Advertisement

Cellular and Molecular Life Sciences

, Volume 75, Issue 22, pp 4223–4234 | Cite as

Inhibition of SRC family kinases facilitates anti-CTLA4 immunotherapy in head and neck squamous cell carcinoma

  • Guang-Tao Yu
  • Liang Mao
  • Lei Wu
  • Wei-Wei Deng
  • Lin-Lin Bu
  • Jian-Feng Liu
  • Lei Chen
  • Lei-Lei Yang
  • Hao Wu
  • Wen-Feng Zhang
  • Zhi-Jun Sun
Original Article

Abstract

The immune system plays a critical role in the establishment, development, and progression of head and neck squamous cell carcinoma (HNSCC). As treatment with single-immune checkpoint agent results in a lower response rate in patients, it is important to investigate new strategies to maintain favorable anti-tumor immune response. Herein, the combination immunotherapeutic value of CTLA4 blockade and SFKs inhibition was assessed in transgenic HNSCC mouse model. Our present work showed that tumor growth was not entirely controlled when HNSCC model mice were administered anti-CTLA4 chemotherapeutic treatment. Moreover, it was observed that Src family kinases (SFKs) were hyper-activated and lack of anti-tumor immune responses following anti-CTLA4 chemotherapeutic treatment. We hypothesized that activation of SFKs is a mechanism of anti-CTLA4 immunotherapy resistance. We, therefore, carried out combined drug therapy using anti-CTLA4 mAbs and an SFKs’ inhibitor, dasatinib. As expected, dasatinib and anti-CTLA4 synergistically inhibited tumor growth in Tgfbr1/Pten 2cKO mice. Furthermore, dasatinib and anti-CTLA4 combined to reduce the number of myeloid-derived suppressor cells and Tregs, increasing the CD8+ T cell-to-Tregs ratio. We also found that combining dasatinib with anti-CTLA4 therapy significantly attenuated the expression of p-STAT3Y705 and Ki67 in tumoral environment. These results suggest that combination therapy with SFKs inhibitors may be a useful therapeutic approach to increase the efficacy of anti-CTLA4 immunotherapy in HNSCC.

Keywords

CTLA4 Dasatinib MDSCs Tregs Immunotherapy HNSCC 

Abbreviations

aCTLA4

Anti-CTLA4

CTLA4

Cytotoxic T-lymphocyte-associated antigen 4

Dys

Dysplasia

HNSCC

Head and neck squamous cell carcinoma

HPV

Human papillomavirus

LN

Lymph node

MDSCs

Myeloid-derived suppressor cells

SFKs

Src family kinases

TAMs

Tumor-associated macrophages

TIL

Tumor infiltrate lymphocytes

Notes

Acknowledgements

We thank Zhi-Yong Chen and Dong Chen for excellent technical support. And also thank Wuhan Institute of Biotechnology for their Public Technology Service Platform. Meanwhile, this study was supported by National Natural Science Foundation of China (NFSC): 81672668, 81472528, and 81472529, and the Fundamental Research Funds for the Central Universities (2042017kf0171).

Compliance with ethical standards

Ethical standards

Animal studies were approved and supervised by the Animal Care and Use Committee of Wuhan University. The ethical approval number is 2014C66.

Conflict of interest

The authors declare that they have no competing interests.

Supplementary material

18_2018_2863_MOESM1_ESM.doc (12.5 mb)
Supplementary material 1 (DOC 12783 kb)

