Advertisement

Inhibiting IL-2 signaling and the regulatory T-cell pathway using computationally designed peptides

  • Tammy Price-Troska
  • Zhi-Zhang Yang
  • David Diller
  • Alexander Bayden
  • Mark Jarosinski
  • Joseph Audie
  • Stephen M. Ansell
PRECLINICAL STUDIES

Summary

Background Increased serum levels of soluble interleukin-2 (IL-2) receptor alpha (sIL-2Rα) are an indicator of poor prognosis in patients with B-cell non-Hodgkin lymphoma (NHL). By binding to IL-2, sIL-2Rα upregulates Foxp3 expression and induces the development of regulatory T (Treg) cells. Methods To inhibit the binding of IL-2 to sIL-2Rα with the goal of suppressing the induction of Foxp3 and decreasing Treg cell numbers, we developed peptides by structure-based computational design to disrupt the interaction between IL-2 and sIL-2Rα. Each peptide was screened using an enzyme-linked immunosorbent assay (ELISA), and 10 of 22 peptides showed variable capacity to inhibit IL-2/sIL-2Rα binding. Results We identified a lead candidate peptide, CMD178, which consistently reduced the expression of Foxp3 and STAT5 induced by IL-2/sIL-2Rα signaling. Furthermore, production of cytokines (IL-2/interferon gamma [IFN-γ]) and granules (perforin/granzyme B) was preserved in CD8+ T cells co-cultured with IL-2–stimulated CD4+ T cells that had been pretreated with CMD178 compared to CD8+ cells co-cultured with untreated IL-2–stimulated CD4+ T cells where it was inhibited. Conclusions We conclude that structure-based peptide design can be used to identify novel peptide inhibitors that block IL-2/sIL-2Rα signaling and inhibit Treg cell development. We anticipate that these peptides will have therapeutic potential in B-cell NHL and other malignancies.

Keywords

Computationally designed peptides Foxp3 Soluble IL-2Ra STAT5 Treg cells 

Notes

Acknowledgments

This work was supported in part by grants from the National Institutes of Health (P50 CA97274), the Leukemia & Lymphoma Society, the Landow Foundation, and the Predolin Foundation.

Compliance with ethical standards

Conflict of interest

All authors declare no conflict of interest.

Ethical approval

The authors declared that this article does not contain any studies with human or animal participants.

Informed consent

For this type of study, formal consent is not required.

