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CD70 expression in tumor-associated fibroblasts predicts worse survival in colorectal cancer patients

  • Satoshi Inoue
  • Hideaki Ito
  • Takumi Tsunoda
  • Hideki Murakami
  • Masahide Ebi
  • Naotaka Ogasawara
  • Kunio Kasugai
  • Kenji Kasai
  • Hiroshi Ikeda
  • Shingo InagumaEmail author
Original Article

Abstract

The anticancer effects of immune checkpoint inhibitors against CTLA4 and CD274-PDCD1 axes are evident. However, these immunotherapies for colorectal cancers (CRCs) are now limited to a small subset of patients with microsatellite unstable tumors. Thus, therapeutics targeting other types of CRCs is desired. The CD70–CD27 axis plays a co-stimulatory role in promoting the expansion and differentiation of T-lymphocytes through the activation of NFκB pathway. Aberrant activation of the CD70–CD27 axis accelerates tumor cell proliferation, survival, and immune evasion of tumor cells. Based on these observations, drugs modulating the CD70–CD27 axis have been developed with expectation of anticancer effects. In the present study, 269 primary CRCs were evaluated immunohistochemically for CD70, CD27, and FOXP3 expression to assess their clinical usage and the application of CD70–CD27 axis modulating drugs. CRC tumor cells rarely (2.2%) expressed CD70. In contrast, tumor-surrounding fibroblasts showed various CD70 expressions (fCD70) in 14.9%. The logistic regression analysis revealed significant association of fCD70 expression with incomplete resection status (OR, 2.60; 95% CI, 1.10–6.13; P = 0.029). Overall survival was significantly decreased in the cohort of the patients with fCD70-positive tumor (P = 0.0078). Furthermore, significantly more CD27+ tumor-associated lymphocytes were detected within the primary CRCs without metastases (P = 0.024). Thus, the CD70–CD27 axis may have several roles in CRCs independent from their mismatch repair (MMR) system status. CD70–CD27 pathway-modulating therapies may be applied to CRC patients regardless of their tumor MMR status.

Keywords

Colorectal cancer (CRC) Immunohistochemistry CD70 CD27 FOXP3 

Notes

Acknowledgements

We thank Dr. Yutaka Kondo (Nagoya University) and Dr. Yoshitaka Sekido (Aichi Cancer Center Research Institute) for SW48 and ACC-MESO-1 cells, respectively. We also thank Ms. Kazuko Tanimizu, Mr. Naoki Igari, and Mr. Motoyasu Takeuchi (Aichi Medical University) for their assistance on tissue preparation and immunohistochemical staining. We had the support on the manuscript editing from Ms. Yukiko Kuru (Aichi Medical University).

Author’s contributions

Shingo Inaguma: conceived, designed, and supervised the overall study; Satoshi Inoue, Shingo Inaguma: performed molecular experiments, histological and statistical analyses, made the figures and tables, and wrote the manuscript; Hideaki Ito, Takumi Tsunoda, Hideki Murakami: performed immunohistochemical staining; Satoshi Inoue, Masahide Ebi, Naotaka Ogasawara: collected and analyzed the clinical data; Kunio Kasugai, Kenji Kasai, Hiroshi Ikeda: critically reviewed the manuscript. All authors have read and gave final approval to the submitted version.

Funding

This work was supported as a part of Grant-in-Aid for Scientific Research (C) (to Shingo Inaguma, 17K08706).

Compliance with ethical standards

This project was approved by the Institutional Ethical Review Board of Aichi Medical University Hospital.

