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Carbonic anhydrase IX inhibition affects viability of cancer cells adapted to extracellular acidosis

Journal of Molecular Medicine volume 95, pages 1341–1353 (2017)Cite this article

Abstract

Among the players of the adaptive response of cancer cells able to promote a resistant and aggressive phenotype, carbonic anhydrase IX (CAIX) recently has emerged as one of the most relevant drug targets. Indeed, CAIX targeting has received a lot of interest, and selective inhibitors are currently under clinical trials. Hypoxia has been identified as the master inductor of CAIX, but, to date, very few is known about the influence that another important characteristic of tumor microenvironment, i.e., extracellular acidosis, exerts on CAIX expression and activity. In the last decades, acidic microenvironment has been associated with aggressive tumor phenotype endowed with epithelial-to-mesenchymal transition (EMT) profile, high invasive and migratory ability, apoptosis, and drug resistance. We demonstrated that melanoma, breast, and colorectal cancer cells transiently and chronically exposed to acidified medium (pH 6.7 ± 0.1) showed a significantly increased CAIX expression compared to those grown in standard conditions (pH 7.4 ± 0.1). Moreover, we observed that the CAIX inhibitor FC16-670A (also named SLC-0111, which just successfully ended phase I clinical trials) not only prevents such increased expression under acidosis but also promotes apoptotic and necrotic programs only in acidified cancer cells. Thus, CAIX could represent a selective target of acidic cancer cells and FC16-670A inhibitor as a useful tool to affect this aggressive subpopulation characterized by conventional therapy escape.

Key messages

  • Cancer cells overexpress CAIX under transient and chronic extracellular acidosis.

  • Acidosis-induced CAIX overexpression is NF-κB mediated and HIF-1α independent.

  • FC16-670A prevents CAIX overexpression and induces acidified cancer cell death.

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References

  1. Peppicelli S, Andreucci E, Ruzzolini J, Margheri F, Laurenzana A, Bianchini F, Calorini L (2017) Acidity of microenvironment as a further driver of tumor metabolic reprogramming. J Clin Cell Immunol 8:485

    Article  Google Scholar 

  2. Damaghi M, Wojtkowiak JW, Gillies RJ (2013) pH sensing and regulation in cancer. Front Physiol 4:370

    Article  PubMed  PubMed Central  Google Scholar 

  3. Neri D, Supuran CT (2011) Interfering with pH regulation in tumours as a therapeutic strategy. Nat Rev Drug Discov 10(10):767–777

    CAS  Article  PubMed  Google Scholar 

  4. Tafreshi NK, Lloyd MC, Bui MM, Gillies RJ, Morse DL (2014) Carbonic anhydrase IX as an imaging and therapeutic target for tumors and metastases. Subcell Biochem 75:221–254

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  5. Thiry A, Dogné JM, Masereel B, Supuran CT (2006) Targeting tumor-associated carbonic anhydrase IX in cancer therapy. Trends Pharmacol Sci 27(11):566–573

    CAS  Article  PubMed  Google Scholar 

  6. Zatovicova M, Sedlakova O, Svastova E, Ohradanova A, Ciampor F, Arribas J, Pastorek J, Pastorekova S (2005) Ectodomain shedding of the hypoxia-induced carbonic anhydrase IX is a metalloprotease-dependent process regulated by TACE/ADAM17. Br J Cancer 93(11):1267–1276

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  7. Shin HJ, Rho SB, Jung DC, Han IO, Oh ES, Kim JY (2011) Carbonic anhydrase IX (CA9) modulates tumor-associated cell migration and invasion. J Cell Sci 124(Pt 7):1077–1087

    CAS  Article  PubMed  Google Scholar 

  8. Robertson N, Potter C, Harris AL (2004) Role of carbonic anhydrase IX in human tumor cell growth, survival, and invasion. Cancer Res 64(17):6160–6165

    CAS  Article  PubMed  Google Scholar 

  9. Svastová E, Zilka N, Zat’ovicová M, Gibadulinová A, Ciampor F, Pastorek J, Pastoreková S (2003) Carbonic anhydrase IX reduces E-cadherin-mediated adhesion of MDCK cells via interaction with beta-catenin. Exp Cell Res 290(2):332–345

