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siRNA-mediated silencing of PD-1 ligands enhances tumor-specific human T-cell effector functions

Gene Therapy volume 19, pages 959–966 (2012)Cite this article

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Abstract

Adoptive cell therapy using tumor-specific T cells is a promising strategy for treating patients with malignancy. However, accumulating evidences have demonstrated that optimal function of tumor-reactive T cells is often attenuated by negative regulatory signal(s) delivered through receptors, such as cytotoxic T-lymphocyte antigen 4 (CTLA-4), programmed death 1 (PD-1), and their cognate ligands. Although systemic blocking of these molecules needs careful attention on the risk of uncontrolled immune activation, selective inhibition of negative signals in tumor-specific T cells by their genetic modification is an attractive approach to overcome immunological suppression in cancer patients. Here, we demonstrate the improved effector functions of tumor-specific CD4+ and CD8+ human T cells by small interfering RNA (siRNA) -mediated silencing of PD-1 ligands, PD-L1 or PD-L2. Tumor antigen MAGE-A4-specific human T-cell clones upregulated the expression of PD-1 ligands upon activation. siRNA-mediated knockdown of PD-L1 or -L2 enhanced the interferon-γ production and antigen-specific cytotoxicity of these cells. Peripheral blood mononuclear cells transduced with a retroviral vector encoding MAGE-A4-specific T-cell receptor α/β chains also increased their effector functions by this modification. These results suggest that siRNA-mediated knockdown of PD-1 ligands is an attractive strategy to inhibit a negative regulatory mechanism of tumor-specific T cells resulting in enhanced efficacy of adoptive T-cell therapy of cancer using genetically modified autologous lymphocytes.

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References

  1. June CH . Adoptive T cell therapy for cancer in the clinic. J Clin Invest 2007; 117: 1466–1476.

    Article  CAS  Google Scholar 

  2. Zitvogel L, Tesniere A, Kroemer G . Cancer despite immunosurveillance: immunoselection and immunosubversion. Nat Rev Immunol 2006; 6: 715–727.

    Article  CAS  Google Scholar 

  3. Gattinoni L, Powell Jr DJ, Rosenberg SA, Restifo NP . Adoptive immunotherapy for cancer: building on success. Nat Rev Immunol 2006; 6: 383–393.

    Article  CAS  Google Scholar 

  4. Stephan MT, Ponomarev V, Brentjens RJ, Chang AH, Dobrenkov KV, Heller G et al. T cell-encoded CD80 and 4-1BBL induce auto- and transcostimulation, resulting in potent tumor rejection. Nat Med 2007; 13: 1440–1449.

    Article  CAS  Google Scholar 

  5. Greenwald RJ, Latchman YE, Sharpe AH . Negative co-receptors on lymphocytes. Curr Opin Immunol 2002; 14: 391–396.

    Article  CAS  Google Scholar 

  6. Okazaki T, Iwai Y, Honjo T . New regulatory co-receptors: inducible co-stimulator and PD-1. Curr Opin Immunol 2002; 14: 779–782.

    Article  CAS  Google Scholar 

  7. Zou W, Chen L . Inhibitory B7-family molecules in the tumour microenvironment. Nat Rev Immunol 2008; 8: 467–477.

    Article  CAS  Google Scholar 

  8. Keir ME, Butte MJ, Freeman GJ, Sharpe AH . PD-1 and its ligands in tolerance and immunity. Annu Rev Immunol 2008; 26: 677–704.

    Article  CAS  Google Scholar 

  9. Brown JA, Dorfman DM, Ma FR, Sullivan EL, Munoz O, Wood CR et al. Blockade of programmed death-1 ligands on dendritic cells enhances T cell activation and cytokine production. J Immunol 2003; 170: 1257–1266.

    Article  CAS  Google Scholar 

  10. Dong H, Strome SE, Matteson EL, Moder KG, Flies DB, Zhu G et al. Costimulating aberrant T cell responses by B7-H1 autoantibodies in rheumatoid arthritis. J Clin Invest 2003; 111: 363–370.

    Article  CAS  Google Scholar 

  11. Bennett F, Luxenberg D, Ling V, Wang IM, Marquette K, Lowe D et al. Program death-1 engagement upon TCR activation has distinct effects on costimulation and cytokine-driven proliferation: attenuation of ICOS, IL-4, and IL-21, but not CD28, IL-7, and IL-15 responses. J Immunol 2003; 170: 711–718.