References

  1. 1.
    Bose P, Brockton NT, Dort JC (2013) Head and neck cancer: from anatomy to biology. Int J Cancer 133:2013–2023CrossRefGoogle Scholar
  2. 2.
    Siegel RL, Miller KD, Jemal A (2017) Cancer statistics, 2017. CA Cancer J Clin 67:7–30CrossRefGoogle Scholar
  3. 3.
    Ferris RL (2015) Immunology and immunotherapy of head and neck cancer. J Clin Oncol 33:3293–3304CrossRefGoogle Scholar
  4. 4.
    Jie HB, Schuler PJ, Lee SC, Srivastava RM, Argiris A, Ferrone S, Whiteside TL, Ferris RL (2015) CTLA-4(+) regulatory T cells increased in cetuximab-treated head and neck cancer patients suppress NK cell cytotoxicity and correlate with poor prognosis. Cancer Res 75:2200–2210CrossRefGoogle Scholar
  5. 5.
    Weed DT, Vella JL, Reis IM, De la Fuente AC, Gomez C, Sargi Z, Nazarian R, Califano J, Borrello I, Serafini P (2015) Tadalafil reduces myeloid-derived suppressor cells and regulatory T cells and promotes tumor immunity in patients with head and neck squamous cell carcinoma. Clin Cancer Res 21:39–48CrossRefGoogle Scholar
  6. 6.
    Moy JD, Moskovitz JM, Ferris RL (2017) Biological mechanisms of immune escape and implications for immunotherapy in head and neck squamous cell carcinoma. Eur J Cancer 76:152–166CrossRefGoogle Scholar
  7. 7.
    Economopoulou P, Agelaki S, Perisanidis C, Giotakis EI, Psyrri A (2016) The promise of immunotherapy in head and neck squamous cell carcinoma. Ann Oncol 27:1675–1685CrossRefGoogle Scholar
  8. 8.
    Baumeister SH, Freeman GJ, Dranoff G, Sharpe AH (2016) Coinhibitory pathways in immunotherapy for cancer. Annu Rev Immunol 34:539–573CrossRefGoogle Scholar
  9. 9.
    Yu GT, Bu LL, Zhao YY, Mao L, Deng WW, Wu TF, Zhang WF, Sun ZJ (2016) CTLA4 blockade reduces immature myeloid cells in head and neck squamous cell carcinoma. Oncoimmunology 5:e1151594CrossRefGoogle Scholar
  10. 10.
    Patel A, Sabbineni H, Clarke A, Somanath PR (2016) Novel roles of Src in cancer cell epithelial-to-mesenchymal transition, vascular permeability, microinvasion and metastasis. Life Sci 157:52–61CrossRefGoogle Scholar
  11. 11.
    Montero JC, Seoane S, Ocana A, Pandiella A (2011) Inhibition of SRC family kinases and receptor tyrosine kinases by dasatinib: possible combinations in solid tumors. Clin Cancer Res 17:5546–5552CrossRefGoogle Scholar
  12. 12.
    Baselga J, Cervantes A, Martinelli E, Chirivella I, Hoekman K, Hurwitz HI, Jodrell DI, Hamberg P, Casado E, Elvin P et al (2010) Phase I safety, pharmacokinetics, and inhibition of SRC activity study of saracatinib in patients with solid tumors. Clin Cancer Res 16:4876–4883CrossRefGoogle Scholar
  13. 13.
    Herold CI, Chadaram V, Peterson BL, Marcom PK, Hopkins J, Kimmick GG, Favaro J, Hamilton E, Welch RA, Bacus S, Blackwell KL (2011) Phase II trial of dasatinib in patients with metastatic breast cancer using real-time pharmacodynamic tissue biomarkers of Src inhibition to escalate dosing. Clin Cancer Res 17:6061–6070CrossRefGoogle Scholar
  14. 14.
    Bian Y, Hall B, Sun ZJ, Molinolo A, Chen W, Gutkind JS, Waes CV, Kulkarni AB (2012) Loss of TGF-beta signaling and PTEN promotes head and neck squamous cell carcinoma through cellular senescence evasion and cancer-related inflammation. Oncogene 31:3322–3332CrossRefGoogle Scholar
