Abstract

Cancer-specific cell-surface antigens are ideal targets for monoclonal antibody (mAb)-based immunotherapy but are likely to have previously been identified in transcriptome or proteome analyses. Here, we show that the active conformer of an integrin can serve as a specific therapeutic target for multiple myeloma (MM). We screened >10,000 anti-MM mAb clones and identified MMG49 as an MM-specific mAb specifically recognizing a subset of integrin β7 molecules. The MMG49 epitope, in the N-terminal region of the β7 chain, is predicted to be inaccessible in the resting integrin conformer but exposed in the active conformation. Elevated expression and constitutive activation of integrin β7 conferred high MMG49 reactivity on MM cells, whereas MMG49 binding was scarcely detectable in other cell types including normal integrin β7+ lymphocytes. T cells transduced with MMG49-derived chimeric antigen receptor (CAR) exerted anti-MM effects without damaging normal hematopoietic cells. Thus, MMG49 CAR T cell therapy is promising for MM, and a receptor protein with a rare but physiologically relevant conformation can serve as a cancer immunotherapy target.

  • Subscribe to Nature Medicine for full access:

    $225

    Subscribe

Additional access options:

Already a subscriber?  Log in  now or  Register  for online access.

References

  1. 1.

    , & Antibody therapy of cancer. Nat. Rev. Cancer 12, 278–287 (2012).

  2. 2.

    , , , & Chimeric antigen receptor-modified T cells in chronic lymphoid leukemia. N. Engl. J. Med. 365, 725–733 (2011).

  3. 3.

    et al. Chimeric antigen receptor-modified T cells for acute lymphoid leukemia. N. Engl. J. Med. 368, 1509–1518 (2013).

  4. 4.

    et al. Mutational landscape and significance across 12 major cancer types. Nature 502, 333–339 (2013).

  5. 5.

    et al. Mutational heterogeneity in cancer and the search for new cancer-associated genes. Nature 499, 214–218 (2013).

  6. 6.

    & Tumour heterogeneity and cancer cell plasticity. Nature 501, 328–337 (2013).

  7. 7.

    et al. Engineered CAR T cells targeting the cancer-associated Tn-glycoform of the membrane mucin MUC1 control adenocarcinoma. Immunity 44, 1444–1454 (2016).

  8. 8.

    & Multiple myeloma. N. Engl. J. Med. 364, 1046–1060 (2011).

  9. 9.

    Treatment of multiple myeloma. Nat. Rev. Clin. Oncol. 8, 479–491 (2011).

  10. 10.

    , , & Current treatment landscape for relapsed and/or refractory multiple myeloma. Nat. Rev. Clin. Oncol. 12, 42–54 (2015).

  11. 11.

    et al. Selective expression of integrin alpha 4 beta 7 on a subset of human CD4+ memory T cells with hallmarks of gut-trophism. J. Immunol. 151, 717–729 (1993).

  12. 12.

    et al. Expression and function of the MAdCAM-1 receptor, integrin alpha 4 beta 7, on human leukocytes. J. Immunol. 153, 517–528 (1994).

  13. 13.

    , , & Global conformational rearrangements in integrin extracellular domains in outside-in and inside-out signaling. Cell 110, 599–511 (2002).

  14. 14.

    , & Conformational regulation of integrin structure and function. Annu. Rev. Biophys. Biomol. Struct. 31, 485–516 (2002).

  15. 15.

    et al. Structure-function analysis of the integrin beta 7 subunit: identification of domains involved in adhesion to MAdCAM-1. J. Immunol. 159, 1497–1505 (1997).

  16. 16.

    , & Bistable regulation of integrin adhesiveness by a bipolar metal ion cluster. Nat. Struct. Biol. 10, 995–1001 (2003).

  17. 17.

    et al. Identification, characterization, and epitope mapping of human monoclonal antibody J19 that specifically recognizes activated integrin α4β7. J. Biol. Chem. 287, 15749–15759 (2012).

  18. 18.

    et al. Topography of ligand-induced binding sites, including a novel cation-sensitive epitope (AP5) at the amino terminus, of the human integrin beta 3 subunit. J. Biol. Chem. 270, 11947–11954 (1995).

  19. 19.

    et al. Alpha 4 beta 7 integrin mediates lymphocyte binding to the mucosal vascular addressin MAdCAM-1. Cell 74, 185–195 (1993).

