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Epithelial tight junction proteins as potential antibody targets for pancarcinoma therapy

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Abstract

Recombinant monoclonal antibodies are beginning to revolutionize cancer therapy. In combination with standard chemotherapy, high response rates have been reported with antibodies of the human IgG1 isotype for treatment of non-Hodgkin’s lymphoma and breast cancer. It is becoming apparent that targets for antibody-based therapies do not necessarily need to be absent from normal tissues but can be present there either in low copy numbers or with binding epitopes shielded from the therapeutic antibody. Here, we studied whether claudin proteins that form tight junctions in normal epithelia are still expressed on carcinoma cells and whether their extracellular domains can be recognized by antibodies. We show that mRNAs of claudins 1, 3, 4, and 7 are all expressed in different human carcinoma cell lines, while claudin 8 was selectively expressed in breast and pancreas cancer lines. Chicken polyclonal antibodies were raised against peptides contained within predicted extracellular domains of claudins 1, 3, and 4. Affinity-purified IgG fractions for claudins 3 and 4 were monospecific and bound to human breast and colon carcinoma lines, but not to a line of monocytic origin. Claudin 3 antibodies also homogeneously stained human renal cell carcinoma tissue and micrometastatic tumor cells as identified by cytokeratin staining in bone marrow biopsies of breast cancer patients. Fluorescence-activated cell sorting and immunocytochemistry indicated that claudin antibodies bound to the surface of tumor cells. By analogy to other tumor-associated antigens that are differentially accessible to antibodies on tumor vs normal tissue, we propose that certain claudin proteins have potential as targets for novel antibody-based therapies of carcinomas.

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Abbreviations

ADCC:

Antibody-dependent cell-mediated cytotoxicity

AP:

Alkaline phosphatase

CDC:

Complement-dependent cytotoxicity

CDR:

Complementarity-determining region

CLDN:

Claudin

CPE:

Clostridium perfringens enterotoxin

dNTP:

Deoxynucleoside-triphosphate

ELISA:

Enzyme-linked immunosorbent assay

Ep-CAM:

Epithelial cell adhesion molecule

FACS:

Fluorescence-activated cell sorting

FCS:

Fetal calf serum

HPLC:

High-performance liquid chromatography

IgG:

Immunoglobulin G

KLH:

Keyhole limpet hemocyanine

NK:

Natural killer cell

PBS:

Phosphate-buffered saline

PCR:

Polymerase chain reaction

PNPP:

Paranitrophenylphosphate

RT:

Reverse transcription

TJ:

Tight junction

References

  1. Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410

    Article  CAS  PubMed  Google Scholar 

  2. Apostolopoulos V, McKenzie IF (1994) Cellular mucins: targets for immunotherapy. Crit Rev Immunol 14:293–309

    CAS  PubMed  Google Scholar 

  3. Arlt A, Vorndamm J, Muerkoster S, Yu H, Schmidt WE, Folsch UR, Schafer H (2002) Autocrine production of interleukin 1beta confers constitutive nuclear factor kappaB activity and chemoresistance in pancreatic carcinoma cell lines. Cancer Res 62:910–916

    CAS  PubMed  Google Scholar 

  4. Bhattacharjee A, Richards WG, Staunton J, Li C, Monti S, Vasa P, Ladd C, Beheshti J, Bueno R, Gillette M, Loda M, Weber G, Mark EJ, Lander ES, Wong W, Johnson BE, Golub TR, Sugarbaker DJ, Meyerson M (2001) Classification of human lung carcinomas by mRNA expression profiling reveals distinct adenocarcinoma subclasses. Proc Natl Acad Sci U S A 98:13790–13795

    Article  CAS  PubMed  Google Scholar 

  5. Braun S, Hepp F, Sommer HL, Pantel K (1999) Tumor-antigen heterogeneity of disseminated breast cancer cells: implications for immunotherapy of minimal residual disease. Int J Cancer 84:1–5

