Skip to main content

Advertisement

SpringerLink
Log in

Immune checkpoint CD155 promoter methylation profiling reveals cancer-associated behaviors within breast neoplasia

  • Original Article
  • Published:
Cancer Immunology, Immunotherapy Aims and scope Submit manuscript

Abstract

BACKGROUND

CD155 immune checkpoint has recently emerged as a compelling immunotherapeutic target. Epigenetic DNA methylation changes are recognized as key molecular mechanisms in cancer development. Hence, the identification of methylation markers that are sensitive and specific for breast cancer may improve early detection and predict prognosis. We speculate that CD155 promoter methylation can be a valuable epigenetic biomarker, based upon strong indications for its immunoregulatory functions.

METHODS

Methylation analyses were conducted on 14 CpGs sites in the CD155 promoter region by bisulfite pyrosequencing. To elucidate the related gene expression changes, a transcriptional study using RT-qPCR was performed. Statistical analyses were performed to evaluate correlations of CD155 methylation profiles with mRNA expression together with clinical-pathological features, prognosis and immune infiltrate.

RESULTS

CD155 promoter methylation profile was significantly associated with SBR grade, tumor size, molecular subgroups, HER2 and hormonal receptors expression status. Low CD155 methylation rates correlated with better prognosis in univariate cox proportional hazard analysis and appeared as an independent survival predictor in cox-regression multivariate analysis. Further, methylation changes at CD155 specific CpG sites were consistent with CD155 membranous mRNA isoform expression status. Statistical analyses also showed a significant association with immune Natural Killer cell infiltrate when looking at the CpG7, CpG8, CpG9 and CpG11 sites.

CONCLUSION

Altogether, our results contribute to a better understanding of the impact of CD155 immune checkpoint modality expression in breast tumors, revealing for the first time that specific CpG sites from CD155 promoter may be a potential biomarker in breast cancer monitoring.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Availability of data and material

All data generated or analyzed during this study are included in this published article.

Abbreviations

ACTB:

Beta-Actin

bp:

Base pair

cyt-CD155:

Cytoplasmic CD155

DFS:

Disease-Free Survival

FFPE:

Formalin-Fixed and Paraffin-Embedded

HER2:

HER2 positive

kbp:

Kilobase pair

LA:

Luminal A

LB-Like:

Luminal B like

m-CD155:

Membranous CD155

NK:

Natural Killer

NK-TILs:

Tumor-Infiltrating Natural Killer Cell

OS:

Overall Survival

RT-qPCR:

Real-time Quantitative Reverse Transcriptase Polymerase Chain Reaction

SBR:

Scarff-Bloom-Richardson

TBE:

Tris-Borate-EDTA

TILs:

Tumor-Infiltrating Lymphocytes

TME:

Tumor Microenvironment

TNBC:

Triple Negative breast cancer

TNM:

Tumor lymph node and metastases

References

  1. Pardoll DM (2012) The blockade of immune checkpoints in cancer immunotherapy. Nat Rev Cancer 12(4):252–264. https://doi.org/10.1038/nrc3239

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Topalian SL, Drake CG, Pardoll DM (2015) Immune checkpoint blockade: a common denominator approach to cancer therapy. Cancer Cell 27(4):450–461. https://doi.org/10.1016/j.ccell.2015.03.001

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Sharma P, Allison JP (2015) The future of immune checkpoint therapy. Science 348(6230):56–61. https://doi.org/10.1126/science.aaa8172

    Article  CAS  PubMed  Google Scholar 

  4. Chaudhary B, Elkord E (2016) Regulatory T Cells in the Tumor Microenvironment and Cancer Progression: Role and Therapeutic Targeting. Vaccines (Basel). https://doi.org/10.3390/vaccines4030028

    Article  Google Scholar 

  5. Sasidharan Nair V, Elkord E (2018) Immune checkpoint inhibitors in cancer therapy: a focus on T-regulatory cells. Immunol Cell Biol 96(1):21–33. https://doi.org/10.1111/imcb.1003

    Article  CAS  PubMed  Google Scholar 

  6. Dougall WC, Kurtulus S, Smyth MJ, Anderson AC (2017) TIGIT and CD96: new checkpoint receptor targets for cancer immunotherapy. Immunol Rev 276(1):112–120. https://doi.org/10.1111/imr.12518

    Article  CAS  PubMed  Google Scholar 

  7. Chan CJ et al (2014) The receptors CD96 and CD226 oppose each other in the regulation of natural killer cell functions. Nat Immunol 15(5):431–438. https://doi.org/10.1038/ni.2850