    Article  PubMed  Google Scholar 

  10. Ivanov S, Liao SY, Ivanova A, Danilkovitch-Miagkova A, Tarasova N, Weirich G, Merrill MJ, Proescholdt MA, Oldfield EH, Lee J et al (2001) Expression of hypoxia-inducible cell-surface transmembrane carbonic anhydrases in human cancer. Am J Pathol 158(3):905–919

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  11. Giatromanolaki A, Koukourakis MI, Sivridis E, Pastorek J, Wykoff CC, Gatter KC, Harris AL (2001) Expression of hypoxia-inducible carbonic anhydrase-9 relates to angiogenic pathways and independently to poor outcome in non-small cell lung cancer. Cancer Res 61(21):7992–7998

    CAS  PubMed  Google Scholar 

  12. Wykoff CC, Beasley NJ, Watson PH, Turner KJ, Pastorek J, Sibtain A, Wilson GD, Turley H, Talks KL, Maxwell PH et al (2000) Hypoxia-inducible expression of tumor-associated carbonic anhydrases. Cancer Res 60(24):7075–7083

    CAS  PubMed  Google Scholar 

  13. De Simone G, Supuran CT (2010) Carbonic anhydrase IX: biochemical and crystallographic characterization of a novel antitumor target. Biochim Biophys Acta 1804(2):404–409

    Article  PubMed  Google Scholar 

  14. Hashim AI, Zhang X, Wojtkowiak JW, Martinez GV, Gillies RJ (2011) Imaging pH and metastasis. NMR Biomed 24(6):582–591

    PubMed  PubMed Central  Google Scholar 

  15. Fukumura D, Xu L, Chen Y, Gohongi T, Seed B, Jain RK (2001) Hypoxia and acidosis independently up-regulate vascular endothelial growth factor transcription in brain tumors in vivo. Cancer Res 61(16):6020–6024

    CAS  PubMed  Google Scholar 

  16. Chiche J, Brahimi-Horn MC, Pouysségur J (2010) Tumour hypoxia induces a metabolic shift causing acidosis: a common feature in cancer. J Cell Mol Med 14(4):771–794

    CAS  Article  PubMed  Google Scholar 

  17. Sørensen BS, Hao J, Overgaard J, Vorum H, Honoré B, Alsner J, Horsman MR (2005) Influence of oxygen concentration and pH on expression of hypoxia induced genes. Radiother Oncol 76(2):187–193

    Article  PubMed  Google Scholar 

  18. Sørensen BS, Alsner J, Overgaard J, Horsman MR (2007) Hypoxia induced expression of endogenous markers in vitro is highly influenced by pH. Radiother Oncol 83(3):362–366

    Article  PubMed  Google Scholar 

  19. Tang X, Lucas JE, Chen JL, LaMonte G, Wu J, Wang MC, Koumenis C, Chi JT (2012) Functional interaction between responses to lactic acidosis and hypoxia regulates genomic transcriptional outputs. Cancer Res 72(2):491–502

    CAS  Article  PubMed  Google Scholar 

  20. Ihnatko R, Kubes M, Takacova M, Sedlakova O, Sedlak J, Pastorek J, Kopacek J, Pastorekova S (2006) Extracellular acidosis elevates carbonic anhydrase IX in human glioblastoma cells via transcriptional modulation that does not depend on hypoxia. Int J Oncol 29(4):1025–1033

    CAS  PubMed  Google Scholar 

  21. Peppicelli S, Bianchini F, Calorini L (2015) Dynamic scenario of metabolic pathway adaptation in tumors and therapeutic approach. Oncoscience 2(3):225–232

    Article  PubMed  PubMed Central  Google Scholar 

  22. Peppicelli S, Toti A, Giannoni E, Bianchini F, Margheri F, Del Rosso M, Calorini L (2016) Metformin is also effective on lactic acidosis-exposed melanoma cells switched to oxidative phosphorylation. Cell Cycle 15(14):1908–1918

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  23. Matsubara T, Diresta GR, Kakunaga S, Li D, Healey JH (2013) Additive influence of extracellular pH, oxygen tension, and pressure on invasiveness and survival of human osteosarcoma cells. Front Oncol 3:199