References

  1. 1.
    Bien E, Balcerska A (2008) Serum soluble interleukin 2 receptor alpha in human cancer of adults and children: a review. Biomarkers 13(1):1–26CrossRefPubMedGoogle Scholar
  2. 2.
    Gupta M, Stenson M, O'Byrne M, Maurer MJ, Habermann T, Cerhan JR, Weiner GW, Witzig TE (2016) Comprehensive serum cytokine analysis identifies IL-1RA and soluble IL-2Ralpha as predictors of event-free survival in T-cell lymphoma. Ann Oncol: Off J Eur Soc for Med Oncol/ESMO 27(1):165–172CrossRefGoogle Scholar
  3. 3.
    Wang L, Liao DZ, Zhang J, Xia ZJ, Peng XW, Lu Y (2013) Clinical significance of serum soluble interleukin-2 receptor-alpha in extranodal natural killer/T-cell lymphoma (ENKTL): a predictive biomarker for treatment efficacy and valuable prognostic factor. Med Oncol 30(4):723CrossRefPubMedGoogle Scholar
  4. 4.
    Jo SA, Hwang SH, Chang CL, Kim SY, Shin HJ, Chung JS, Sol MY, Lee EY (2010) Clinical relevance of elevated levels of serum soluble interleukin-2 receptor alpha (sIL-2Ralpha) in patients with non-Hodgkin's lymphoma. Korean J Lab Med 30(6):600–605CrossRefPubMedGoogle Scholar
  5. 5.
    Jeffery HC, Jeffery LE, Lutz P, Corrigan M, Webb GJ, Hirschfield GM, Adams DH, Oo YH (2017) Low-dose interleukin-2 promotes STAT-5 phosphorylation, Treg survival and CTLA-4-dependent function in autoimmune liver diseases. Clin Exp Immunol 188(3):394–411CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Liu W, Putnam AL, Xu-Yu Z, Szot GL, Lee MR, Zhu S, Gottlieb PA, Kapranov P, Gingeras TR, Fazekas de St Groth B, Clayberger C, Soper DM, Ziegler SF, Bluestone JA (2006) CD127 expression inversely correlates with FoxP3 and suppressive function of human CD4+ T reg cells. J Exp Med 203(7):1701–1711CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Mahmud SA, Manlove LS, Farrar MA (2013) Interleukin-2 and STAT5 in regulatory T cell development and function. JAKSTAT 2(1):e23154PubMedPubMedCentralGoogle Scholar
  8. 8.
    Ai WZ, Hou JZ, Zeiser R, Czerwinski D, Negrin RS, Levy R (2009) Follicular lymphoma B cells induce the conversion of conventional CD4+ T cells to T-regulatory cells. Int J Cancer 124(1):239–244CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Binder M, O'Byrne MM, Maurer MJ, Ansell S, Feldman AL, Cerhan J, Novak A, Porrata LF, Markovic S, Link BK, Witzig TE (2017) Associations between elevated pre-treatment serum cytokines and peripheral blood cellular markers of immunosuppression in patients with lymphoma. Am J Hematol 92(8):752–758CrossRefPubMedGoogle Scholar
  10. 10.
    Mir MA, Maurer MJ, Ziesmer SC, Slager SL, Habermann T, Macon WR, Link BK, Syrbu S, Witzig T, Friedberg JW, Press O, LeBlanc M, Cerhan JR, Novak A, Ansell SM (2015) Elevated serum levels of IL-2R, IL-1RA, and CXCL9 are associated with a poor prognosis in follicular lymphoma. Blood 125(6):992–998CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Zorn E, Nelson EA, Mohseni M, Porcheray F, Kim H, Litsa D, Bellucci R, Raderschall E, Canning C, Soiffer RJ, Frank DA, Ritz J (2006) IL-2 regulates FOXP3 expression in human CD4+CD25+ regulatory T cells through a STAT-dependent mechanism and induces the expansion of these cells in vivo. Blood 108(5):1571–1579CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Yang ZZ, Grote DM, Ziesmer SC, Manske MK, Witzig TE, Novak AJ, Ansell SM (2011) Soluble IL-2Ralpha facilitates IL-2-mediated immune responses and predicts reduced survival in follicular B-cell non-Hodgkin lymphoma. Blood 118(10):2809–2820CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Janas ML, Groves P, Kienzle N, Kelso A (2005) IL-2 regulates perforin and granzyme gene expression in CD8+ T cells independently of its effects on survival and proliferation. J Immunol 175(12):8003–8010CrossRefPubMedGoogle Scholar
  14. 14.
    Onoprienko LV, Mikhaleva II, Lunev VE, Nesmeianov VA, Ivanov VT (1989) Synthesis and immunochemical properties of peptides corresponding to sequences 59-72 and 25-36 of human interleukin-2. Bioorg Khim 15(7):908–921PubMedGoogle Scholar
  15. 15.
    Wang X, Rickert M, Garcia KC (2005) Structure of the quaternary complex of interleukin-2 with its alpha, beta, and gammac receptors. Science 310(5751):1159–1163CrossRefPubMedGoogle Scholar
  16. 16.
    Du J, Yang H, Zhang D, Wang J, Guo H, Peng B, Guo Y, Ding J (2010) Structural basis for the blockage of IL-2 signaling by therapeutic antibody basiliximab. J Immunol 184(3):1361–1368CrossRefPubMedGoogle Scholar
  17. 17.
    Boyman O, Sprent J (2012) The role of interleukin-2 during homeostasis and activation of the immune system. Nat Rev Immunol 12(3):180–190CrossRefPubMedGoogle Scholar
  18. 18.
    Liao W, Lin JX, Leonard WJ (2013) Interleukin-2 at the crossroads of effector responses, tolerance, and immunotherapy. Immunity 38(1):13–25CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Emerson SD, Palermo R, Liu CM, Tilley JW, Chen L, Danho W, Madison VS, Greeley DN, Ju G, Fry DC (2003) NMR characterization of interleukin-2 in complexes with the IL-2Ralpha receptor component, and with low molecular weight compounds that inhibit the IL-2/IL-Ralpha interaction. Protein Sci 12(4):811–822CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Sauve K, Nachman M, Spence C, Bailon P, Campbell E, Tsien WH, Kondas JA, Hakimi J, Localization JG (1991) In human interleukin 2 of the binding site to the alpha chain (p55) of the interleukin 2 receptor. Proc Natl Acad Sci U S A 88(11):4636–4640CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Thanos CD, DeLano WL, Wells JA (2006) Hot-spot mimicry of a cytokine receptor by a small molecule. Proc Natl Acad Sci U S A 103(42):15422–15427CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Rickert M, Wang X, Boulanger MJ, Goriatcheva N, Garcia KC (2005) The structure of interleukin-2 complexed with its alpha receptor. Science 308(5727):1477–1480CrossRefPubMedGoogle Scholar
  23. 23.
    Liu BY, Zhu P, Luo HB, Fu N (2006) screening of short peptides binding to cell surface interleukin-2 receptor alpha chain. Nan Fang Yi Ke Da Xue Xue Bao 26(7):971–974PubMedGoogle Scholar
  24. 24.
    Tzankov A, Meier C, Hirschmann P, Went P, Pileri SA, Dirnhofer S (2008) Correlation of high numbers of intratumoral FOXP3+ regulatory T cells with improved survival in germinal center-like diffuse large B-cell lymphoma, follicular lymphoma and classical Hodgkin's lymphoma. Haematologica 93(2):193–200CrossRefPubMedGoogle Scholar
  25. 25.
    Hude I, Sasse S, Engert A, Brockelmann PJ (2017) The emerging role of immune checkpoint inhibition in malignant lymphoma. Haematologica 102(1):30–42CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Tammy Price-Troska
    • 1
  • Zhi-Zhang Yang
    • 1
  • David Diller
    • 2
    • 3
  • Alexander Bayden
    • 2
    • 4
  • Mark Jarosinski
    • 2
  • Joseph Audie
    • 2
    • 5
  • Stephen M. Ansell
    • 1
  1. 1.Division of Hematology, Department of MedicineMayo ClinicRochesterUSA
  2. 2.CMD Bioscience, Inc.New HavenUSA
  3. 3.Data2 Discovery ConsultingEast WindsorUSA
  4. 4.IDEAYA Biosciences, Inc.CaliforniaUSA
  5. 5.Sacred Heart UniversityFairfieldUSA

Personalised recommendations