    Article  CAS  PubMed  Google Scholar 

  8. Masson D et al (2001) Overexpression of the CD155 gene in human colorectal carcinoma. Gut 49(2):236–240

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Nakai R et al (2010) Overexpression of Necl-5 correlates with unfavorable prognosis in patients with lung adenocarcinoma. Cancer Sci 101(5):1326–1330. https://doi.org/10.1111/j.1349-7006.2010.01530.x

    Article  CAS  PubMed  Google Scholar 

  10. Bevelacqua V et al (2012) Nectin like-5 overexpression correlates with the malignant phenotype in cutaneous melanoma. Oncotarget 3(8):882–892. https://doi.org/10.18632/oncotarget.594

    Article  PubMed  PubMed Central  Google Scholar 

  11. Nishiwada S et al (2015) Clinical significance of CD155 expression in human pancreatic cancer. Anticancer Res 35(4):2287–2297

    CAS  PubMed  Google Scholar 

  12. Sloan KE, Stewart JK, Treloar AF, Matthews RT, Jay DG (2005) CD155/PVR enhances glioma cell dispersal by regulating adhesion signaling and focal adhesion dynamics. Cancer Res 65(23):10930–10937. https://doi.org/10.1158/0008-5472.CAN-05-1890

    Article  CAS  PubMed  Google Scholar 

  13. Stamm H et al (2019) Targeting the TIGIT-PVR immune checkpoint axis as novel therapeutic option in breast cancer. Oncoimmunology 8(12):e1674605. https://doi.org/10.1080/2162402X.2019.1674605

    Article  PubMed  PubMed Central  Google Scholar 

  14. Triki H et al (2019) CD155 expression in human breast cancer: Clinical significance and relevance to natural killer cell infiltration. Life Sci 231:116543. https://doi.org/10.1016/j.lfs.2019.116543

    Article  CAS  PubMed  Google Scholar 

  15. Y.-C. Li et al., (2020) Overexpression of an Immune Checkpoint (CD155) in Breast Cancer Associated with Prognostic Significance and Exhausted Tumor-Infiltrating Lymphocytes: A Cohort Study. Journal of Immunology Research. https://www.hindawi.com/journals/jir/2020/3948928/. Accessed 08 Apr 2020

  16. Excoffon KJDA, Bowers JR, Sharma P (2014) 1. Alternative splicing of viral receptors: A review of the diverse morphologies and physiologies of adenoviral receptors. Recent Res Dev Virol 9:1–24

    PubMed  PubMed Central  Google Scholar 

  17. Koike S et al (1990) The poliovirus receptor protein is produced both as membrane-bound and secreted forms. EMBO J 9(10):3217–3224

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Béatrice B et al (2003) Identification of secreted CD155 isoforms. BiochemBiophys Res Commun. 309(1):175–182. https://doi.org/10.1016/s0006-291x(03)01560-2

    Article  Google Scholar 

  19. Lozano E, Dominguez-Villar M, Kuchroo V, Hafler DA (2012) The TIGIT/CD226 axis regulates human T cell function. J Immunol 188(8):3869–3875. https://doi.org/10.4049/jimmunol.1103627

    Article  CAS  PubMed  Google Scholar 

  20. Kulis M, Esteller M (2010) DNA methylation and cancer. Adv Genet 70:27–56. https://doi.org/10.1016/B978-0-12-380866-0.60002-2

    Article  PubMed  Google Scholar 

  21. Sinčić N, Herceg Z (2011) DNA methylation and cancer: ghosts and angels above the genes. CurrOpin Oncol 23(1):69–76. https://doi.org/10.1097/CCO.0b013e3283412eb4

    Article  CAS  Google Scholar 

  22. Wu Y, Sarkissyan M, Vadgama JV (2015) Epigenetics in breast and prostate cancer. Methods Mol Biol 1238:425–466. https://doi.org/10.1007/978-1-4939-1804-1_23

    Article  PubMed  PubMed Central  Google Scholar 

  23. Sasidharan Nair V, El Salhat H, Taha RZ, John A, Ali BR, Elkord E (2018) DNA methylation and repressive H3K9 and H3K27 trimethylation in the promoter regions of PD-1, CTLA-4, TIM-3, LAG-3, TIGIT, and PD-L1 genes in human primary breast cancer. Clin Epigenetics. https://doi.org/10.1186/s13148-018-0512-1

    Article  PubMed  PubMed Central  Google Scholar 

  24. Roulois D, Yau HL, De Carvalho DD (2016) Pharmacological DNA demethylation: Implications for cancer immunotherapy. Oncoimmunology 5(3):e1090077. https://doi.org/10.1080/2162402X.2015.1090077

    Article  CAS  PubMed  Google Scholar 

  25. Maruvada P, Wang W, Wagner PD, Srivastava S (2005) Biomarkers in molecular medicine: cancer detection and diagnosis. Biotechniques. https://doi.org/10.2144/05384su04