  20. 20.

    et al. Overexpression of c-maf is a frequent oncogenic event in multiple myeloma that promotes proliferation and pathological interactions with bone marrow stroma. Cancer Cell 5, 191–199 (2004).

  21. 21.

    et al. Integrin β7-mediated regulation of multiple myeloma cell adhesion, migration, and invasion. Blood 117, 6202–6213 (2011).

  22. 22.

    et al. Critical role for β7 integrins in formation of the gut-associated lymphoid tissue. Nature 382, 366–370 (1996).

  23. 23.

    et al. Nonmuscle myosin heavy chain IIA mediates integrin LFA-1 de-adhesion during T lymphocyte migration. J. Exp. Med. 205, 195–205 (2008).

  24. 24.

    et al. Case report of a serious adverse event following the administration of T cells transduced with a chimeric antigen receptor recognizing ERBB2. Mol. Ther. 18, 843–851 (2010).

  25. 25.

    , , , & Human T-lymphocyte cytotoxicity and proliferation directed by a single chimeric TCRζ/CD28 receptor. Nat. Biotechnol. 20, 70–75 (2002).

  26. 26.

    et al. Affinity-tuned ErbB2 or EGFR chimeric antigen receptor T cells exhibit an increased therapeutic index against tumors in mice. Cancer Res. 75, 3596–3607 (2015).

  27. 27.

    et al. The myeloma stem cell concept, revisited: from phenomenology to operational terms. Haematologica 101, 1451–1459 (2016).

  28. 28.

    et al. In multiple myeloma, clonotypic B lymphocytes are detectable among CD19+ peripheral blood cells expressing CD38, CD56, and monotypic Ig light chain. Blood 85, 436–447 (1995).

  29. 29.

    et al. NY-ESO-1–specific TCR–engineered T cells mediate sustained antigen-specific antitumor effects in myeloma. Nat. Med. 21, 914–921 (2015).

  30. 30.

    et al. Chimeric antigen receptor T cells against CD19 for multiple myeloma. N. Engl. J. Med. 373, 1040–1047 (2015).

  31. 31.

    et al. B-cell maturation antigen is a promising target for adoptive T-cell therapy of multiple myeloma. Clin. Cancer. Res. 19, 2048–2060 (2013).

  32. 32.

    et al. T cells expressing an anti-B-cell maturation antigen chimeric antigen receptor cause remissions of multiple myeloma. Blood 128, 1688–1700 (2016).

  33. 33.

    et al. Genetic modification of T cells redirected toward CS1 enhances eradication of myeloma cells. Clin. Cancer. Res. 20, 3989–4000 (2014).

  34. 34.

    Intracellular signalling controlling integrin activation in lymphocytes. Nat. Rev. Immunol. 5, 546–559 (2005).

  35. 35.

    et al. A monoclonal antibody recognizing human cancers with amplification/overexpression of the human epidermal growth factor receptor. Proc. Natl. Acad. Sci. USA 100, 639–644 (2003).

  36. 36.

    et al. Identification of the epitope for the epidermal growth factor receptor-specific monoclonal antibody 806 reveals that it preferentially recognizes an untethered form of the receptor. J. Biol. Chem. 279, 30375–30384 (2004).

  37. 37.

    et al. Differential EphA2 epitope display on normal versus malignant cells. Cancer Res. 63, 7907–7912 (2003).

  38. 38.

    et al. T cells redirected to EphA2 for the immunotherapy of glioblastoma. Mol. Ther. 21, 629–637 (2013).

  39. 39.

    , , & Targeting of a conformationally exposed, tumor-specific epitope of EGFR as a strategy for cancer therapy. Cancer Res. 72, 2924–2930 (2012).

  40. 40.

    et al. Antibodies specifically targeting a locally misfolded region of tumor associated EGFR. Proc. Natl. Acad. Sci. USA 106, 5082–5087 (2009).

  41. 41.

    , & Structural basis of integrin regulation and signaling. Annu. Rev. Immunol. 25, 619–647 (2007).

  42. 42.

    et al. CD40-activated human B cells: an alternative source of highly efficient antigen presenting cells to generate autologous antigen-specific T cells for adoptive immunotherapy. J. Clin. Invest. 100, 2757–2765 (1997).

  43. 43.

    et al. Efficient screening of retroviral cDNA expression libraries. Proc. Natl. Acad. Sci. USA 92, 9146–9150 (1995).