    Article  CAS  Google Scholar 

  12. Kuipers H, Muskens F, Willart M, Hijdra D, van Assema FB, Coyle AJ et al. Contribution of the PD-1 ligands/PD-1 signaling pathway to dendritic cell-mediated CD4+ T cell activation. Eur J Immunol 2006; 36: 2472–2482.

    Article  CAS  Google Scholar 

  13. Kim YS, Park GB, Lee HK, Song H, Choi IH, Lee WJ et al. Cross-linking of B7-H1 on EBV-transformed B cells induces apoptosis through reactive oxygen species production, JNK signaling activation, and fasL expression. J Immunol 2008; 181: 6158–6169.

    Article  CAS  Google Scholar 

  14. Dembic Z, Haas W, Weiss S, McCubrey J, Kiefer H, von Boehmer H et al. Transfer of specificity by murine alpha and beta T-cell receptor genes. Nature 1986; 320: 232–238.

    Article  CAS  Google Scholar 

  15. Schmitt TM, Ragnarsson GB, Greenberg PD . T cell receptor gene therapy for cancer. Hum Gene Ther 2009; 20: 1240–1248.

    Article  CAS  Google Scholar 

  16. Larin SS, Georgiev GP, Kiselev SL . Gene transfer approaches in cancer immunotherapy. Gene Therapy 2004; 11 (Suppl 1): S18–S25.

    Article  CAS  Google Scholar 

  17. Rosenberg SA, Restifo NP, Yang JC, Morgan RA, Dudley ME . Adoptive cell transfer: a clinical path to effective cancer immunotherapy. Nat Rev Cancer 2008; 8: 299–308.

    Article  CAS  Google Scholar 

  18. Miyahara Y, Naota H, Wang L, Hiasa A, Goto M, Watanabe M et al. Determination of cellularly processed HLA-A2402-restricted novel CTL epitopes derived from two cancer germ line genes, MAGE-A4 and SAGE. Clin Cancer Res 2005; 11: 5581–5589.

    Article  CAS  Google Scholar 

  19. Hiasa A, Hirayama M, Nishikawa H, Kitano S, Nukaya I, Yu SS et al. Long-term phenotypic, functional and genetic stability of cancer-specific T-cell receptor (TCR) alphabeta genes transduced to CD8+ T cells. Gene Therapy 2008; 15: 695–699.

    Article  CAS  Google Scholar 

  20. Okamoto S, Mineno J, Ikeda H, Fujiwara H, Yasukawa M, Shiku H et al. Improved expression and reactivity of transduced tumor-specific TCRs in human lymphocytes by specific silencing of endogenous TCR. Cancer Res 2009; 69: 9003–9011.

    Article  CAS  Google Scholar 

  21. Roy M, Aruffo A, Ledbetter J, Linsley P, Kehry M, Noelle R . Studies on the interdependence of gp39 and B7 expression and function during antigen-specific immune responses. Eur J Immunol 1995; 25: 596–603.

    Article  CAS  Google Scholar 

  22. Kato T, Hakamada R, Yamane H, Nariuchi H . Induction of IL-12 p40 messenger RNA expression and IL-12 production of macrophages via CD40-CD40 ligand interaction. J Immunol 1996; 156: 3932–3938.

    CAS  PubMed  Google Scholar 

  23. Butte MJ, Keir ME, Phamduy TB, Sharpe AH, Freeman GJ . Programmed death-1 ligand 1 interacts specifically with the B7-1 costimulatory molecule to inhibit T cell responses. Immunity 2007; 27: 111–122.

    Article  CAS  Google Scholar 

  24. Mirza N, Duque MA, Dominguez AL, Schrum AG, Dong H, Lustgarten J . B7-H1 expression on old CD8+ T cells negatively regulates the activation of immune responses in aged animals. J Immunol 2010; 184: 5466–5474.

    Article  CAS  Google Scholar 

  25. Messal N, Serriari NE, Pastor S, Nunes JA, Olive D . PD-L2 is expressed on activated human T cells and regulates their function. Mol Immunol 2011; 48: 2214–2219.

    Article  CAS  Google Scholar 

  26. Nourry C, Grant SG, Borg JP . PDZ domain proteins: plug and play! Sci STKE 2003; 2003: RE7.

    PubMed  Google Scholar 

  27. Borkner L, Kaiser A, van de Kasteele W, Andreesen R, Mackensen A, Haanen JB et al. RNA interference targeting programmed death receptor-1 improves immune functions of tumor-specific T cells. Cancer Immunol Immunother 2010; 59: 1173–1183.

