Cancer Immunology, Immunotherapy

, Volume 60, Issue 5, pp 649–658 | Cite as

Generation and characterization of the first inhibitory antibody targeting tumour-associated carbonic anhydrase XII

  • Christina Battke
  • Elisabeth Kremmer
  • Josef Mysliwietz
  • Gabor Gondi
  • Claudia Dumitru
  • Sven Brandau
  • Stephan Lang
  • Daniela Vullo
  • Claudiu Supuran
  • Reinhard Zeidler
Original article

Abstract

The carbonic anhydrases (CAs) constitute a family of almost ubiquitous enzymes of significant importance for many physiological and pathological processes. CAs reversely catalyse the conversion of CO2 + H2O to HCO3 and H+, thereby contributing to the regulation of intracellular pH. Above all, CAs are of key importance for cells that perform glycolysis that inevitably leads to the intracellular accumulation of lactate. CA XII is a plasma membrane-associated isoform of the enzyme, which is induced by hypoxia and oestrogen and, consequently, expressed at high levels on various types of cancer and, intriguingly, on cancer stem cells. The enzyme is directly involved in tumour progression, and its inhibition has an anti-tumour effect. Apart from its role in carcinogenesis, the enzyme contributes to various other diseases like glaucoma and arteriosclerotic plaques, among others. CA XII is therefore regarded as promising target for specific therapies. We have now generated the first monoclonal antibody (6A10) that binds to the catalytic domain of CA XII on vital tumour cells and inhibits CA XII enzyme activity at nanomolar concentrations and thus much more effective than acetazolamide. In vitro results demonstrate that inhibition of CA XII by 6A10 inhibits the growth of tumour cells in 3-dimensional structures. In conclusion, we generated the first specific and efficient biological inhibitor of tumour-associated CA XII. This antibody may serve as a valuable tool for in vivo diagnosis and adjuvant treatment of different types of cancer.

Keywords

Carbonic anhydrase Monoclonal antibody Cancer treatment Hypoxia Warburg effect 

Abbreviations

CAs

Carbonic anhydrases

Supplementary material

262_2011_980_MOESM1_ESM.pdf (593 kb)
Supplementary material 1 (PDF 593 kb)