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

428_2019_2565_MOESM1_ESM.zip (552 kb)
ESM 1 Supplementary Figure S1. Representative images for the measurement of fCD70-positive area. Photographs for fCD70 immunohistochemistry (left) were converted into black and white image (right). Black fCD70-signals were measured by ImageJ software (NIH, Bethesda, MD). This case showed 9,220 pixels, corresponding to 0.415 mm2, of fCD70-positive area. Bar; 1mm. Supplementary Figure S2. Immunoblot analyses of cultured colorectal cancer cells. All of the colon cancer cells expressed CD70 at under detectable levels. In contrast, ACC-MESO-1, a human mesothelioma cell line, expressed CD70. The following siRNAs were synthesized: siControl, 5’-GACGUAUGACUAACUAACATT-3’ and 5’-UGUUAGUUAGUCAUACGUCTT-3’; siCD70, 5’-GCAUCAGCCUGCUGCGUCUTT -3’ and 5’-AGACGCAGCAGGCUGAUGCTT-3’. After 48 hours of siRNA transfection, total lysates were prepared for immunoblot analysis. Anti-CD70 antibody was applied at a dilution of 1:1000. (ZIP 551 kb)

References

  1. 1.
    Callahan MK, Postow MA, Wolchok JD (2014) CTLA-4 and PD-1 pathway blockade: combinations in the clinic. Front Oncol 4:385Google Scholar
  2. 2.
    Luke JJ, Flaherty KT, Ribas A, Long GV (2017) Targeted agents and immunotherapies: optimizing outcomes in melanoma. Nat Rev Clin Oncol 14:463–482CrossRefGoogle Scholar
  3. 3.
    Le DT, Uram JN, Wang H et al (2015) PD-1 blockade in tumors with mismatch-repair deficiency. N Engl J Med 372:2509–2520CrossRefGoogle Scholar
  4. 4.
    Bodmer JL, Schneider P, Tschopp J (2002) The molecular architecture of the TNF superfamily. Trends Biochem Sci 27:19–26CrossRefGoogle Scholar
  5. 5.
    Locksley RM, Killeen N, Lenardo MJ (2001) The TNF and TNF receptor superfamilies: integrating mammalian biology. Cell 104:487–501CrossRefGoogle Scholar
  6. 6.
    Nolte MA, van Olffen RW, van Gisbergen KP et al (2009) Timing and tuning of CD27-CD70 interactions: the impact of signal strength in setting the balance between adaptive responses and immunopathology. Immunol Rev 229:216–231CrossRefGoogle Scholar
  7. 7.
    Duggleby RC, Shaw TN, Jarvis LB et al (2007) CD27 expression discriminates between regulatory and non-regulatory cells after expansion of human peripheral blood CD4+ CD25+ cells. Immunology 121:129–139CrossRefGoogle Scholar
  8. 8.
    Yang ZZ, Novak AJ, Ziesmer SC, Witzig TE, Ansell SM (2007) CD70+ non-Hodgkin lymphoma B cells induce Foxp3 expression and regulatory function in intratumoral CD4+CD25 T cells. Blood 110:2537–2544CrossRefGoogle Scholar
  9. 9.
    Law CL, Gordon KA, Toki BE, Yamane AK, Hering MA, Cerveny CG, Petroziello JM, Ryan MC, Smith L, Simon R, Sauter G, Oflazoglu E, Doronina SO, Meyer DL, Francisco JA, Carter P, Senter PD, Copland JA, Wood CG, Wahl AF (2006) Lymphocyte activation antigen CD70 expressed by renal cell carcinoma is a potential therapeutic target for anti-CD70 antibody-drug conjugates. Cancer Res 66:2328–2337CrossRefGoogle Scholar
  10. 10.
    Wischhusen J, Jung G, Radovanovic I, Beier C, Steinbach JP, Rimner A, Huang H, Schulz JB, Ohgaki H, Aguzzi A, Rammensee HG, Weller M (2002) Identification of CD70-mediated apoptosis of immune effector cells as a novel immune escape pathway of human glioblastoma. Cancer Res 62:2592–2599Google Scholar