    Article  CAS  PubMed  Google Scholar 

  6. Braun S, Pantel K, Müller P, Janni W, Hepp F, Kentenich CRM, Gastroph S, Wischnik A, Dimpfl T, Kindermann G, Riethmüller G, Schlimok G (2000) Cytokeratin-positive cells in the bone marrow and survival of patients with stage I, II, or III breast cancer. N Engl J Med 342:525–533

    Article  CAS  PubMed  Google Scholar 

  7. Chester KA, Mayer A, Bhatia J, Robson L, Spencer DI, Cooke SP, Flynn AA, Sharma SK, Boxer G, Pedley RB, Begent RH (2000) Recombinant anti-carcinoembryonic antigen antibodies for targeting cancer. Cancer Chemother Pharmacol 46(Suppl): S8–S12

    CAS  PubMed  Google Scholar 

  8. Coleman M, Goldenberg DM, Siegel AB, Ketas JC, Ashe M, Fiore JM, Leonard JP (2003) Epratuzumab: targeting B-cell malignancies through CD22. Clin Cancer Res 9:3991S–3994S

    CAS  PubMed  Google Scholar 

  9. Davis DM (2002) Assembly of the immunological synapse for T cells and NK cells. Trends Immunol 23:356–363

    Article  CAS  PubMed  Google Scholar 

  10. Di Paolo C, Willuda J, Kubetzko S, Lauffer I, Tschudi D, Waibel R, Pluckthun A, Stahel RA, Zangemeister-Wittke U (2003) A recombinant immunotoxin derived from a humanized epithelial cell adhesion molecule-specific single-chain antibody fragment has potent and selective antitumor activity. Clin Cancer Res 9:2837–2848

    PubMed  Google Scholar 

  11. Fujita K, Katahira J, Horiguchi Y, Sonoda N, Furuse M, Tsukita S (2000) Clostridium perfringens enterotoxin binds to the second extracellular loop of claudin-3, a tight junction integral membrane protein. FEBS Lett 476:258–261

    Article  CAS  PubMed  Google Scholar 

  12. Furuse M, Fujita K, Hiiragi T, Fujimoto K, Tsukita S (1998) Claudin-1 and -2: novel integral membrane proteins localizing at tight junctions with no sequence similarity to occludin. J Cell Biol 141:1539–1550

    Article  CAS  PubMed  Google Scholar 

  13. Furuse M, Sasaki H, Fujimoto K, Tsukita S (1998) A single gene product, claudin-1 or -2, reconstitutes tight junction strands and recruits occludin in fibroblasts. J Cell Biol 143:391–401

    Article  CAS  PubMed  Google Scholar 

  14. Furuse M, Sasaki H, Tsukita S (1999) Manner of interaction of heterogeneous claudin species within and between tight junction strands. J Cell Biol 147:891–903

    Article  CAS  PubMed  Google Scholar 

  15. Gastl G, Spizzo G, Obrist P, Dunser M, Mikuz G (2000) Ep-CAM overexpression in breast cancer as a predictor of survival. Lancet 356:1981–1982

    Article  CAS  PubMed  Google Scholar 

  16. Ginaldi L, De Martinis M, Matutes E, Farahat N, Morilla R, Dyer MJ, Catovsky D (1998) Levels of expression of CD52 in normal and leukemic B and T cells: correlation with in vivo therapeutic responses to Campath-1H. Leuk Res 22:185–191

    Article  CAS  PubMed  Google Scholar 

  17. Goldenberg MM (1999) Trastuzumab, a recombinant DNA-derived humanized monoclonal antibody, a novel agent for the treatment of metastatic breast cancer. Clin Ther 21:309–318

    Article  CAS  PubMed  Google Scholar 

  18. Gonzalez-Mariscal L, Chavez de Ramirez B, Lazaro A, Cereijido M (1989) Establishment of tight junctions between cells from different animal species and different sealing capacities. J Membr Biol 107:43–56