References

  1. 1.
    Warburg O (1956) On respiratory impairment in cancer cells. Science 124:269–270PubMedGoogle Scholar
  2. 2.
    Stubbs M, McSheehy PM, Griffiths JR, Bashford CL (2000) Causes and consequences of tumour acidity and implications for treatment. Mol Med Today 6:15–19PubMedCrossRefGoogle Scholar
  3. 3.
    Oksala N, Levula M, Pelto-Huikko M, Kytomaki L, Soini JT, Salenius J, Kahonen M, Karhunen PJ, Laaksonen R, Parkkila S, Lehtimaki T (2010) Carbonic anhydrases II and XII are up-regulated in osteoclast-like cells in advanced human atherosclerotic plaques-Tampere Vascular Study. Ann Med 42:360–370PubMedCrossRefGoogle Scholar
  4. 4.
    Liao SY, Ivanov S, Ivanova A, Ghosh S, Cote MA, Keefe K, Coca-Prados M, Stanbridge EJ, Lerman MI (2003) Expression of cell surface transmembrane carbonic anhydrase genes CA9 and CA12 in the human eye: overexpression of CA12 (CA XII) in glaucoma. J Med Genet 40:257–261PubMedCrossRefGoogle Scholar
  5. 5.
    Supuran CT (2008) Carbonic anhydrases—an overview. Curr Pharm Des 14:603–614PubMedCrossRefGoogle Scholar
  6. 6.
    Alterio V, Hilvo M, Di Fiore A, Supuran CT, Pan P, Parkkila S, Scaloni A, Pastorek J, Pastorekova S, Pedone C, Scozzafava A, Monti SM, De Simone G (2009) Crystal structure of the catalytic domain of the tumor-associated human carbonic anhydrase IX. Proc Natl Acad Sci USA 106:16233–16238PubMedCrossRefGoogle Scholar
  7. 7.
    Whittington DA, Waheed A, Ulmasov B, Shah GN, Grubb JH, Sly WS, Christianson DW (2001) Crystal structure of the dimeric extracellular domain of human carbonic anhydrase XII, a bitopic membrane protein overexpressed in certain cancer tumor cells. Proc Natl Acad Sci USA 98:9545–9550PubMedCrossRefGoogle Scholar
  8. 8.
    Wykoff CC, Beasley N, Watson PH, Campo L, Chia SK, English R, Pastorek J, Sly WS, Ratcliffe P, Harris AL (2001) Expression of the hypoxia-inducible and tumor-associated carbonic anhydrases in ductal carcinoma in situ of the breast. Am J Pathol 158:1011–1019PubMedCrossRefGoogle Scholar
  9. 9.
    Hsieh MJ, Chen KS, Chiou HL, Hsieh YS (2010) Carbonic anhydrase XII promotes invasion and migration ability of MDA-MB-231 breast cancer cells through the p38 MAPK signaling pathway. Eur J Cell Biol 89:598–606PubMedCrossRefGoogle Scholar
  10. 10.
    Chiche J, Ilc K, Laferriere J, Trottier E, Dayan F, Mazure NM, Brahimi-Horn MC, Pouyssegur J (2009) Hypoxia-inducible carbonic anhydrase IX and XII promote tumor cell growth by counteracting acidosis through the regulation of the intracellular pH. Cancer Res 69:358–368PubMedCrossRefGoogle Scholar
  11. 11.
    Parkkila S, Rajaniemi H, Parkkila AK, Kivela J, Waheed A, Pastorekova S, Pastorek J, Sly WS (2000) Carbonic anhydrase inhibitor suppresses invasion of renal cancer cells in vitro. Proc Natl Acad Sci USA 97:2220–2224PubMedCrossRefGoogle Scholar
  12. 12.
    Svastova E, Zilka N, Zat’ovicova M, Gibadulinova A, Ciampor F, Pastorek J, Pastorekova S (2003) Carbonic anhydrase IX reduces E-cadherin-mediated adhesion of MDCK cells via interaction with beta-catenin. Exp Cell Res 290:332–345PubMedCrossRefGoogle Scholar
  13. 13.
    Tureci O, Sahin U, Vollmar E, Siemer S, Gottert E, Seitz G, Parkkila AK, Shah GN, Grubb JH, Pfreundschuh M, Sly WS (1998) Human carbonic anhydrase XII: cDNA cloning, expression, and chromosomal localization of a carbonic anhydrase gene that is overexpressed in some renal cell cancers. Proc Natl Acad Sci USA 95:7608–7613PubMedCrossRefGoogle Scholar