    Article  PubMed  PubMed Central  Google Scholar 

  24. Pacchiano F, Carta F, McDonald PC, Lou Y, Vullo D, Scozzafava A, Dedhar S, Supuran CT (2011) Ureido-substituted benzenesulfonamides potently inhibit carbonic anhydrase IX and show antimetastatic activity in a model of breast cancer metastasis. J Med Chem 54(6):1896–1902

    CAS  Article  PubMed  Google Scholar 

  25. Lou Y, McDonald PC, Oloumi A, Chia S, Ostlund C, Ahmadi A, Kyle A, Auf dem Keller U, Leung S, Huntsman D et al (2011) Targeting tumor hypoxia: suppression of breast tumor growth and metastasis by novel carbonic anhydrase IX inhibitors. Cancer Res 71(9):3364–3376

    CAS  Article  PubMed  Google Scholar 

  26. Carta F, Vullo D, Osman SM, AlOthman Z, Supuran CT (2017) Synthesis and carbonic anhydrase inhibition of a series of SLC-0111 analogs. Bioorg Med Chem 25(9):2569–2576

    CAS  Article  PubMed  Google Scholar 

  27. McCarty MF, Whitaker J (2010) Manipulating tumor acidification as a cancer treatment strategy. Altern Med Rev 15(3):264–272

    PubMed  Google Scholar 

  28. Webb BA, Chimenti M, Jacobson MP, Barber DL (2011) Dysregulated pH: a perfect storm for cancer progression. Nat Rev Cancer 11(9):671–677

    CAS  Article  PubMed  Google Scholar 

  29. Peppicelli S, Bianchini F, Calorini L (2014) Extracellular acidity, a “reappreciated” trait of tumor environment driving malignancy: perspectives in diagnosis and therapy. Cancer Metastasis Rev 33(2–3):823–832

    CAS  Article  PubMed  Google Scholar 

  30. Dubois L, Peeters S, Lieuwes NG, Geusens N, Thiry A, Wigfield S, Carta F, McIntyre A, Scozzafava A, Dogné JM et al (2011) Specific inhibition of carbonic anhydrase IX activity enhances the in vivo therapeutic effect of tumor irradiation. Radiother Oncol 99(3):424–431

    CAS  Article  PubMed  Google Scholar 

  31. Dubois L, Peeters SG, van Kuijk SJ, Yaromina A, Lieuwes NG, Saraya R, Biemans R, Rami M, Parvathaneni NK, Vullo D et al (2013) Targeting carbonic anhydrase IX by nitroimidazole based sulfamides enhances the therapeutic effect of tumor irradiation: a new concept of dual targeting drugs. Radiother Oncol 108(3):523–528

    CAS  Article  PubMed  Google Scholar 

  32. Sedlakova O, Svastova E, Takacova M, Kopacek J, Pastorek J, Pastorekova S (2014) Carbonic anhydrase IX, a hypoxia-induced catalytic component of the pH regulating machinery in tumors. Front Physiol 4:400

    Article  PubMed  PubMed Central  Google Scholar 

  33. Radvak P, Repic M, Svastova E, Takacova M, Csaderova L, Strnad H, Pastorek J, Pastorekova S, Kopacek J (2013) Suppression of carbonic anhydrase IX leads to aberrant focal adhesion and decreased invasion of tumor cells. Oncol Rep 29(3):1147–1153

    CAS  Article  PubMed  Google Scholar 

  34. Rofstad EK, Mathiesen B, Kindem K, Galappathi K (2006) Acidic extracellular pH promotes experimental metastasis of human melanoma cells in athymic nude mice. Cancer Res 66(13):6699–6707

    CAS  Article  PubMed  Google Scholar 

  35. Peppicelli S, Bianchini F, Torre E, Calorini L (2014) Contribution of acidic melanoma cells undergoing epithelial-to-mesenchymal transition to aggressiveness of non-acidic melanoma cells. Clin Exp Metastasis 31(4):423–433