    CAS  PubMed  Google Scholar 

  19. Gottlinger HG, Funke I, Johnson JP, Gokel JM, Riethmuller G (1986) The epithelial cell surface antigen 17-1A, a target for antibody-mediated tumor therapy: its biochemical nature, tissue distribution and recognition by different monoclonal antibodies. Int J Cancer 38:47–53

    CAS  PubMed  Google Scholar 

  20. Hainsworth JD (2002) Rituximab as first-line and maintenance therapy for patients with indolent non-Hodgkin’s lymphoma: interim follow-up of a multicenter phase II trial. Semin Oncol 29:25–29

    Article  CAS  Google Scholar 

  21. Hoevel T, Macek R, Mundigl O, Swisshelm K, Kubbies M (2002) Expression and targeting of the tight junction protein CLDN1 in CLDN1-negative human breast tumor cells. J Cell Physiol 191:60–68

    Article  PubMed  Google Scholar 

  22. Hough CD, Sherman-Baust CA, Pizer ES, Montz FJ, Im DD, Rosenshein NB, Cho KR, Riggins GJ, Morin PJ (2000) Large-scale serial analysis of gene expression reveals genes differentially expressed in ovarian cancer. Cancer Res 60:6281–6287

    CAS  PubMed  Google Scholar 

  23. Hughes-Jones NC, Gorick BD, Howard JC, Feinstein A (1985) Antibody density on rat red cells determines the rate of activation of the complement component C1. Eur J Immunol 15:976–980

    CAS  PubMed  Google Scholar 

  24. Katahira J, Sugiyama H, Inoue N, Horiguchi Y, Matsuda M, Sugimoto N (1997) Clostridium perfringens enterotoxin utilizes two structurally related membrane proteins as functional receptors in vivo. J Biol Chem 272:26652–26658

    Article  CAS  PubMed  Google Scholar 

  25. Khor SP, Saleh MN, Braddock JM, Pithavala YK, Thurmond L, Wissel PS, Bigley JW, LoBoglio AF (1997) A phase I trial of humanized monoclonal antibody 3622W94 including pharmacokinetics (PK) and immunogenicity evaluation. Proc Am Soc Clin Oncol. ASCO annual meeting (Abstract No. 847)

    Google Scholar 

  26. Kim JH, Herlyn D, Wong KK, Park DC, Schorge JO, Lu KH, Skates SJ, Cramer DW, Berkowitz RS, Mok SC (2003) Identification of epithelial cell adhesion molecule autoantibody in patients with ovarian cancer. Clin Cancer Res 9:4782–4791

    CAS  PubMed  Google Scholar 

  27. Kiuchi-Saishin Y, Gotoh S, Furuse M, Takasuga A, Tano Y, Tsukita S (2002) Differential expression patterns of claudins, tight junction membrane proteins, in mouse nephron segments. J Am Soc Nephrol 13:875–886

    CAS  PubMed  Google Scholar 

  28. Krogh A, Larsson B, von Heijne G, Sonnhammer EL (2001) Predicting transmembrane protein topology with a hidden Markov model: application to complete genomes. J Mol Biol 305:567–580

    Article  CAS  PubMed  Google Scholar 

  29. Kubota K, Furuse M, Sasaki H, Sonoda N, Fujita K, Nagafuchi A, Tsukita S (1999) Ca(2+)-independent cell-adhesion activity of claudins, a family of integral membrane proteins localized at tight junctions. Curr Biol 9:1035–1038

    Article  CAS  PubMed  Google Scholar 

  30. Litvinov SV, Velders MP, Bakker HA, Fleuren GJ, Warnaar SO (1994) Ep-CAM: a human epithelial antigen is a homophilic cell–cell adhesion molecule. J Cell Biol 125:437–446