  44. 44.

    et al. Glycoproteins of 210,000 and 130,000 m.w. on activated T cells: cell distribution and antigenic relation to components on resting cells and T cell lines. J. Immunol. 132, 3011–3018 (1984).

  45. 45.

    , & C-terminal opening mimics 'inside-out' activation of integrin α5β1. Nat. Struct. Biol. 8, 412–416 (2001).

  46. 46.

    et al. Crystal structure of α5β1 integrin ectodomain: atomic details of the fibronectin receptor. J. Cell Biol. 197, 131–140 (2012).

  47. 47.

    et al. Multiplex genome engineering using CRISPR/Cas systems. Science 339, 819–823 (2013).

  48. 48.

    et al. CD28 costimulation provided through a CD19-specific chimeric antigen receptor enhances in vivo persistence and antitumor efficacy of adoptively transferred T cells. Cancer Res. 66, 10995–11004 (2006).

  49. 49.

    et al. Generation of CD19-chimeric antigen receptor modified CD8+ T cells derived from virus-specific central memory T cells. Blood 119, 72–82 (2012).

  50. 50.

    et al. CD48 as a novel molecular target for antibody therapy in multiple myeloma. Br. J. Haematol. 156, 213–224 (2012).

Download references

Acknowledgements

We thank the Kinki Cord Blood Bank for CB samples and T. Yamane (Osaka City General Hospital) and K. Koh (Osaka General Hospital for West Japan Railway Company) for MM samples. We also thank K. Terasaki, S. Ikeda, Y. Hayami, R. Inada, R. Urakawa, S. Hashiguchi, M. Iwai, and A. Kosugi for technical assistance, and Y. Kanakura (Osaka University), R. Burger (University of Kiel), I. Weissman (Stanford University), T. Kitamura (Tokyo University), and the NIH AIDS Reagents program for providing materials. This work was supported in part by a Grant-in-Aid for Scientific Research on Innovative Areas (Analysis and Synthesis of Multidimensional Immune Organ Network) from the Ministry of Education, Culture, Sports, Science and Technology of Japan (to J.T.); the Project for Development of Innovative Research on Cancer Therapeutics and Practical Research for Innovative Cancer Control from the Japan Agency for Medical Research and Development AMED (to N.H.); JSPS KAKENHI grant JP26461404 (to N.H.); and Japan Agency for Medical Research and Development–Core Research for Evolutional Science and Technology grant 15652237 (to A.K.).

Author information

Author notes

    • Yukiko Matsunaga

    Present address: Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario, Canada.

    • Yukiko Matsunaga
    •  & Kana Hasegawa

    These authors contributed equally to this work.

Affiliations

  1. Department of Cancer Stem Cell Biology, Osaka University Graduate School of Medicine, Osaka, Japan.

    • Naoki Hosen
    • , Kana Hasegawa
    • , Yuki Nakamura
    • , Mio Makita
    • , Kouki Watanabe
    • , Mikako Yoshida
    • , Kei Satoh
    •  & Yoshihiro Oka
  2. Department of Respiratory Medicine and Clinical Immunology, Osaka University Graduate School of Medicine, Osaka, Japan.

    • Naoki Hosen
    • , Sumiyuki Nishida
    • , Yoshihiro Oka
    •  & Atsushi Kumanogoh
  3. WPI Immunology Frontier Research Center, Osaka University, Osaka, Japan.

    • Naoki Hosen
    • , Yoshihiro Oka
    • , Kiyoshi Takeda
    •  & Atsushi Kumanogoh
  4. Laboratory of Protein Synthesis and Expression, Institute for Protein Research, Osaka University, Osaka, Japan.

    • Yukiko Matsunaga
    •  & Junichi Takagi
  5. Department of Gastroenterological Surgery, Osaka University Graduate School of Medicine, Osaka, Japan.

    • Hiroshi Matsuno
    •  & Junichi Nishimura
  6. Department of Cancer Immunotherapy, Osaka University Graduate School of Medicine, Osaka, Japan.

    • Soyoko Morimoto
    • , Jun Nakata
    •  & Akihiro Tsuboi
  7. Department of Cancer Immunology, Osaka University Graduate School of Medicine, Osaka, Japan.

    • Fumihiro Fujiki
    • , Hiroko Nakajima
    •  & Haruo Sugiyama
  8. Department of Hematology, Osaka General Hospital of West Japan Railway Company, Osaka, Japan.