  14. 14.
    Karhumaa P, Parkkila S, Tureci O, Waheed A, Grubb JH, Shah G, Parkkila A, Kaunisto K, Tapanainen J, Sly WS, Rajaniemi H (2000) Identification of carbonic anhydrase XII as the membrane isozyme expressed in the normal human endometrial epithelium. Mol Hum Reprod 6:68–74PubMedCrossRefGoogle Scholar
  15. 15.
    Kivela A, Parkkila S, Saarnio J, Karttunen TJ, Kivela J, Parkkila AK, Waheed A, Sly WS, Grubb JH, Shah G, Tureci O, Rajaniemi H (2000) Expression of a novel transmembrane carbonic anhydrase isozyme XII in normal human gut and colorectal tumors. Am J Pathol 156:577–584PubMedCrossRefGoogle Scholar
  16. 16.
    Barnett DH, Sheng S, Charn TH, Waheed A, Sly WS, Lin CY, Liu ET, Katzenellenbogen BS (2008) Estrogen receptor regulation of carbonic anhydrase XII through a distal enhancer in breast cancer. Cancer Res 68:3505–3515PubMedCrossRefGoogle Scholar
  17. 17.
    Hynninen P, Vaskivuo L, Saarnio J, Haapasalo H, Kivela J, Pastorekova S, Pastorek J, Waheed A, Sly WS, Puistola U, Parkkila S (2006) Expression of transmembrane carbonic anhydrases IX and XII in ovarian tumours. Histopathology 49:594–602PubMedCrossRefGoogle Scholar
  18. 18.
    Ilie MI, Hofman V, Ortholan C, Ammadi RE, Bonnetaud C, Havet K, Venissac N, Mouroux J, Mazure NM, Pouyssegur J, Hofman P (2010) Overexpression of carbonic anhydrase XII in tissues from resectable non-small cell lung cancers is a biomarker of good prognosis. Int J Cancer 128:1614–1623Google Scholar
  19. 19.
    Ivanov S, Liao SY, Ivanova A, Danilkovitch-Miagkova A, Tarasova N, Weirich G, Merrill MJ, Proescholdt MA, Oldfield EH, Lee J, Zavada J, Waheed A, Sly W, Lerman MI, Stanbridge EJ (2001) Expression of hypoxia-inducible cell-surface transmembrane carbonic anhydrases in human cancer. Am J Pathol 158:905–919PubMedCrossRefGoogle Scholar
  20. 20.
    Leppilampi M, Saarnio J, Karttunen TJ, Kivela J, Pastorekova S, Pastorek J, Waheed A, Sly WS, Parkkila S (2003) Carbonic anhydrase isozymes IX and XII in gastric tumors. World J Gastroenterol 9:1398–1403PubMedGoogle Scholar
  21. 21.
    Parkkila S, Parkkila AK, Saarnio J, Kivela J, Karttunen TJ, Kaunisto K, Waheed A, Sly WS, Tureci O, Virtanen I, Rajaniemi H (2000) Expression of the membrane-associated carbonic anhydrase isozyme XII in the human kidney and renal tumors. J Histochem Cytochem 48:1601–1608PubMedCrossRefGoogle Scholar
  22. 22.
    Proescholdt MA, Mayer C, Kubitza M, Schubert T, Liao SY, Stanbridge EJ, Ivanov S, Oldfield EH, Brawanski A, Merrill MJ (2005) Expression of hypoxia-inducible carbonic anhydrases in brain tumors. Neuro Oncology 7:465–475PubMedCrossRefGoogle Scholar
  23. 23.
    Ulmasov B, Waheed A, Shah GN, Grubb JH, Sly WS, Tu C, Silverman DN (2000) Purification and kinetic analysis of recombinant CA XII, a membrane carbonic anhydrase overexpressed in certain cancers. Proc Natl Acad Sci USA 97:14212–14217PubMedCrossRefGoogle Scholar
  24. 24.
    Ashida S, Nishimori I, Tanimura M, Onishi S, Shuin T (2002) Effects of von Hippel-Lindau gene mutation and methylation status on expression of transmembrane carbonic anhydrases in renal cell carcinoma. J Cancer Res Clin Oncol 128:561–568PubMedCrossRefGoogle Scholar
  25. 25.