    Article  CAS  PubMed  Google Scholar 

  31. Litvinov SV, Balzar M, Winter MJ, Bakker HA, Briaire-de Bruijn IH, Prins F, Fleuren GJ, Warnaar SO (1997) Epithelial cell adhesion molecule (Ep-CAM) modulates cell–cell interactions mediated by classic cadherins. J Cell Biol 139:1337–1348

    Article  CAS  PubMed  Google Scholar 

  32. LoBuglio AF, Saleh MN, Braddock JM, Lampkin TA, Khor SP, Wissel PS, Bigley JW (1997) A phase I trial of humanized anti-EGP40 monoclonal antibody 3622W94. Proc Am Soc Clin Oncol 16:436

    Google Scholar 

  33. Long H, Crean CD, Lee WH, Cummings OW, Gabig TG (2001) Expression of Clostridium perfringens enterotoxin receptors claudin-3 and claudin-4 in prostate cancer epithelium. Cancer Res 61:7878–7881

    CAS  PubMed  Google Scholar 

  34. Mack M, Riethmuller G, Kufer P (1995) A small bispecific antibody construct expressed as a functional single-chain molecule with high tumor cell cytotoxicity. Proc Natl Acad Sci U S A 92:7021–7025

    CAS  PubMed  Google Scholar 

  35. Madara JL (1998) Regulation of the movement of solutes across tight junctions. Annu Rev Physiol 60:143–159

    Article  CAS  PubMed  Google Scholar 

  36. Matter K, Balda MS (1999) Occludin and the functions of tight junctions. Int Rev Cytol 186:117–146

    CAS  PubMed  Google Scholar 

  37. Mayer A, Tsiompanou E, O’Malley D, Boxer GM, Bhatia J, Flynn AA, Chester KA, Davidson BR, Lewis AA, Winslet MC, Dhillon AP, Hilson AJ, Begent RH (2000) Radioimmunoguided surgery in colorectal cancer using a genetically engineered anti-CEA single-chain Fv antibody. Clin Cancer Res 6:1711–1719

    CAS  PubMed  Google Scholar 

  38. McCann FE, Suhling K, Carlin LM, Eleme K, Taner SB, Yanagi K, Vanherberghen B, French PM, Davis DM (2002) Imaging immune surveillance by T cells and NK cells. Immunol Rev 189:179–192

    Article  CAS  PubMed  Google Scholar 

  39. McLaughlin PM, Harmsen MC, Dokter WH, Kroesen BJ, van der Molen H, Brinker MG, Hollema H, Ruiters MH, Buys CH, de Leij LF (2001) The epithelial glycoprotein 2 (EGP-2) promoter-driven epithelial-specific expression of EGP-2 in transgenic mice: a new model to study carcinoma-directed immunotherapy. Cancer Res 61:4105–4111

    CAS  PubMed  Google Scholar 

  40. Michl P, Buchholz M, Rolke M, Kunsch S, Lohr M, McClane B, Tsukita S, Leder G, Adler G, Gress TM (2001) Claudin-4: a new target for pancreatic cancer treatment using Clostridium perfringens enterotoxin. Gastroenterology 121:678–684

    CAS  PubMed  Google Scholar 

  41. Mitic LL, Van Itallie CM, Anderson JM (2000) Molecular physiology and pathophysiology of tight junctions I. Tight junction structure and function: lessons from mutant animals and proteins. Am J Physiol Gastrointest Liver Physiol 279:G250–G254

    CAS  PubMed  Google Scholar 

  42. Morita K, Furuse M, Fujimoto K, Tsukita S (1999) Claudin multigene family encoding four-transmembrane domain protein components of tight junction strands. Proc Natl Acad Sci U S A 96:511–516

    Article  CAS  PubMed  Google Scholar 

  43. Morita K, Sasaki H, Furuse M, Tsukita S (1999) Endothelial claudin: claudin-5/TMVCF constitutes tight junction strands in endothelial cells. J Cell Biol 147:185–194

