The MUC1 tumor associated antigen is highly expressed on a range of tumors. Its broad distribution on primary tumors and metastases renders it an attractive target for immunotherapy. After synthesis MUC1 is cleaved, yielding a large soluble extracellular alpha subunit containing the tandem repeats array (TRA) domain specifically bound, via non-covalent interaction, to a smaller beta subunit containing the transmembrane and cytoplasmic domains. Thus far, inconclusive efficacy has been reported for anti-MUC1 antibodies directed against the soluble alpha subunit. Targeting the cell bound beta subunit, may bypass limitations posed by circulating TRA domains. MUC1’s signal peptide (SP) domain promiscuously binds multiple MHC class II and Class I alleles, which upon vaccination, generated robust T-cell immunity against MUC1-positive tumors. This is a first demonstration of non-MHC associated, MUC1 specific, cell surfaces presence for MUC1 SP domain. Polyclonal and monoclonal antibodies generated against MUC1 SP domain specifically bind a large variety of MUC1-positive human solid and haematological tumor cell lines; MUC1-positive bone marrow derived plasma cells obtained from multiple myeloma (MM)-patients, but not MUC1 negative tumors cells, and normal naive primary blood and epithelial cells. Membranal MUC1 SP appears mainly as an independent entity but also co-localized with the full MUC1 molecule. MUC1-SP specific binding in BM-derived plasma cells can assist in selecting patients to be treated with anti-MUC1 SP therapeutic vaccine, ImMucin. A therapeutic potential of the anti-MUC1 SP antibodies was suggested by their ability to support of complement-mediated lysis of MUC1-positive tumor cells but not MUC1 negative tumor cells and normal naive primary epithelial cells. These findings suggest a novel cell surface presence of MUC1 SP domain, a potential therapeutic benefit for anti-MUC1 SP antibodies in MUC1-positive tumors and a selection tool for MM patients to be treated with the anti-MUC1 SP vaccine, ImMucin.
References
[1]
Apostolopoulos V, McKenzie IF (1994) Cellular mucins: targets for immunotherapy. Crit Rev Immunol 14: 293–309.
[2]
Brossart P, Schneider A, Dill P, Schammann T, Grunebach F, et al. (2001) The epithelial tumor antigen MUC1 is expressed in hematological malignancies and is recognized by MUC1-specific cytotoxic T-lymphocytes. Cancer Res 61: 6846–6850.
[3]
Lacunza E, Baudis M, Colussi AG, Segal-Eiras A, Croce MV, et al. (2010) MUC1 oncogene amplification correlates with protein overexpression in invasive breast carcinoma cells. Cancer Genet Cytogenet 201: 102–110.
[4]
McGuckin MA, Walsh MD, Hohn BG, Ward BG, Wright RG (1995) Prognostic significance of MUC1 epithelial mucin expression in breast cancer. Hum Pathol 26: 432–439.
[5]
Ho SB, Niehans GA, Lyftogt C, Yan PS, Cherwitz DL, et al. (1993) Heterogeneity of mucin gene expression in normal and neoplastic tissues. Cancer Res 53: 641–651.
[6]
Segev N, Hizi A, Kirenberg F, Keydar I (1985) Characterization of a protein, released by the T47D cell line, immunologically related to the major envelope protein of mouse mammary tumor virus. Proceedings of the National Academy of Sciences of the United States of America 82: 1531–1535.
[7]
Engelmann K, Shen H, Finn OJ (2008) MCF7 side population cells with characteristics of cancer stem/progenitor cells express the tumor antigen MUC1. Cancer Res 68: 2419–2426.
[8]
Graham RA, Burchell JM, Taylor-Papadimitriou J (1996) The polymorphic epithelial mucin: potential as an immunogen for a cancer vaccine. Cancer Immunol Immunother 42: 71–80.
[9]
Tang CK, Katsara M, Apostolopoulos V (2008) Strategies used for MUC1 immunotherapy: human clinical studies. Expert Rev Vaccines 7: 963–975.
[10]
Treon SP, Raje N, Anderson KC (2000) Immunotherapeutic strategies for the treatment of plasma cell malignancies. Semin Oncol 27: 598–613.
[11]
Cheever MA, Allison JP, Ferris AS, Finn OJ, Hastings BM, et al. (2009) The prioritization of cancer antigens: a national cancer institute pilot project for the acceleration of translational research. Clinical cancer research : an official journal of the American Association for Cancer Research 15: 5323–5337.