    CAS  Article  PubMed  Google Scholar 

  36. Peppicelli S, Bianchini F, Calorini L (2015) Metabolic reprogramming as a continuous changing behavior of tumor cells. Tumour Biol 36(8):5759–5762

    CAS  Article  PubMed  Google Scholar 

  37. Wu H, Ying M, Hu X (2016) Lactic acidosis switches cancer cells from aerobic glycolysis back to dominant oxidative phosphorylation. Oncotarget 7(26):40621–40629

    PubMed  PubMed Central  Google Scholar 

  38. Hui EP, Chan AT, Pezzella F, Turley H, To KF, Poon TC, Zee B, Mo F, Teo PM, Huang DP et al (2002) Coexpression of hypoxia-inducible factors 1alpha and 2alpha, carbonic anhydrase IX, and vascular endothelial growth factor in nasopharyngeal carcinoma and relationship to survival. Clin Cancer Res 8(8):2595–2604

    CAS  PubMed  Google Scholar 

  39. Simko V, Takacova M, Debreova M, Laposova K, Ondriskova-Panisova E, Pastorekova S, Csaderova L, Pastorek J (2016) Dexamethasone downregulates expression of carbonic anhydrase IX via HIF-1α and NF-κB-dependent mechanisms. Int J Oncol 49(4):1277–1288

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  40. Peppicelli S, Andreucci E, Ruzzolini J, Laurenzana A, Margheri F, Fibbi G, Del Rosso M, Bianchini F, Calorini L (2017) The acidic microenvironment as a possible niche of dormant tumor cells. Cell Mol Life Sci 74(15):2761–2771

    CAS  Article  PubMed  Google Scholar 

  41. Svastová E, Hulíková A, Rafajová M, Zat’ovicová M, Gibadulinová A, Casini A, Cecchi A, Scozzafava A, Supuran CT, Pastorek J et al (2004) Hypoxia activates the capacity of tumor-associated carbonic anhydrase IX to acidify extracellular pH. FEBS Lett 577(3):439–445

    Article  PubMed  Google Scholar 

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Acknowledgements

This study was financially supported by grants from Istituto Toscano Tumori.

Author information

Affiliations

  1. Department of Clinical and Experimental Biomedical Sciences “Mario Serio”, University of Florence, Florence, Italy

    Elena Andreucci, Silvia Peppicelli, Jessica Ruzzolini, Francesca Bianchini & Lido Calorini

  2. Istituto Toscano Tumori (ITT), Florence, Italy

    Elena Andreucci, Silvia Peppicelli, Francesca Bianchini & Lido Calorini

  3. Department of NEUROFARBA, University of Florence, Florence, Italy

    Fabrizio Carta & Claudiu T. Supuran

  4. DISCOG, University of Padova, Padova, Italy

    Giulia Brisotto

  5. Immunology and Molecular Oncology Unit, IOV-IRCCS, Padova, Italy

    Giulia Brisotto

  6. Immunopathology and Cancer Biomarkers, Traslational Research Department, IRCCS, C.R.O. National Cancer Institute, Aviano, Pordenone, Italy

    Giulia Brisotto & Eva Biscontin

  7. Department of Surgery and Translational Medicine, University of Florence, Florence, Italy

    Alessio Biagioni

  8. Excellence Centre for Research, Transfer and High Education DENOTHE, Florence, Italy

    Lido Calorini

Authors
  1. Elena Andreucci
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  2. Silvia Peppicelli
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  3. Fabrizio Carta
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  5. Eva Biscontin
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  6. Jessica Ruzzolini
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  7. Francesca Bianchini
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  8. Alessio Biagioni
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  9. Claudiu T. Supuran
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  10. Lido Calorini
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Correspondence to Lido Calorini.

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Andreucci, E., Peppicelli, S., Carta, F. et al. Carbonic anhydrase IX inhibition affects viability of cancer cells adapted to extracellular acidosis. J Mol Med 95, 1341–1353 (2017). https://doi.org/10.1007/s00109-017-1590-9

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  • DOI: https://doi.org/10.1007/s00109-017-1590-9

Keywords

  • Acidosis of tumor microenvironment
  • Carbonic anhydrase IX (CAIX)
  • FC16-670A CAIX inhibitor
  • Apoptosis