Cancer Immunology, Immunotherapy

, Volume 58, Issue 8, pp 1185–1194

Breast cancer cells expressing stem cell markers CD44+ CD24lo are eliminated by Numb-1 peptide-activated T cells

  • Takashi Mine
  • Satoko Matsueda
  • Yufeng Li
  • Hiroshi Tokumitsu
  • Hui Gao
  • Cristopher Danes
  • Kwong-Kwok Wong
  • Xinhui Wang
  • Soldano Ferrone
  • Constantin G. Ioannides
Original Article

Abstract

Cancer stem cells (CSC) are resistant to chemo- and radiotherapy. To eliminate cells with phenotypic markers of CSC-like we characterized: (1) expression of CD44, CD24, CD133 and MIC-A/B (NKG2 receptors) in breast (MCF7) and ovarian (SK-OV-3) cells resistant to gemcitabine (GEM), paclitaxel (PTX) and 5-fluorouracil (5-FU) and (2) their elimination by Numb- and Notch-peptide activated CTL. The number of cells in all populations with the luminal CSC phenotype [epithelial specific antigen+ (ESA) CD44hi CD24lo, CD44hi CD133+, and CD133+ CD24lo] increased in drug-resistant MCF7 and SK-OV-3 cells. Similarly, the number of cells with expressed MIC-A/B increased 4 times in drug-resistant tumor cells compared with drug-sensitive cells. GEMRes MCF7 cells had lower levels of the Notch-1-extracellular domain (NECD) and Notch trans-membrane intracellular domain (TMIC) than GEMSens MCF7. The levels of Numb, and Numb-L-[P]-Ser265 were similar in GEMRes and GEMSens MCF7 cells. Only the levels of Numb-L (long)-Ser295 decreased slightly. This finding suggests that Notch-1 cleavage to TMIC is inhibited in GEMRes MCF7 cells. PBMC activated by natural immunogenic peptides Notch-1 (2112–2120) and Numb-1 (87–95) eliminated NICDpositive, CD24hi CD24lo MCF7 cells. It is likely that the immunogenic Numb-1 peptide in MCF7 cells originated from Numb, [P]-lated by an unknown kinase, because staurosporine but not wortmannin and MAPK-inhibitors decreased peptide presentation. Numb and Notch are antagonistic proteins which degrade each other to stop and activate cell proliferation, respectively. Their peptides are presented alternatively. Targeting both antagonistic proteins should be useful to prevent metastases in patients whose tumors are resistant to conventional treatments.

Keywords

Drug resistance Breast/ovarian Cancer stem cell Notch Numb Peptide 

Supplementary material

262_2008_623_MOESM1_ESM.pdf (54 kb)
Supplementary material (PDF 54 kb)