  15. 15.
    Sun ZJ, Zhang L, Hall B, Bian Y, Gutkind JS, Kulkarni AB (2012) Chemopreventive and chemotherapeutic actions of mTOR inhibitor in genetically defined head and neck squamous cell carcinoma mouse model. Clin Cancer Res 18:5304–5313CrossRefGoogle Scholar
  16. 16.
    Wolchok JD, Saenger Y (2008) The mechanism of anti-CTLA-4 activity and the negative regulation of T-cell activation. Oncologist 13(Suppl 4):2–9CrossRefGoogle Scholar
  17. 17.
    Wen X, Ding Y, Li J, Zhao J, Peng R, Li D, Zhu B, Wang Y, Zhang X, Zhang X (2017) The experience of immune checkpoint inhibitors in Chinese patients with metastatic melanoma: a retrospective case series. Cancer Immunol Immunother 66:1153–1162CrossRefGoogle Scholar
  18. 18.
    Cabel L, Loir E, Gravis G, Lavaud P, Massard C, Albiges L, Baciarello G, Loriot Y, Fizazi K (2017) Long-term complete remission with ipilimumab in metastatic castrate-resistant prostate cancer: case report of two patients. J Immunother Cancer 5:31CrossRefGoogle Scholar
  19. 19.
    Bagley SJ, Kothari S, Aggarwal C, Bauml JM, Alley EW, Evans TL, Kosteva JA, Ciunci CA, Gabriel PE, Thompson JC et al (2017) Pretreatment neutrophil-to-lymphocyte ratio as a marker of outcomes in nivolumab-treated patients with advanced non-small-cell lung cancer. Lung Cancer 106:1–7CrossRefGoogle Scholar
  20. 20.
    Schadendorf D, Hodi FS, Robert C, Weber JS, Margolin K, Hamid O, Patt D, Chen TT, Berman DM, Wolchok JD (2015) Pooled analysis of long-term survival data from phase II and phase III Trials of ipilimumab in unresectable or metastatic melanoma. J Clin Oncol 33:1889–1894CrossRefGoogle Scholar
  21. 21.
    Sharma P, Hu-Lieskovan S, Wargo JA, Ribas A (2017) Primary, adaptive, and acquired resistance to cancer immunotherapy. Cell 168:707–723CrossRefGoogle Scholar
  22. 22.
    Gao J, Shi LZ, Zhao H, Chen J, Xiong L, He Q, Chen T, Roszik J, Bernatchez C, Woodman SE et al (2016) Loss of IFN-gamma pathway genes in tumor cells as a mechanism of resistance to anti-CTLA-4 therapy. Cell 167(397–404):e9Google Scholar
  23. 23.
    Peng W, Chen JQ, Liu C, Malu S, Creasy C, Tetzlaff MT, Xu C, McKenzie JA, Zhang C, Liang X et al (2016) Loss of PTEN promotes resistance to T cell-mediated immunotherapy. Cancer Discov 6:202–216CrossRefGoogle Scholar
  24. 24.
    Guarino M (2010) Src signaling in cancer invasion. J Cell Physiol 223:14–26PubMedGoogle Scholar
  25. 25.
    Kreutzman A, Ilander M, Porkka K, Vakkila J, Mustjoki S (2014) Dasatinib promotes Th1-type responses in granzyme B expressing T-cells. Oncoimmunology 3:e28925CrossRefGoogle Scholar
  26. 26.
    Okamoto W, Okamoto I, Yoshida T, Okamoto K, Takezawa K, Hatashita E, Yamada Y, Kuwata K, Arao T, Yanagihara K et al (2010) Identification of c-Src as a potential therapeutic target for gastric cancer and of MET activation as a cause of resistance to c-Src inhibition. Mol Cancer Ther 9:1188–1197CrossRefGoogle Scholar
  27. 27.
    Nagaraj NS, Smith JJ, Revetta F, Washington MK, Merchant NB (2010) Targeted inhibition of SRC kinase signaling attenuates pancreatic tumorigenesis. Mol Cancer Ther 9:2322–2332CrossRefGoogle Scholar
  28. 28.