    Article  PubMed  Google Scholar 

  26. Wolff AC et al (2013) Recommendations for human epidermal growth factor receptor 2 testing in breast cancer: American Society of Clinical Oncology/College of American Pathologists clinical practice guideline update. J Clin Oncol 31(31):3997–4013. https://doi.org/10.1200/JCO.2013.50.9984

    Article  PubMed  Google Scholar 

  27. Bouzidi L et al (2021) Prognostic value of natural killer cells besides tumor infiltrating lymphocytes in breast cancer tissues. Clin Breast Cancer. https://doi.org/10.1016/j.clbc.2021.02.003

    Article  PubMed  Google Scholar 

  28. Sambrook J, Russell DW (2006) Purification of nucleic acids by extraction with phenol:chloroform. CSH Protoc. 2006(1):169–170. https://doi.org/10.1101/pdb.prot4455

    Article  Google Scholar 

  29. Saiki RK et al (1988) Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239(4839):487–491

    Article  CAS  PubMed  Google Scholar 

  30. Declerck K et al (2017) Interaction between prenatal pesticide exposure and a common polymorphism in the PON1 gene on DNA methylation in genes associated with cardio-metabolic disease risk—an exploratory study. Clin Epigenetics. https://doi.org/10.1186/s13148-017-0336-4

    Article  PubMed  PubMed Central  Google Scholar 

  31. Sun Y et al (2020) Combined evaluation of the expression status of CD155 and TIGIT plays an important role in the prognosis of LUAD (lung adenocarcinoma). Int Immunopharmacol 80:106198. https://doi.org/10.1016/j.intimp.2020.106198

    Article  CAS  PubMed  Google Scholar 

  32. Jones PA, Baylin SB (2007) The epigenomics of cancer. Cell 128(4):683–692. https://doi.org/10.1016/j.cell.2007.01.029

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Jones PA (2012) Functions of DNA methylation: islands, start sites, gene bodies and beyond. Nat Rev Genet 13(7):484–492. https://doi.org/10.1038/nrg3230

    Article  CAS  PubMed  Google Scholar 

  34. Gibney ER, Nolan CM (2010) Epigenetics and gene expression. Heredity. https://doi.org/10.1038/hdy.2010.54

    Article  PubMed  Google Scholar 

  35. Chen G et al (2002) Discordant protein and mRNA expression in lung adenocarcinomas. Mol Cell Proteomics 1(4):304–313. https://doi.org/10.1074/mcp.M200008-MCP200

    Article  CAS  PubMed  Google Scholar 

  36. Yong H et al (2019) CD155 expression and its prognostic value in postoperative patients with breast cancer. Biomed Pharmacother 115:108884. https://doi.org/10.1016/j.biopha.2019.108884

    Article  CAS  PubMed  Google Scholar 

  37. Atsumi S, Matsumine A, Toyoda H, Niimi R, Iino T, Sudo A (2013) Prognostic significance of CD155 mRNA expression in soft tissue sarcomas. Oncol Lett 5(6):1771–1776. https://doi.org/10.3892/ol.2013.1280

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Zhao K, Ma L, Feng L, Huang Z, Meng X, Yu J (2021) CD155 Overexpression Correlates With Poor Prognosis in Primary Small Cell Carcinoma of the Esophagus. Front Mol Biosci. https://doi.org/10.3389/fmolb.2020.608404

    Article  PubMed  PubMed Central  Google Scholar 

  39. Wang R-B et al (2020) Overexpression of CD155 is associated with PD-1 and PD-L1 expression on immune cells, rather than tumor cells in the breast cancer microenvironment. World J Clin Cases 8(23):5935–5943. https://doi.org/10.12998/wjcc.v8.i23.5935

    Article  PubMed  PubMed Central  Google Scholar 

  40. “The Cancer Genome Atlas Program - National Cancer Institute,” Jun. 13, 2018. https://www.cancer.gov/about-nci/organization/ccg/research/structural-genomics/tcga. Accessed 20 Feb 2021

  41. Koch A, De Meyer T, Jeschke J, Van Criekinge W (2015) MEXPRESS: visualizing expression, DNA methylation and clinical TCGA data. BMC Genomics 16:636. https://doi.org/10.1186/s12864-015-1847-z

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. O’Donnell JS, Long GV, Scolyer RA, Teng MWL, Smyth MJ (2017) Resistance to PD1/PDL1 checkpoint inhibition. Cancer Treat Rev 52:71–81. https://doi.org/10.1016/j.ctrv.2016.11.007

    Article  CAS  PubMed  Google Scholar 

  43. O’Donnell JS, Madore J, Li X-Y, Smyth MJ (2020) Tumor intrinsic and extrinsic immune functions of CD155. Semin Cancer Biol 65:189–196. https://doi.org/10.1016/j.semcancer.2019.11.013