References

  1. 1.
    Al-Hajj M, Wicha MS, Benito-Hernandez A, Morrison SJ, Clarke MF (2003) Prospective identification of tumorigenic breast cancer cells. Proc Natl Acad Sci USA 100:3983–3988PubMedCrossRefGoogle Scholar
  2. 2.
    Androutsellis-Theotokis A, Leker RR, Soldner F, Hoeppner DJ, Ravin R, Poser SW, Rueger MA, Bae SK, Kittappa R, McKay RD (2006) Notch signaling regulates stem cell numbers in vitro and in vivo. Nature 442:823–826PubMedCrossRefGoogle Scholar
  3. 3.
    Balic M, Lin H, Young L, Hawes D, Giuliano A, McNamara G, Datar RH, Cote RJ (2006) Most early disseminated cancer cells detected in bone marrow of breast cancer patients have a putative breast cancer stem cell phenotype. Clin Cancer Res 12:5615–5621PubMedCrossRefGoogle Scholar
  4. 4.
    Berdnik D, Torok T, Gonzalez-Gaitan M, Knoblich JA (2002) The endocytic protein a-adaptin is required for numb-mediated asymmetric cell division in Drosophila. Dev Cell 3:221–231PubMedCrossRefGoogle Scholar
  5. 5.
    Blaumueller CM, Qi H, Zagouras P, Artavanis-Tsakonas S (1997) Intracellular cleavage of Notch leads to a heterodimeric receptor on the plasma membrane. Cell 90(2):281–291PubMedCrossRefGoogle Scholar
  6. 6.
    Bourguignon LY, Gilad E, Peyrollier K (2007) Heregulin-mediated ErbB2-ERK signaling activates hyaluronan synthases leading to CD44-dependent ovarian tumor cell growth and migration. J Biol Chem 282:19426–19441PubMedCrossRefGoogle Scholar
  7. 7.
    Cariati M, Naderi A, Brown JP, Smalley MJ, Pinder SE, Caldas C, Purushotham AD (2008) ã6 integrin is necessary for the tumourigenicity of a stem cell-like subpopulation within the MCF7 breast cancer cell line. Int J Cancer 122(2):298–304PubMedCrossRefGoogle Scholar
  8. 8.
    Castilleja A, Carter D, Efferson CL, Ward NE, Kawano K, Fisk B, Kudelka AP, Gershenson DM, Murray JL, O’Brian CA, Ioannides CG (2002) Induction of tumor-reactive CTL by C-side chain variants of the CTL epitope HER-2/neu proto-oncogene (369–377) selected by molecular modeling of the peptide: HLA-A2 complex. J Immunol 169:3545–3554PubMedGoogle Scholar
  9. 9.
    Chapman G, Liu L, Sahlgren C, Dahlqvist C, Lendahl U (2006) High levels of Notch signaling down-regulate Numb and Numblike. J Cell Biol 175:535–540PubMedCrossRefGoogle Scholar
  10. 10.
    Collins AT, Berry PA, Hyde C, Stower MJ, Maitland NJ (2005) Prospective identification of tumorigenic prostate cancer stem cells. Cancer Res 65:10946–10951PubMedCrossRefGoogle Scholar
  11. 11.
    Di Marcotullio L, Ferretti E, Greco A, De Smaele E, Po A, Sico MA, Alimandi M, Giannini G, Maroder M, Screpanti I, Gulino A (2006) Numb is a suppressor of Hedgehog signalling and targets Gli1 for Itch-dependent ubiquitination. Nat Cell Biol 8:1415–1423PubMedCrossRefGoogle Scholar
  12. 12.
    Edwards SL, Brough R, Lord CJ, Natrajan R, Vatcheva R, Levine DA, Boyd J, Reis-Filho JS, Ashworth (2008) A resistance to therapy caused by intragenic deletion in BRCA2. Nature 451:1111–1115PubMedCrossRefGoogle Scholar
  13. 13.
    Efferson CL, Tsuda N, Kawano K, Nistal-Villan E, Sellappan S, Yu D, Murray JL, Garcia-Sastre A, Ioannides CG (2006) Prostate tumor cells infected with a recombinant influenza virus expressing a truncated NS1 protein activate cytolytic CD8 + cells to recognize noninfected tumor cells. J Virol 80:383–394PubMedCrossRefGoogle Scholar
  14. 14.
    Hurlbut GD, Kankel MW, Lake RJ, Artavanis-Tsakonas S (2007) Crossing paths with Notch in the hyper-network. Curr Opin Cell Biol 2:166–175CrossRefGoogle Scholar
  15. 15.
    Ishiyama S, Matsueda S, Jones LA, Efferson C, Celestino J, Schmandt R, Ioannides CG, Tsuda N, Chang DZ (2007) Novel natural immunogenic peptides from Numb1 and Notch1 proteins for CD8+ cells in ovarian ascites. Int J Oncol 30:889–898PubMedGoogle Scholar
  16. 16.
    Kawano K, Efferson CL, Peoples GE, Carter D, Tsuda N, Murray JL, Ioannides CG (2005) Sensitivity of undifferentiated, high-TCR density CD8 + cells to methylene groups appended to tumor antigen determines their differentiation or death. Cancer Res 65:2930–2937PubMedCrossRefGoogle Scholar
  17. 17.
    Li X, Lewis MT, Huang J, Gutierrez C, Osborne CK, Wu MF, Hilsenbeck SG, Pavlick A, Zhang X, Chamness GC, Wong H, Rosen J, Chang JC (2008) Intrinsic resistance of tumorigenic breast cancer cells to chemotherapy. J Natl Cancer Inst 100(9):672–679PubMedCrossRefGoogle Scholar
  18. 18.
    Liu S, Ginestier C, Charafe-Jauffret E, Foco H, Kleer CG, Merajver SD, Dontu G, Wicha MS (2008) BRCA1 regulates human mammary stem/progenitor cell fate. Proc Natl Acad Sci USA 105(5):1680–1685PubMedCrossRefGoogle Scholar