    Johnson FM, Saigal B, Talpaz M, Donato NJ (2005) Dasatinib (BMS-354825) tyrosine kinase inhibitor suppresses invasion and induces cell cycle arrest and apoptosis of head and neck squamous cell carcinoma and non-small cell lung cancer cells. Clin Cancer Res 11:6924–6932CrossRefGoogle Scholar
  29. 29.
    Mao L, Deng WW, Yu GT, Bu LL, Liu JF, Ma SR, Wu L, Kulkarni AB, Zhang WF, Sun ZJ (2017) Inhibition of SRC family kinases reduces myeloid-derived suppressor cells in head and neck cancer. Int J Cancer 140:1173–1185CrossRefGoogle Scholar
  30. 30.
    Kim LC, Song L, Haura EB (2009) Src kinases as therapeutic targets for cancer. Nat Rev Clin Oncol 6:587–595CrossRefGoogle Scholar
  31. 31.
    Verhagen AM, Wallace ME, Goradia A, Jones SA, Croom HA, Metcalf D, Collinge JE, Maxwell MJ, Hibbs ML, Alexander WS et al (2009) A kinase-dead allele of Lyn attenuates autoimmune disease normally associated with Lyn deficiency. J Immunol 182:2020–2029CrossRefGoogle Scholar
  32. 32.
    Hochgrafe F, Zhang L, O’Toole SA, Browne BC, Pinese M, Porta Cubas A, Lehrbach GM, Croucher DR, Rickwood D, Boulghourjian A et al (2010) Tyrosine phosphorylation profiling reveals the signaling network characteristics of Basal breast cancer cells. Cancer Res 70:9391–9401CrossRefGoogle Scholar
  33. 33.
    Maccalli C, Parmiani G, Ferrone S (2017) Immunomodulating and immunoresistance properties of cancer-initiating cells: implications for the clinical success of immunotherapy. Immunol Investig 46:221–238CrossRefGoogle Scholar
  34. 34.
    Yu GT, Bu LL, Huang CF, Zhang WF, Chen WJ, Gutkind JS, Kulkarni AB, Sun ZJ (2015) PD-1 blockade attenuates immunosuppressive myeloid cells due to inhibition of CD47/SIRPalpha axis in HPV negative head and neck squamous cell carcinoma. Oncotarget 6:42067–42080PubMedPubMedCentralGoogle Scholar
  35. 35.
    Linehan DC, Goedegebuure PS (2005) CD25+CD4+ regulatory T-cells in cancer. Immunol Res 32:155–168CrossRefGoogle Scholar
  36. 36.
    Qian L, Liu Y, Wang S, Gong W, Jia X, Liu L, Ye F, Ding J, Xu Y, Fu Y, Tian F (2017) NKG2D ligand RAE1epsilon induces generation and enhances the inhibitor function of myeloid-derived suppressor cells in mice. J Cell Mol Med 21:2046–2054CrossRefGoogle Scholar
  37. 37.
    Quezada SA, Peggs KS, Curran MA, Allison JP (2006) CTLA4 blockade and GM-CSF combination immunotherapy alters the intratumor balance of effector and regulatory T cells. J Clin Investig 116:1935–1945CrossRefGoogle Scholar
  38. 38.
    Boroughs LK, DeBerardinis RJ (2015) Metabolic pathways promoting cancer cell survival and growth. Nat Cell Biol 17:351–359CrossRefGoogle Scholar
  39. 39.
    Banerjee K, Resat H (2016) Constitutive activation of STAT3 in breast cancer cells: a review. Int J Cancer 138:2570–2578CrossRefGoogle Scholar

Copyright information

© Springer International Publishing AG, part of Springer Nature 2018

Authors and Affiliations

  1. 1.The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) and Key Laboratory of Oral Biomedicine, Ministry of Education, School and Hospital of StomatologyWuhan UniversityWuhanChina
  2. 2.Department of Oral Maxillofacial-Head Neck Oncology, School and Hospital of StomatologyWuhan UniversityWuhanChina

Personalised recommendations