    Article  CAS  Google Scholar 

  28. Greenwald RJ, Freeman GJ, Sharpe AH . The B7 family revisited. Annu Rev Immunol 2005; 23: 515–548.

    Article  Google Scholar 

  29. Liu Y, Yu Y, Yang S, Zeng B, Zhang Z, Jiao G et al. Regulation of arginase I activity and expression by both PD-1 and CTLA-4 on the myeloid-derived suppressor cells. Cancer Immunol Immunother 2009; 58: 687–697.

    Article  CAS  Google Scholar 

  30. Poschke I, Mougiakakos D, Hansson J, Masucci GV, Kiessling R . Immature immunosuppressive CD14+HLA-DR-/low cells in melanoma patients are Stat3hi and overexpress CD80, CD83, and DC-sign. Cancer Res 2010; 70: 4335–4345.

    Article  CAS  Google Scholar 

  31. Liu X, Gao JX, Wen J, Yin L, Li O, Zuo T et al. B7DC/PDL2 promotes tumor immunity by a PD-1-independent mechanism. J Exp Med 2003; 197: 1721–1730.

    Article  CAS  Google Scholar 

  32. Wang S, Bajorath J, Flies DB, Dong H, Honjo T, Chen L . Molecular modeling and functional mapping of B7-H1 and B7-DC uncouple costimulatory function from PD-1 interaction. J Exp Med 2003; 197: 1083–1091.

    Article  CAS  Google Scholar 

  33. Johnson LA, Morgan RA, Dudley ME, Cassard L, Yang JC, Hughes MS et al. Gene therapy with human and mouse T-cell receptors mediates cancer regression and targets normal tissues expressing cognate antigen. Blood 2009; 114: 535–546.

    Article  CAS  Google Scholar 

  34. Finn OJ . Cancer immunology. N Engl J Med 2008; 358: 2704–2715.

    Article  CAS  Google Scholar 

  35. Blank C, Mackensen A . Contribution of the PD-L1/PD-1 pathway to T-cell exhaustion: an update on implications for chronic infections and tumor evasion. Cancer Immunol Immunother 2007; 56: 739–745.

    Article  Google Scholar 

  36. Ostrand-Rosenberg S, Sinha P . Myeloid-derived suppressor cells: linking inflammation and cancer. J Immunol 2009; 182: 4499–4506.

    Article  CAS  Google Scholar 

  37. Bendle GM, Linnemann C, Hooijkaas AI, Bies L, de Witte MA, Jorritsma A et al. Lethal graft-versus-host disease in mouse models of T cell receptor gene therapy. Nat Med 2010; 16: 565–570.

    Article  CAS  Google Scholar 

  38. Naota H, Miyahara Y, Okumura S, Kuzushima K, Akatsuka Y, Hiasa A et al. Generation of peptide-specific CD8+ T cells by phytohemagglutinin-stimulated antigen-mRNA-transduced CD4+ T cells. J Immunol Methods 2006; 314: 54–66.

    Article  CAS  Google Scholar 

  39. Nishikawa H, Tanida K, Ikeda H, Sakakura M, Miyahara Y, Aota T et al. Role of SEREX-defined immunogenic wild-type cellular molecules in the development of tumor-specific immunity. Proc Natl Acad Sci USA 2001; 98: 14571–14576.

    Article  CAS  Google Scholar 

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Acknowledgements

We gratefully acknowledge the skilled technical assistance of Chisaki Hyuga. This work was supported partly by Grants-in-Aid for Scientific Research on Priority Areas from the Ministry of Education, Culture, Sports, Science and Technology of Japan to HS.

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Authors and Affiliations

  1. Department of Immuno-Gene Therapy, Mie University Graduate School of Medicine, Mie, Japan

    K Iwamura, H Naota, H Ikeda & H Shiku

  2. Department of Cellular and Molecular Immunology, Mie University Graduate School of Medicine, Mie, Japan

    T Kato

  3. Department of Cancer Vaccine, Mie University Graduate School of Medicine, Mie, Japan

    Y Miyahara & H Shiku

  4. Center for Cell and Gene Therapy, Takara Bio Inc., Shiga, Japan

    J Mineno

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  1. K Iwamura
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Correspondence to H Shiku.

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Iwamura, K., Kato, T., Miyahara, Y. et al. siRNA-mediated silencing of PD-1 ligands enhances tumor-specific human T-cell effector functions. Gene Ther 19, 959–966 (2012). https://doi.org/10.1038/gt.2011.185

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