[12]
Baruch A, Hartmann M, Zrihan-Licht S, Greenstein S, Burstein M, et al. (1997) Preferential expression of novel MUC1 tumor antigen isoforms in human epithelial tumors and their tumor-potentiating function. International journal of cancer Journal international du cancer 71: 741–749.
[13]
Hareuveni M, Tsarfaty I, Zaretsky J, Kotkes P, Horev J, et al. (1990) A transcribed gene, containing a variable number of tandem repeats, codes for a human epithelial tumor antigen. cDNA cloning, expression of the transfected gene and over-expression in breast cancer tissue. Eur J Biochem 189: 475–486.
[14]
Gendler SJ, Lancaster CA, Taylor-Papadimitriou J, Duhig T, Peat N, et al. (1990) Molecular cloning and expression of human tumor-associated polymorphic epithelial mucin. The Journal of biological chemistry 265: 15286–15293.
[15]
Ligtenberg MJ, Kruijshaar L, Buijs F, van Meijer M, Litvinov SV, et al. (1992) Cell-associated episialin is a complex containing two proteins derived from a common precursor. The Journal of biological chemistry 267: 6171–6177.
[16]
Danielczyk A, Stahn R, Faulstich D, Loffler A, Marten A, et al. (2006) PankoMab: a potent new generation anti-tumour MUC1 antibody. Cancer Immunol Immunother 55: 1337–1347.
[17]
Thie H, Toleikis L, Li J, von Wasielewski R, Bastert G, et al. (2011) Rise and fall of an anti-MUC1 specific antibody. PLoS One 6: e15921.
[18]
Fan XN, Karsten U, Goletz S, Cao Y (2010) Reactivity of a humanized antibody (hPankoMab) towards a tumor-related MUC1 epitope (TA-MUC1) with various human carcinomas. Pathol Res Pract 206: 585–589.
[19]
Agrawal B, Krantz MJ, Reddish MA, Longenecker BM (1998) Cancer-associated MUC1 mucin inhibits human T-cell proliferation, which is reversible by IL-2. Nat Med 4: 43–49.
[20]
Brossart P, Heinrich KS, Stuhler G, Behnke L, Reichardt VL, et al. (1999) Identification of HLA-A2-restricted T-cell epitopes derived from the MUC1 tumor antigen for broadly applicable vaccine therapies. Blood 93: 4309–4317.
[21]
Carmon L, El-Shami KM, Paz A, Pascolo S, Tzehoval E, et al. (2000) Novel breast-tumor-associated MUC1-derived peptides: characterization in Db？/？ x beta2 microglobulin (beta2m) null mice transgenic for a chimeric HLA-A2.1/Db-beta2 microglobulin single chain. Int J Cancer 85: 391–397.
[22]
Fung PY, Longenecker BM (1991) Specific immunosuppressive activity of epiglycanin, a mucin-like glycoprotein secreted by a murine mammary adenocarcinoma (TA3-HA). Cancer Res 51: 1170–1176.
[23]
Gillespie AM, Broadhead TJ, Chan SY, Owen J, Farnsworth AP, et al. (2000) Phase I open study of the effects of ascending doses of the cytotoxic immunoconjugate CMB-401 (hCTMO1-calicheamicin) in patients with epithelial ovarian cancer. Annals of oncology : official journal of the European Society for Medical Oncology/ESMO 11: 735–741.
[24]
Pichinuk E, Benhar I, Jacobi O, Chalik M, Weiss L, et al. (2012) Antibody targeting of cell-bound MUC1 SEA domain kills tumor cells. Cancer Res 72: 3324–3336.
[25]
Prinssen HM, Molthoff CF, Verheijen RH, Broadhead TJ, Kenemans P, et al. (1998) Biodistribution of 111In-labelled engineered human antibody CTM01 (hCTM01) in ovarian cancer patients: influence of prior administration of unlabelled hCTM01. Cancer immunology, immunotherapy: CII 47: 39–46.
[26]
van de Wiel-van Kemenade E, Ligtenberg MJ, de Boer AJ, Buijs F, Vos HL, et al. (1993) Episialin (MUC1) inhibits cytotoxic lymphocyte-target cell interaction. J Immunol 151: 767–776.
[27]
Correa I, Plunkett T, Coleman J, Galani E, Windmill E, et al. (2005) Responses of human T cells to peptides flanking the tandem repeat and overlapping the signal sequence of MUC1. International journal of cancer Journal international du cancer 115: 760–768.