    Creighton CJ, Cordero KE, Larios JM, Miller RS, Johnson MD, Chinnaiyan AM, Lippman ME, Rae JM (2006) Genes regulated by estrogen in breast tumor cells in vitro are similarly regulated in vivo in tumor xenografts and human breast tumors. Genome Biol 7:R28PubMedCrossRefGoogle Scholar
  26. 26.
    Frasor J, Stossi F, Danes JM, Komm B, Lyttle CR, Katzenellenbogen BS (2004) Selective estrogen receptor modulators: discrimination of agonistic versus antagonistic activities by gene expression profiling in breast cancer cells. Cancer Res 64:1522–1533PubMedCrossRefGoogle Scholar
  27. 27.
    Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65:55–63PubMedCrossRefGoogle Scholar
  28. 28.
    Ibrahim SF, Diercks AH, Petersen TW, van den Engh G (2007) Kinetic analyses as a critical parameter in defining the side population (SP) phenotype. Exp Cell Res 313:1921–1926PubMedCrossRefGoogle Scholar
  29. 29.
    Innocenti A, Vullo D, Pastorek J, Scozzafava A, Pastorekova S, Nishimori I, Supuran CT (2007) Carbonic anhydrase inhibitors. Inhibition of transmembrane isozymes XII (cancer-associated) and XIV with anions. Bioorg Med Chem Lett 17:1532–1537PubMedCrossRefGoogle Scholar
  30. 30.
    Vullo D, Innocenti A, Nishimori I, Pastorek J, Scozzafava A, Pastorekova S, Supuran CT (2005) Carbonic anhydrase inhibitors. Inhibition of the transmembrane isozyme XII with sulfonamides-a new target for the design of antitumor and antiglaucoma drugs? Bioorg Med Chem Lett 15:963–969PubMedCrossRefGoogle Scholar
  31. 31.
    Khalifah RG (1971) The carbon dioxide hydration activity of carbonic anhydrase. I. Stop-flow kinetic studies on the native human isoenzymes B and C. J Biol Chem 246:2561–2573PubMedGoogle Scholar
  32. 32.
    Supuran CT (2008) Carbonic anhydrases: novel therapeutic applications for inhibitors and activators. Nat Rev Drug Discov 7:168–181PubMedCrossRefGoogle Scholar
  33. 33.
    Supuran CT, Scozzafava A, Casini A (2003) Carbonic anhydrase inhibitors. Med Res Rev 23:146–189PubMedCrossRefGoogle Scholar
  34. 34.
    Pastorekova S, Zatovicova M, Pastorek J (2008) Cancer-associated carbonic anhydrases and their inhibition. Curr Pharm Des 14:685–698PubMedCrossRefGoogle Scholar
  35. 35.
    Jensen RL (2009) Brain tumor hypoxia: tumorigenesis, angiogenesis, imaging, pseudoprogression, and as a therapeutic target. J Neurooncol 92:317–335PubMedCrossRefGoogle Scholar
  36. 36.
    Supuran CT, Scozzafava A (2007) Carbonic anhydrases as targets for medicinal chemistry. Bioorg Med Chem 15:4336–4350PubMedCrossRefGoogle Scholar
  37. 37.
    Swietach P, Wigfield S, Supuran CT, Harris AL, Vaughan-Jones RD (2008) Cancer-associated, hypoxia-inducible carbonic anhydrase IX facilitates CO2 diffusion. BJU Int 101(Suppl 4):22–24PubMedCrossRefGoogle Scholar
  38. 38.
    Swietach P, Wigfield S, Cobden P, Supuran CT, Harris AL, Vaughan-Jones RD (2008) Tumor-associated carbonic anhydrase 9 spatially coordinates intracellular pH in three-dimensional multicellular growths. J Biol Chem 283:20473–20483PubMedCrossRefGoogle Scholar
  39. 39.
    Deonarain MP, Kousparou CA, Epenetos AA (2009) Antibodies targeting cancer stem cells: a new paradigm in immunotherapy? MAbs 1:12–25PubMedCrossRefGoogle Scholar
  40. 40.
    Wu C, Alman BA (2008) Side population cells in human cancers. Cancer Lett 268:1–9PubMedCrossRefGoogle Scholar
  41. 41.