  11. 11.
    Ranheim EA, Cantwell MJ, Kipps TJ (1995) Expression of CD27 and its ligand, CD70, on chronic lymphocytic leukemia B cells. Blood 85:3556–3565Google Scholar
  12. 12.
    Bertrand P, Maingonnat C, Penther D, Guney S, Ruminy P, Picquenot JM, Mareschal S, Alcantara M, Bouzelfen A, Dubois S, Figeac M, Bastard C, Tilly H, Jardin F (2013) The costimulatory molecule CD70 is regulated by distinct molecular mechanisms and is associated with overall survival in diffuse large B-cell lymphoma. Genes Chromosom Cancer 52:764–774CrossRefGoogle Scholar
  13. 13.
    Aggarwal S, He T, Fitzhugh W et al (2009) Immune modulator CD70 as a potential cisplatin resistance predictive marker in ovarian cancer. Gynecol Oncol 115:430–437CrossRefGoogle Scholar
  14. 14.
    Tanaka A, Sakaguchi S (2017) Regulatory T cells in cancer immunotherapy. Cell Res 27:109–118CrossRefGoogle Scholar
  15. 15.
    Chahlavi A, Rayman P, Richmond AL, Biswas K, Zhang R, Vogelbaum M, Tannenbaum C, Barnett G, Finke JH (2005) Glioblastomas induce T-lymphocyte death by two distinct pathways involving gangliosides and CD70. Cancer Res 65:5428–5438CrossRefGoogle Scholar
  16. 16.
    Claus C, Riether C, Schurch C, Matter MS, Hilmenyuk T, Ochsenbein AF (2012) CD27 signaling increases the frequency of regulatory T cells and promotes tumor growth. Cancer Res 72:3664–3676CrossRefGoogle Scholar
  17. 17.
    Diegmann J, Junker K, Loncarevic IF, Michel S, Schimmel B, von Eagelinq F (2006) Immune escape for renal cell carcinoma: CD70 mediates apoptosis in lymphocytes. Neoplasia 8:933–938CrossRefGoogle Scholar
  18. 18.
    Jak M, Mous R, Remmerswaal EB et al (2009) Enhanced formation and survival of CD4+ CD25hi Foxp3+ T-cells in chronic lymphocytic leukemia. Leuk Lymphoma 50:788–801CrossRefGoogle Scholar
  19. 19.
    Prasad KV, Ao Z, Yoon Y et al (1997) CD27, a member of the tumor necrosis factor receptor family, induces apoptosis and binds to Siva, a proapoptotic protein. Proc Natl Acad Sci U S A 94:6346–6351CrossRefGoogle Scholar
  20. 20.
    Yang ZZ, Grote DM, Xiu B, Ziesmer SC, Price-Troska TL, Hodge LS, Yates DM, Novak AJ, Ansell SM (2014) TGF-beta upregulates CD70 expression and induces exhaustion of effector memory T cells in B-cell non-Hodgkin's lymphoma. Leukemia 28:1872–1884CrossRefGoogle Scholar
  21. 21.
    Inaguma S, Lasota J, Felisiak-Golabek A, Kowalik A, Wang Z, Zieba S, Kalisz J, Ikeda H, Miettinen M (2017) Histopathological and genotypic characterization of metastatic colorectal carcinoma with PD-L1 (CD274)-expression: possible roles of tumour micro environmental factors for CD274 expression. J Pathol Clin Res 3:268–278CrossRefGoogle Scholar
  22. 22.
    Inaguma S, Riku M, Hashimoto M, Murakami H, Saga S, Ikeda H, Kasai K (2013) GLI1 interferes with the DNA mismatch repair system in pancreatic cancer through BHLHE41-mediated suppression of MLH1. Cancer Res 73:7313–7323CrossRefGoogle Scholar
  23. 23.
    Kanda Y (2013) Investigation of the freely available easy-to-use software 'EZR' for medical statistics. Bone Marrow Transplant 48:452–458CrossRefGoogle Scholar
  24. 24.