    Article  CAS  PubMed  Google Scholar 

  44. Naundorf S, Preithner S, Mayer P, Lippold S, Wolf A, Hanakam F, Fichtner I, Kufer P, Raum T, Riethmuller G, Baeuerle PA, Dreier T (2002) In vitro and in vivo activity of MT201, a fully human monoclonal antibody for pancarcinoma treatment. Int J Cancer 100:101–110

    Article  CAS  PubMed  Google Scholar 

  45. Ogura E, Senzaki H, Yoshizawa K, Hioki K, Tsubura A (1998) Immunohistochemical localization of epithelial glycoprotein EGP-2 and carcinoembryonic antigen in normal colonic mucosa and colorectal tumors. Anticancer Res 18:3669–3675

    CAS  PubMed  Google Scholar 

  46. Pantel K, Izbicki J, Passlick B, Angstwurm M, Haussinger K, Thetter O, Riethmuller G (1996) Frequency and prognostic significance of isolated tumour cells in bone marrow of patients with non-small-cell lung cancer without overt metastases. Lancet 347:649–653

    Article  CAS  PubMed  Google Scholar 

  47. Piyathilake CJ, Frost AR, Weiss H, Manne U, Heimburger DC, Grizzle WE (2000) The expression of Ep-CAM (17-1A) in squamous cell cancers of the lung. Hum Pathol 31:482–487

    Article  CAS  PubMed  Google Scholar 

  48. Poczatek RB, Myers RB, Manne U, Oelschlager DK, Weiss HL, Bostwick DG, Grizzle WE (1999) Ep-Cam levels in prostatic adenocarcinoma and prostatic intraepithelial neoplasia. J Urol 162:1462–1466

    Article  CAS  PubMed  Google Scholar 

  49. Rahner C, Mitic LL, Anderson JM (2001) Heterogeneity in expression and subcellular localization of claudins 2, 3, 4, and 5 in the rat liver, pancreas, and gut. Gastroenterology 120:411–422

    CAS  PubMed  Google Scholar 

  50. Riethmuller G, Klein CA, Pantel K (1999) Hunting down the seminal cells of clinical metastases. Immunol Today 20:294–296

    Article  CAS  PubMed  Google Scholar 

  51. Rossmann ED, Lundin J, Lenkei R, Mellstedt H, Osterborg A (2001) Variability in B-cell antigen expression: implications for the treatment of B-cell lymphomas and leukemias with monoclonal antibodies. Hematol J 2:300–306

    Article  CAS  PubMed  Google Scholar 

  52. Saleh MN, Khazaeli LM, Thurmond LM, Khor SP, Lampkin TA, Wissel PS, LoBouglio AF (1998) Phase I trial testing multiple doses of humanized monoclonal antibody (Mab) 3622W94. Proc Am Soc Clin Oncol. ASCO annual meeting (Abstract No. 1680)

    Google Scholar 

  53. Schwartzberg LS (2001) Clinical experience with edrecolomab: a monoclonal antibody therapy for colorectal carcinoma. Crit Rev Oncol Hematol 40:17–24

    Article  CAS  PubMed  Google Scholar 

  54. Sondel PM, Hank JA, Gan J, Neal Z, Albertini MR (2003) Preclinical and clinical development of immunocytokines. Curr Opin Investig Drugs 4:696–700

    CAS  PubMed  Google Scholar 

  55. Sonoda N, Furuse M, Sasaki H, Yonemura S, Katahira J, Horiguchi Y, Tsukita S (1999) Clostridium perfringens enterotoxin fragment removes specific claudins from tight junction strands: evidence for direct involvement of claudins in tight junction barrier. J Cell Biol 147:195–204

    Article  PubMed  Google Scholar 

  56. Spizzo G, Obrist P, Ensinger C, Theurl I, Dunser M, Ramoni A, Gunsilius E, Eibl G, Mikuz G, Gastl G (2002) Prognostic significance of Ep-CAM AND Her-2/neu overexpression in invasive breast cancer. Int J Cancer 98:883–888