[28]
Kovjazin R, Volovitz I, Kundel Y, Rosenbaum E, Medalia G, et al. (2011) ImMucin: a novel therapeutic vaccine with promiscuous MHC binding for the treatment of MUC1-expressing tumors. Vaccine 29: 4676–4686.
[29]
Martoglio B, Dobberstein B (1998) Signal sequences: more than just greasy peptides. Trends Cell Biol 8: 410–415.
[30]
Henderson RA, Michel H, Sakaguchi K, Shabanowitz J, Appella E, et al. (1992) HLA-A2.1-associated peptides from a mutant cell line: a second pathway of antigen presentation. Science 255: 1264–1266.
[31]
Kovjazin R, Volovitz I, Daon Y, Vider-Shalit T, Azran R, et al. (2011) Signal peptides and trans-membrane regions are broadly immunogenic and have high CD8+ T cell epitope densities: Implications for vaccine development. Mol Immunol 48: 1009–1018.
[32]
Lyko F, Martoglio B, Jungnickel B, Rapoport TA, Dobberstein B (1995) Signal sequence processing in rough microsomes. J Biol Chem 270: 19873–19878.
[33]
Martoglio B (2003) Intramembrane proteolysis and post-targeting functions of signal peptides. Biochem Soc Trans 31: 1243–1247.
[34]
Kovjazin R, Dubnik A, Horn G, Smorodinsky NI, Hardan I, et al. (2012) Autoantibodies against the signal peptide domain of MUC1 in patients with multiple myeloma: Implications for disease diagnosis and prognosis. Exp Ther Med 3: 1092–1098.
[35]
Keydar I, Chou CS, Hareuveni M, Tsarfaty I, Sahar E, et al. (1989) Production and characterization of monoclonal antibodies identifying breast tumor-associated antigens. Proc Natl Acad Sci U S A 86: 1362–1366.
[36]
Alausa OK (1975) The use of ammonium sulphate precipitation method for the determination of antigen-binding capacity and affinity of anti-tetanus antibodies in human serum. Journal of immunological methods 8: 117–126.
[37]
Rubinstein DB, Karmely M, Ziv R, Benhar I, Leitner O, et al. (2006) MUC1/X protein immunization enhances cDNA immunization in generating anti-MUC1 alpha/beta junction antibodies that target malignant cells. Cancer research 66: 11247–11253.
[38]
Ye RD, Wun TC, Sadler JE (1988) Mammalian protein secretion without signal peptide removal. Biosynthesis of plasminogen activator inhibitor-2 in U-937 cells. The Journal of biological chemistry 263: 4869–4875.
[39]
Palmiter RD, Gagnon J, Walsh KA (1978) Ovalbumin: a secreted protein without a transient hydrophobic leader sequence. Proceedings of the National Academy of Sciences of the United States of America 75: 94–98.
[40]
Schrempf S, Froeschke M, Giroglou T, von Laer D, Dobberstein B (2007) Signal peptide requirements for lymphocytic choriomeningitis virus glycoprotein C maturation and virus infectivity. J Virol 81: 12515–12524.
[41]
Strome SE, Sausville EA, Mann D (2007) A mechanistic perspective of monoclonal antibodies in cancer therapy beyond target-related effects. The oncologist 12: 1084–1095.
[42]
Cheung NK, Walter EI, Smith-Mensah WH, Ratnoff WD, Tykocinski ML, et al. (1988) Decay-accelerating factor protects human tumor cells from complement-mediated cytotoxicity in vitro. The Journal of clinical investigation 81: 1122–1128.
[43]
Flanders A, Stetler-Stevenson M, Landgren O (2013) Minimal residual disease testing in multiple myeloma by flow cytometry: major heterogeneity. Blood 122: 1088–1089.
[44]
Wijdenes J, Vooijs WC, Clement C, Post J, Morard F, et al. (1996) A plasmocyte selective monoclonal antibody (B-B4) recognizes syndecan-1. British journal of haematology 94: 318–323.
[45]
Nakayama S, Yokote T, Hirata Y, Iwaki K, Akioka T, et al. (2012) An approach for diagnosing plasma cell myeloma by three-color flow cytometry based on kappa/lambda ratios of CD38-gated CD138(+) cells. Diagn Pathol 7: 131.
[46]
Matsui W, Huff CA, Wang Q, Malehorn MT, Barber J, et al. (2004) Characterization of clonogenic multiple myeloma cells. Blood 103: 2332–2336.
