.

    Article  CAS  Google Scholar 

  19. Mantovani A, Sozzani S, Locati M, Allavena P, Sica A . Macrophage polarization: tumor-associated macrophages as a paradigm for polarized M2 mononuclear phagocytes. Trends Immunol 2002; 23: 549–555.

    Article  CAS  Google Scholar 

  20. Sica A, Schioppa T, Mantovani A, Allavena P . Tumour-associated macrophages are a distinct M2 polarised population promoting tumour progression: potential targets of anti-cancer therapy. Eur J Cancer 2006; 42: 717–727.

    Article  CAS  Google Scholar 

  21. Ostrand-Rosenberg S . Myeloid-derived suppressor cells: more mechanisms for inhibiting antitumor immunity. Cancer Immunol Immunother 2010; 59: 1593–1600.

    Article  Google Scholar 

  22. Movahedi K, Laoui D, Gysemans C, Baeten M, Stange G, Van den Bossche J et al. Different tumor microenvironments contain functionally distinct subsets of macrophages derived from Ly6C(high) monocytes. Cancer Res 2010; 70: 5728–5739.

    Article  CAS  Google Scholar 

  23. Feng S, Zhu G, McConnell M, Deng L, Zhao Q, Wu M et al. Silencing of Atp6v1c1 prevents breast cancer growth and bone metastasis. Int J Biol Sci 2013; 9: 853–862.

    Article  CAS  Google Scholar 

  24. Lu X, Qin W, Li J, Tan N, Pan D, Zhang H et al. The growth and metastasis of human hepatocellular carcinoma xenografts are inhibited by small interfering RNA targeting to the subunit ATP6L of proton pump. Cancer Res 2005; 65: 6843–6849.

    Article  CAS  Google Scholar 

  25. Martinez FO, Gordon S, Locati M, Manotvani A . Transcriptional profiling of the human monocyte-to macrophage differentiation and polarization: new molecules and patterns of gene expression. J Immunol 2006; 199: 7303–7311.

    Article  Google Scholar 

  26. Granata F, Frattini A, Loffredo S, Staiano RI, Petraroli A, Ribatti D et al. Production of vascular endothelial growth factors from human lung macrophages induced by group IIA and group X secreted phospholipases A2. J Immunol 2010; 184: 5232–5241.

    Article  CAS  Google Scholar 

  27. Luo Y, Zhou H, Krueger J, Kaplan C, Lee SH, Dolman C et al. Targeting tumor-associated macrophages as a novel strategy against breast cancer. J Clin Invest 2006; 116: 2132–2141.

    Article  CAS  Google Scholar 

  28. Watkins SK, Egilmez NK, Suttles J, Stout RD . IL-12 rapidly alter the functional profiles of tumor-associated and tumor-infiltrating macrophages in vitro and in vivo. J Immunol 2007; 178: 1357–1362.

    Article  CAS  Google Scholar 

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Acknowledgements

We thank Robert Dickinson for technical assistance and the Rosalind Franklin University of Medicine and Science Flow Cytometry Core Facility. This work was supported by the grants from the Clinical Immunology Laboratory, Rosalind Franklin University of Medicine and Science, North Chicago, IL, USA.

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Correspondence to K D Beaman.

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Katara, G., Kulshrestha, A., Jaiswal, M. et al. Inhibition of vacuolar ATPase subunit in tumor cells delays tumor growth by decreasing the essential macrophage population in the tumor microenvironment. Oncogene 35, 1058–1065 (2016). https://doi.org/10.1038/onc.2015.159

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