    Jacobs J, Deschoolmeester V, Zwaenepoel K, Flieswasser T, Deben C, van den Bossche J, Hermans C, Rolfo C, Peeters M, de Wever O, Lardon F, Siozopoulou V, Smits E, Pauwels P (2018) Unveiling a CD70-positive subset of cancer-associated fibroblasts marked by pro-migratory activity and thriving regulatory T cell accumulation. Oncoimmunology 7:e1440167CrossRefGoogle Scholar
  25. 25.
    Kalluri R (2016) The biology and function of fibroblasts in cancer. Nat Rev Cancer 16:582–598CrossRefGoogle Scholar
  26. 26.
    Mukaida N, Sasaki S (2016) Fibroblasts, an inconspicuous but essential player in colon cancer development and progression. World J Gastroenterol 22:5301–5316CrossRefGoogle Scholar
  27. 27.
    Silence K, Dreier T, Moshir M, Ulrichts P, Gabriels SME, Saunders M, Wajant H, Brouckaert P, Huyghe L, van Hauwermeiren T, Thibault A, de Haard HJ (2014) ARGX-110, a highly potent antibody targeting CD70, eliminates tumors via both enhanced ADCC and immune checkpoint blockade. MAbs 6:523–532CrossRefGoogle Scholar
  28. 28.
    Tannir NM, Forero-Torres A, Ramchandren R, Pal SK, Ansell SM, Infante JR, de Vos S, Hamlin PA, Kim SK, Whiting NC, Gartner EM, Zhao B, Thompson JA (2014) Phase I dose-escalation study of SGN-75 in patients with CD70-positive relapsed/refractory non-Hodgkin lymphoma or metastatic renal cell carcinoma. Investig New Drugs 32:1246–1257CrossRefGoogle Scholar
  29. 29.
    He LZ, Prostak N, Thomas LJ, Vitale L, Weidlick J, Crocker A, Pilsmaker CD, Round SM, Tutt A, Glennie MJ, Marsh H, Keler T (2013) Agonist anti-human CD27 monoclonal antibody induces T cell activation and tumor immunity in human CD27-transgenic mice. J Immunol 191:4174–4183CrossRefGoogle Scholar
  30. 30.
    Wasiuk A, Testa J, Weidlick J, Sisson C, Vitale L, Widger J, Crocker A, Thomas LJ, Goldstein J, Marsh HC, Keler T, He LZ (2017) CD27-mediated regulatory T cell depletion and effector T cell costimulation both contribute to antitumor efficacy. J Immunol 199:4110–4123CrossRefGoogle Scholar
  31. 31.
    Hori S, Nomura T, Sakaguchi S (2003) Control of regulatory T cell development by the transcription factor Foxp3. Science 299:1057–1061CrossRefGoogle Scholar
  32. 32.
    Ladoire S, Martin F, Ghiringhelli F (2011) Prognostic role of FOXP3+ regulatory T cells infiltrating human carcinomas: the paradox of colorectal cancer. Cancer Immunol Immunother 60:909–918CrossRefGoogle Scholar
  33. 33.
    Shang B, Liu Y, Jiang SJ (2015) Prognostic value of tumor-infiltrating FoxP3+ regulatory T cells in cancers: a systematic review and meta-analysis. Sci Rep 5:15179CrossRefGoogle Scholar
  34. 34.
    Nosho K, Baba Y, Tanaka N, Shima K, Hayashi M, Meyerhardt JA, Giovannucci E, Dranoff G, Fuchs CS, Ogino S (2010) Tumour-infiltrating T-cell subsets, molecular changes in colorectal cancer, and prognosis: cohort study and literature review. J Pathol 222:350–366CrossRefGoogle Scholar
  35. 35.
    Inaguma S, Lasota J, Wang Z, Felisiak-Golabek A, Ikeda H, Miettinen M (2017) Clinicopathologic profile, immunophenotype, and genotype of CD274 (PD-L1)-positive colorectal carcinomas. Mod Pathol 30:278–285CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Division of Gastroenterology, Department of Internal MedicineAichi Medical University School of MedicineNagakuteJapan
  2. 2.Department of PathologyAichi Medical University School of MedicineNagakuteJapan

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