  19. 19.
    McGill MA, McGlade CJ (2003) Mammalian Numb proteins promote Notch1 receptor ubiquitination and degradation of the Notch1 intracellular domain. J Biol Chem 278:23196–23203PubMedCrossRefGoogle Scholar
  20. 20.
    Morrison SJ, Kimble J (2006) Asymmetric and symmetric stem-cell divisions in development and cancer. Nature 441:1068–1074PubMedCrossRefGoogle Scholar
  21. 21.
    Mumm JS, Schroeter EH, Saxena MT, Griesemer A, Tian X, Pan DJ, Ray WJ, Kopan RA (2000) Ligand-induced extracellular cleavage regulates gamma-secretase-like proteolytic activation of Notch1. Mol Cell 5(2):197–206PubMedCrossRefGoogle Scholar
  22. 22.
    Nishimura T, Kaibuchi K (2007) Numb controls integrin endocytosis for directional cell migration with aPKC and PAR-3. Dev Cell 1:15–28CrossRefGoogle Scholar
  23. 23.
    Orford KW, Scadden DT (2008) Deconstructing stem cell self-renewal: genetic insights into cell-cycle regulation. Nat Rev Genet 9:115–128PubMedCrossRefGoogle Scholar
  24. 24.
    Ouhtit A, Abd Elmageed ZY, Abdraboh ME, Lioe TF, Raj MH (2007) In vivo evidence for the role of CD44 in promoting breast cancer metastasis to the liver. Am J Pathol 171(6):2033–2039PubMedCrossRefGoogle Scholar
  25. 25.
    Phillips TM, McBride WH, Pajonk F (2006) The response of CD24(-/low)/CD44 + breast cancer-initiating cells to radiation. J Natl Cancer Inst 98(24):1777–1785PubMedCrossRefGoogle Scholar
  26. 26.
    Piekarz RL, Sackett DL, Bates SE (2007) Histone deacetylase inhibitors and demethylating agents: clinical development of histone deacetylase inhibitors for cancer therapy. Cancer J 13(1):30–39PubMedCrossRefGoogle Scholar
  27. 27.
    Ratnam K, Low JA (2007) Current development of clinical inhibitors of poly (ADP-ribose)-polymerase in oncology. Clin Cancer Res 13:1383–1388PubMedCrossRefGoogle Scholar
  28. 28.
    Ricci-Vitiani L, Lombardi DG, Pilozzi E, Biffoni M, Todaro M, Peschle C, De Maria R (2007) Identification and expansion of human colon-cancer-initiating cells. Nature 445:111–115PubMedCrossRefGoogle Scholar
  29. 29.
    Sansone P, Storci G, Tavolari S, Guarnieri T, Giovannini C, Taffurelli M, Ceccarelli C, Santini D, Paterini P, Marcu KB, Chieco P, Bonafè M (2007) IL-6 triggers malignant features in mammospheres from human ductal breast carcinoma and normal mammary gland. J Clin Invest 117:3988–4002PubMedCrossRefGoogle Scholar
  30. 30.
    Schroeter EH, Kisslinger JA, Kopan R (1998) Notch-1 signalling requires ligand-induced proteolytic release of intracellular domain. Nature 393(6683):382–386PubMedCrossRefGoogle Scholar
  31. 31.
    Singh SK, Clarke ID, Terasaki M, Bonn VE, Hawkins C, Squire J, Dirks PB (2003) Identification of a cancer stem cell in human brain tumors. Cancer Res 63:5821–5828PubMedGoogle Scholar
  32. 32.
    Smith CA, Lau KM, Rahmani Z, Dho SE, Brothers G, She YM, Berry DM, Bonneil E, Thibault P, Schweisguth F, Le Borgne R, McGlade CJ (2007) aPKC-mediated phosphorylation regulates asymmetric membrane localization of the cell fate determinant Numb. EMBO J 26:468–480PubMedCrossRefGoogle Scholar
  33. 33.
    Surowiak P, Materna V, Kaplenko I, Spaczyński M, Dietel M, Kristiansen G, Lage H, Zabel M (2006) Unfavorable prognostic value of CD24 expression in sections from primary and relapsed ovarian cancer tissue. Int J Gynecol Cancer 2:515–521CrossRefGoogle Scholar
  34. 34.
    Tokumitsu H, Hatano N, Inuzuka H, Sueyoshi Y, Yokokura S, Ichimura T, Nozaki N, Kobayashi R (2005) Phosphorylation of Numb family proteins. Possible involvement of Ca2 +/calmodulin-dependent protein kinases. J Biol Chem 280:35108–35118PubMedCrossRefGoogle Scholar
  35. 35.
    Tokumitsu H, Hatano N, Yokokura S, Sueyoshi Y, Nozaki N, Kobayashi R (2006) Phosphorylation of Numb regulates its interaction with the clathrin-associated adaptor AP-2. FEBS Lett 580(24):5797–5801PubMedCrossRefGoogle Scholar
  36. 36.
    Tolis C, Peters GJ, Ferreira CG, Pinedo HM, Giaccone G (1999) Cell cycle disturbances and apoptosis induced by topotecan and gemcitabine on human lung cancer cell lines. Eur J Cancer 35:796–807PubMedCrossRefGoogle Scholar
  37. 37.
    Tsuda N, Chang DZ, Mine T, Efferson C, García-Sastre A, Wang X, Ferrone S, Ioannides CG (2007) Taxol increases the amount and T cell activating ability of self-immune stimulatory multimolecular complexes found in ovarian cancer cells. Cancer Res 67:8378–8387PubMedCrossRefGoogle Scholar
  38. 38.
    Wang H, Ouyang Y, Somers WG, Chia W, Lu B (2007) Polo inhibits progenitor self-renewal and regulates Numb asymmetry by phosphorylating Pon. Nature 449(7158):96–100PubMedCrossRefGoogle Scholar
  39. 39.