    Article  CAS  PubMed  Google Scholar 

  44. Lupo KB, Matosevic S (2020) CD155 immunoregulation as a target for natural killer cell immunotherapy in glioblastoma. J Hematol Oncol 13(1):76. https://doi.org/10.1186/s13045-020-00913-2

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Chauvin J-M et al (2015) TIGIT and PD-1 impair tumor antigen–specific CD8+ T cells in melanoma patients. J Clin Invest 125(5):2046–2058. https://doi.org/10.1172/JCI80445

    Article  PubMed  PubMed Central  Google Scholar 

  46. Smazynski J et al (2020) The immune suppressive factors CD155 and PD-L1 show contrasting expression patterns and immune correlates in ovarian and other cancers. Gynecol Oncol 158(1):167–177. https://doi.org/10.1016/j.ygyno.2020.04.689

    Article  CAS  PubMed  Google Scholar 

  47. Huang D-W, Huang M, Lin X-S, Huang Q (2017) CD155 expression and its correlation with clinicopathologic characteristics, angiogenesis, and prognosis in human cholangiocarcinoma. Onco Targets Ther 10:3817–3825. https://doi.org/10.2147/OTT.S141476

    Article  PubMed  PubMed Central  Google Scholar 

  48. Qu P et al (2015) Loss of CD155 expression predicts poor prognosis in hepatocellular carcinoma. Histopathology 66(5):706–714. https://doi.org/10.1111/his.12584

    Article  PubMed  Google Scholar 

Download references

Acknowledgements

We thank our study participants for their contribution to this study. A further thanks goes to the Protein Chemistry, Proteomics and Epigenetic Signaling (PPES), University of Antwerp, team members for their collaboration and valuable contribution. This work was partially supported by ISESCO (Islamic Educational, Scientific and Cultural Organization) Research grant (Ref No. 2148).

Funding

This work was financially supported by ISESCO (Islamic Educational, Scientific and Cultural Organization) Research grant (Ref N°2148).

Author information

Authors and Affiliations

  1. Laboratory of Molecular and Cellular Screening Processes, Centre de Biotechnologie de Sfax, University of Sfax, B.P 1177, 3018, Sfax, Tunisia

    Hana Triki, Ahmed Rebai & Boutheina Cherif

  2. Laboratory of Protein Chemistry, Proteomics and Epigenetic Signaling (PPES) and Integrated Personalizedand Precision Oncology Network (IPPON), Department of Biomedical Sciences, University of Antwerp, Antwerp, Belgium

    Ken Declerck & Wim Vanden Berghe

  3. Department of Pathology, University Hospital Habib Bourguiba, Sfax, Tunisia

    Slim Charfi & Tahya Sellami

  4. Department of Medical Oncology, University Hospital Habib Bourguiba, Sfax, Tunisia

    Wala Ben Kridis

  5. Department of Gynecology, University Hospital Hédi Chaker, Sfax, Tunisia

    Kais Chaabane & Sawssan Ben Halima

Authors
  1. Hana Triki
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ken Declerck
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Slim Charfi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Wala Ben Kridis
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Kais Chaabane
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Sawssan Ben Halima
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Tahya Sellami
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Ahmed Rebai
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Wim Vanden Berghe
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Boutheina Cherif
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

HT contributed to data curation, formal analysis, investigation, methodology, software, validation, visualization, writing—original draft, writing—review and editing. KD contributed to formal analysis, software, visualization. SC contributed to data curation, project administration, resources, supervision, validation. KC contributed to data curation, resources. SBH contributed to data curation, resources. WBK contributed to data curation, resources. TS contributed to data curation, resources. AR contributed to formal analysis, project administration, supervision, validation. WVB contributed to project administration, resources, supervision, validation, visualization, writing—review and editing. BC contributed to conceptualization, funding acquisition, investigation, methodology, project administration, resources, supervision, validation, visualization, writing—review and editing.

Corresponding author

Correspondence to Boutheina Cherif.

Ethics declarations

Conflicts of interest

The authors have no conflicts of interest to declare.

Ethics approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and the national research committee of Habib Bourguiba University Hospital and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. Sampling was made only on patient tissues from tissue library of Pathology Department-Habib Bourguiba Hospital, and no samples were made specifically for the study.

Consent to participate

We have conducted a retrospective study, for this type of study, formal consent is not required.

Consent for publication

All authors reviewed and approved the manuscript for submission.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Triki, H., Declerck, K., Charfi, S. et al. Immune checkpoint CD155 promoter methylation profiling reveals cancer-associated behaviors within breast neoplasia. Cancer Immunol Immunother 71, 1139–1155 (2022). https://doi.org/10.1007/s00262-021-03064-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00262-021-03064-6

Keywords

Search

Navigation