    • Masahiro Manabe
  9. Department of Hematology, Fuchu Hospital, Osaka, Japan.

    • Hiroyoshi Ichihara
    • , Yasutaka Aoyama
    •  & Atsuko Mugitani
  10. Department of Hematology, Osaka City General Hospital, Osaka, Japan.

    • Takafumi Nakao
  11. Department of Hematology and Oncology, Osaka City University Graduate School of Medicine, Osaka, Japan.

    • Masayuki Hino
  12. Division of Immuno-Gene & Cell Therapy (Takara Bio), Jichi Medical University, Tochigi, Japan.

    • Ryosuke Uchibori
    •  & Keiya Ozawa
  13. Institute of Medical Science, University of Tokyo, Tokyo, Japan.

    • Keiya Ozawa
  14. Division of Immunology and Genome Biology, Medical Institute of Bioregulation, Kyushu University, Fukuoka, Japan.

    • Yoshihiro Baba
  15. Department of Hematology and Oncology, Nagoya University Graduate School of Medicine, Nagoya, Japan.

    • Seitaro Terakura
  16. Department of Pathology, Osaka University Graduate School of Medicine, Osaka, Japan.

    • Naoki Wada
    •  & Eiichi Morii
  17. Department of Microbiology and Immunology, Osaka University Graduate School of Medicine, Osaka, Japan.

    • Kiyoshi Takeda
  18. Japan Agency for Medical Research and Development–Core Research for Evolutional Science and Technology (AMED-CREST), Japan.

    • Kiyoshi Takeda
    •  & Atsushi Kumanogoh
  19. Department of Functional Diagnostic Science, Osaka University Graduate School of Medicine, Osaka, Japan.

    • Yusuke Oji

Authors

  1. Search for Naoki Hosen in:

  2. Search for Yukiko Matsunaga in:

  3. Search for Kana Hasegawa in:

  4. Search for Hiroshi Matsuno in:

  5. Search for Yuki Nakamura in:

  6. Search for Mio Makita in:

  7. Search for Kouki Watanabe in:

  8. Search for Mikako Yoshida in:

  9. Search for Kei Satoh in:

  10. Search for Soyoko Morimoto in:

  11. Search for Fumihiro Fujiki in:

  12. Search for Hiroko Nakajima in:

  13. Search for Jun Nakata in:

  14. Search for Sumiyuki Nishida in:

  15. Search for Akihiro Tsuboi in:

  16. Search for Yoshihiro Oka in:

  17. Search for Masahiro Manabe in:

  18. Search for Hiroyoshi Ichihara in:

  19. Search for Yasutaka Aoyama in:

  20. Search for Atsuko Mugitani in:

  21. Search for Takafumi Nakao in:

  22. Search for Masayuki Hino in:

  23. Search for Ryosuke Uchibori in:

  24. Search for Keiya Ozawa in:

  25. Search for Yoshihiro Baba in:

  26. Search for Seitaro Terakura in:

  27. Search for Naoki Wada in:

  28. Search for Eiichi Morii in:

  29. Search for Junichi Nishimura in:

  30. Search for Kiyoshi Takeda in:

  31. Search for Yusuke Oji in:

  32. Search for Haruo Sugiyama in:

  33. Search for Junichi Takagi in:

  34. Search for Atsushi Kumanogoh in:

Contributions

N.H. and J.T. designed the experiments; N.H., Y.M., K.H., H.M., Y.N., M. Makita., K.W., M.Y., K.S., S.M., F.F., H.N., J.Nakata., S.N., A.T., Y.Oka., R.U., K.O., Y.B., S.T., N.W., E.M., J.Nishimura., and Y.Oji. performed the experiments; M.Manabe., H.I., Y.A., A.M., T.N., and M.H. collected and analyzed clinical samples; N.H., Y.M., K.H., and J.T. analyzed the data; N.H., Y.M., K.H., K.T., H.S., J.T., and A.K. wrote the manuscript; and all authors reviewed and approved the final version of the manuscript.

Competing interests

N.H., H.S., J.T., and A.K. have applied for a Japanese patent entitled 'Antibody' through the Osaka University Office for University-Industry Collaboration. The other authors declare no competing financial interests.

Corresponding author

Correspondence to Naoki Hosen.

Supplementary information

PDF files

  1. 1.

    Supplementary Figures

    Supplementary Figures 1–15

  2. 2.

    Life Sciences Reporting Summary