    Article  CAS  PubMed  Google Scholar 

  57. Steffens MG, Boerman OC, Oosterwijk-Wakka JC, Oosterhof GO, Witjes JA, Koenders EB, Oyen WJ, Buijs WC, Debruyne FM, Corstens FH, Oosterwijk E (1997) Targeting of renal cell carcinoma with iodine-131-labeled chimeric monoclonal antibody G250. J Clin Oncol 15:1529–1537

    CAS  PubMed  Google Scholar 

  58. Terris B, Blaveri E, Crnogorac-Jurcevic T, Jones M, Missiaglia E, Ruszniewski P, Sauvanet A, Lemoine NR (2002) Characterization of gene expression profiles in intraductal papillary-mucinous tumors of the pancreas. Am J Pathol 160:1745–1754

    CAS  PubMed  Google Scholar 

  59. Tsukita S, Furuse M (2000) Pores in the wall: claudins constitute tight junction strands containing aqueous pores. J Cell Biol 149:13–16

    Article  CAS  PubMed  Google Scholar 

  60. Tsukita S, Furuse M (2002) Claudin-based barrier in simple and stratified cellular sheets. Curr Opin Cell Biol 14:531–536

    Article  CAS  PubMed  Google Scholar 

  61. Tsukita S, Furuse M, Itoh M (1999) Structural and signalling molecules come together at tight junctions. Curr Opin Cell Biol 11:628–633

    Article  CAS  PubMed  Google Scholar 

  62. Velders MP, Litvinov SV, Warnaar SO, Gorter A, Fleuren GJ, Zurawski VR Jr, Coney LR (1994) New chimeric anti-pancarcinoma monoclonal antibody with superior cytotoxicity-mediating potency. Cancer Res 54:1753–1759

    CAS  PubMed  Google Scholar 

  63. Weckermann D, Muller P, Wawroschek F, Harzmann R, Riethmuller G, Schlimok G (2001) Disseminated cytokeratin positive tumor cells in the bone marrow of patients with prostate cancer: detection and prognostic value. J Urol 166:699–703

    Article  CAS  PubMed  Google Scholar 

  64. Wilcox ER, Burton QL, Naz S, Riazuddin S, Smith TN, Ploplis B, Belyantseva I, Ben-Yosef T, Liburd NA, Morell RJ, Kachar B, Wu DK, Griffith AJ, Friedman TB (2001) Mutations in the gene encoding tight junction claudin-14 cause autosomal recessive deafness DFNB29. Cell 104:165–172

    Article  CAS  PubMed  Google Scholar 

  65. Zhang S, Zhang HS, Reuter VE, Slovin SF, Scher HI, Livingston PO (1998) Expression of potential target antigens for immunotherapy on primary and metastatic prostate cancers. Clin Cancer Res 4:295–302

    CAS  PubMed  Google Scholar 

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Acknowledgements

We are grateful to Maria Lahme for technical assistance and to Silvia Krieg for assistance in preparing the manuscript.

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Authors and Affiliations

  1. Micromet AG, Staffelseestr. 2, 81477, Munich, Germany

    Sonja Offner, Ulrike Teichmann, Susanne Weinberger, Susanne Gross, Peter Kufer, Christian Itin, Patrick A. Baeuerle & Birgit Kohleisen

  2. Department of Microbiology and Immunology, University of California, Box 0414, San Francisco, California, USA

    Armin Hekele

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  1. Sonja Offner
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Offner, S., Hekele, A., Teichmann, U. et al. Epithelial tight junction proteins as potential antibody targets for pancarcinoma therapy. Cancer Immunol Immunother 54, 431–445 (2005). https://doi.org/10.1007/s00262-004-0613-x

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  • DOI: https://doi.org/10.1007/s00262-004-0613-x

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