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Takashi Mine
    • 1
    • 2
    • 9
  • Satoko Matsueda
    • 1
  • Yufeng Li
    • 1
    • 6
  • Hiroshi Tokumitsu
    • 3
  • Hui Gao
    • 4
  • Cristopher Danes
    • 5
  • Kwong-Kwok Wong
    • 1
  • Xinhui Wang
    • 7
  • Soldano Ferrone
    • 7
  • Constantin G. Ioannides
    • 1
    • 8
  1. 1.Department of Gynecologic OncologyThe University of Texas M. D. Anderson Cancer CenterHoustonUSA
  2. 2.Departments of Immunology and SurgeryKurume University School of MedicineKurumeJapan
  3. 3.Department of Cell Signaling, Faculty of MedicineKagawa UniversityKagawaJapan
  4. 4.Department of Molecular PathologyThe University of Texas M. D. Anderson Cancer CenterHoustonUSA
  5. 5.Department of Molecular and Cellular OncologyThe University of Texas M. D. Anderson Cancer CenterHoustonUSA
  6. 6.Department of Melanoma Medical OncologyThe University of Texas M. D. Anderson Cancer CenterHoustonUSA
  7. 7.Departments of Surgery, Immunology and PathologyUniversity of Pittsburgh Cancer InstitutePittsburghUSA
  8. 8.Department of ImmunologyThe University of Texas M. D. Anderson Cancer CenterHoustonUSA
  9. 9.Multidisciplinary Treatment CenterKurume University School of MedicineKurumeJapan

Personalised recommendations