    Sung JM, Cho HJ, Yi H, Lee CH, Kim HS, Kim DK, Abd El-Aty AM, Kim JS, Landowski CP, Hediger MA, Shin HC (2008) Characterization of a stem cell population in lung cancer A549 cells. Biochem Biophys Res Commun 371:163–167PubMedCrossRefGoogle Scholar
  42. 42.
    Vaupel P, Mayer A (2007) Hypoxia in cancer: significance and impact on clinical outcome. Cancer Metastasis Rev 26:225–239PubMedCrossRefGoogle Scholar
  43. 43.
    Isa AY, Ward TH, West CM, Slevin NJ, Homer JJ (2006) Hypoxia in head and neck cancer. Br J Radiol 79:791–798PubMedCrossRefGoogle Scholar
  44. 44.
    Hui EP, Chan AT, Pezzella F, Turley H, To KF, Poon TC, Zee B, Mo F, Teo PM, Huang DP, Gatter KC, Johnson PJ, Harris 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 8:2595–2604PubMedGoogle Scholar
  45. 45.
    Kappler M, Taubert H, Holzhausen HJ, Reddemann R, Rot S, Becker A, Kuhnt T, Dellas K, Dunst J, Vordermark D, Hansgen G, Bache M (2008) Immunohistochemical detection of HIF-1alpha and CAIX in advanced head-and-neck cancer. Prognostic role and correlation with tumor markers and tumor oxygenation parameters. Strahlenther Onkol 184:393–399PubMedCrossRefGoogle Scholar
  46. 46.
    Beasley NJ, Wykoff CC, Watson PH, Leek R, Turley H, Gatter K, Pastorek J, Cox GJ, Ratcliffe P, Harris AL (2001) Carbonic anhydrase IX, an endogenous hypoxia marker, expression in head and neck squamous cell carcinoma and its relationship to hypoxia, necrosis, and microvessel density. Cancer Res 61:5262–5267PubMedGoogle Scholar
  47. 47.
    Haapasalo J, Hilvo M, Nordfors K, Haapasalo H, Parkkila S, Hyrskyluoto A, Rantala I, Waheed A, Sly WS, Pastorekova S, Pastorek J, Parkkila AK (2008) Identification of an alternatively spliced isoform of carbonic anhydrase XII in diffusely infiltrating astrocytic gliomas. Neuro Oncology 10:131–138PubMedCrossRefGoogle Scholar
  48. 48.
    Fang JS, Gillies RD, Gatenby RA (2008) Adaptation to hypoxia and acidosis in carcinogenesis and tumor progression. Semin Cancer Biol 18:330–337PubMedCrossRefGoogle Scholar
  49. 49.
    Ke Q, Costa M (2006) Hypoxia-inducible factor-1 (HIF-1). Mol Pharmacol 70:1469–1480PubMedCrossRefGoogle Scholar
  50. 50.
    Silva AS, Yunes JA, Gillies RJ, Gatenby RA (2009) The potential role of systemic buffers in reducing intratumoral extracellular pH and acid-mediated invasion. Cancer Res 69:2677–2684PubMedCrossRefGoogle Scholar
  51. 51.
    Oosterwijk E, Ruiter DJ, Hoedemaeker PJ, Pauwels EK, Jonas U, Zwartendijk J, Warnaar SO (1986) Monoclonal antibody G 250 recognizes a determinant present in renal-cell carcinoma and absent from normal kidney. Int J Cancer 38:489–494PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Christina Battke
    • 1
  • Elisabeth Kremmer
    • 2
  • Josef Mysliwietz
    • 2
  • Gabor Gondi
    • 1
  • Claudia Dumitru
    • 3
  • Sven Brandau
    • 3
  • Stephan Lang
    • 3
  • Daniela Vullo
    • 4
  • Claudiu Supuran
    • 4
  • Reinhard Zeidler
    • 5
  1. 1.Department of Gene VectorsHelmholtz-CenterMunichGermany
  2. 2.Institute of Molecular ImmunologyHelmholtz-CenterMunichGermany
  3. 3.ENT-DepartmentUniversitätsklinikum EssenEssenGermany
  4. 4.Laboratorio di Chimica BioinorganicaUniversità degli Studi di FirenzeSesto Fiorentino (Firenze)Italy
  5. 5.ENT-DepartmentLudwig-Maximilians-UniversityMunichGermany

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