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Clinical relevance of the comparative expression of immune checkpoint markers with the clinicopathological findings in patients with primary and chemoreduced retinoblastoma

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Abstract

Purpose

The goal of this study is to identify the pathological findings and expression of immune checkpoint marker (PD-1, PD-L1, and CTLA-4) in the tumor microenvironment of both primary and chemoreduced retinoblastoma and correlate them with clinicopathological parameters and patient outcome.

Methods

Total of 262 prospective cases was included prospectively in which 144 cases underwent primary enucleation and 118 cases received chemotherapy/radiotherapy before enucleation (chemoreduced retinoblastoma). Immunohistochemistry, qRT-PCR and western blotting were performed to evaluate the expression pattern of immune checkpoint markers in primary and chemoreduced retinoblastoma.

Results

Tumor microenvironment were different for both primary and chemoreduced retinoblastoma. Expression of PD-1 was found in 29/144 (20.13%) and 48/118 (40.67%) in primary and chemoreduced retinoblastoma, respectively, whereas PD-L1 was expressed in 46/144 (31.94%) and 22/118 (18.64%) in cases of primary and chemoreduced retinoblastoma, respectively. Expression pattern of CTLA-4 protein was similar in both groups of retinoblastoma. On multivariate analysis, massive choroidal invasion, bilaterality and PD-L1 expression (p = 0.034) were found to be statistically significant factors in primary retinoblastoma, whereas PD-1 expression (p = 0.015) and foamy macrophages were significant factors in chemoreduced retinoblastoma. Overall survival was reduced in cases of PD-L1 (80.76%) expressed primary retinoblastoma, and PD-1 (63.28%) expressed chemoreduced retinoblastoma.

Conclusions

This is the first of its kind study predicting a relevant role of the immune checkpoint markers in both groups of primary and chemoreduced retinoblastoma with prognostic significance. Differential expression of these markers in both group of retinoblastoma is a novel finding and might be an interesting and beneficial target for chemoresistant tumors.

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Abbreviations

AJCC:

American Joint Committee on Cancer

CTLA-4:

Cytotoxic T-lymphocyte-associated antigen-4

DAB:

3,3′-Diaminobenzidine

IHC:

Immunohistochemistry

PD-1:

Programmed death-1

PD-L1:

Programmed death-ligand 1

qRT-PCR:

Quantitative real-time polymerase chain reaction

Rb:

Retinoblastoma

SDS–PAGE:

Sodium dodecyl sulfate–polyacrylamide gel electrophoresis

TILs:

Tumor-infiltrating lymphocytes

References

  1. Singh L, Kashyap S (2018) Update on pathology of retinoblastoma. Int J Ophthalmol 11(12):2011–2016

    PubMed  PubMed Central  Google Scholar 

  2. Chawla B, Singh R (2017) Recent advances and challenges in the management of retinoblastoma. Indian J Ophthalmol 65(2):133–139

    PubMed  PubMed Central  Google Scholar 

  3. Ghassemi F, Khodabande A (2015) Risk definition and management strategies in retinoblastoma: current perspectives. Clin ophthalmol 9:985–994

    CAS  PubMed  PubMed Central  Google Scholar 

  4. Yanık Ö, Gündüz K, Yavuz K et al (2015) Chemotherapy in retinoblastoma: current approaches. Turk J Ophthalmol 45(6):259–267

    PubMed  PubMed Central  Google Scholar 

  5. Gombos DS, Hungerford J, Abramson DH et al (2007) Secondary acute myelogenous leukemia in patients with retinoblastoma: is chemotherapy a factor? Ophthalmology 114(7):1378–1383

    PubMed  Google Scholar 

  6. Quail DF, Joyce JA (2013) Microenvironmental regulation of tumor progression and metastasis. Nat med 19(11):1423–1437

    CAS  PubMed  PubMed Central  Google Scholar 

  7. Chen F, Zhuang X, Lin L et al (2015) New horizons in tumor microenvironment biology: challenges and opportunities. BMC Med 13(1):45

    PubMed  PubMed Central  Google Scholar 

  8. Wang M, Zhao J, Zhang L et al (2017) Role of tumor microenvironment in tumorigenesis. J Cancer 8(5):761–773

    CAS  PubMed  PubMed Central  Google Scholar 

  9. Sachdeva UM, O’Brien JM (2012) Understanding pRb: toward the necessary development of targeted treatments for retinoblastoma. J Clin Invest 122(2):425–434

    CAS  PubMed  PubMed Central  Google Scholar 

  10. Fang H, Declerck YA (2013) Targeting the tumor microenvironment: from understanding pathways to effective clinical trials. Cancer Res 73(16):4695–4777

    Google Scholar 

  11. Shih K, Arkenau HT, Infante JR (2014) Clinical impact of checkpoint inhibitors as novel cancer therapies. Drugs 74(17):1993–2013

    CAS  PubMed  PubMed Central  Google Scholar 

  12. Seidel JA, Otsuka A, Kabashima K (2018) Anti-PD-1 and Anti-CTLA-4 Therapies in cancer: mechanisms of action, efficacy, and limitations. Front Oncol 8:86

    PubMed  PubMed Central  Google Scholar 

  13. Hanahan D, Weinberg RA (2011) Hallmarks of cancer: the next generation. Cell 144(5):646–674

    CAS  PubMed  Google Scholar 

  14. Henick BS, Herbst RS, Goldberg SB (2014) The PD-1 pathway as a therapeutic target to overcome immune escape mechanisms in cancer. Expert Opin Ther Targets 18(12):1407–1420

    CAS  PubMed  Google Scholar 

  15. Yang W, Li H, Chen PW et al (2009) PD-L1 expression on human ocular cells and its possible role in regulating immune-mediated ocular inflammation. Invest Ophthalmol Vis Sci 50(1):273–280

    PubMed  Google Scholar 

  16. Alsaab HO, Sau S, Alzhrani R et al (2017) PD-1 and PD-L1 checkpoint signaling inhibition for cancer immunotherapy: mechanism, combinations, and clinical outcome. Front Pharmacol 8:561

    PubMed  PubMed Central  Google Scholar 

  17. Udall M, Rizzo M, Kenny J et al (2018) PD-L1 diagnostic tests: a systematic literature review of scoring algorithms and test-validation metrics. Diagn Pathol 13(1):12

    PubMed  PubMed Central  Google Scholar 

  18. Juneja VR, McGuire KA, Manguso RT et al (2017) PD-L1 on tumor cells is sufficient for immune evasion in immunogenic tumors and inhibits CD8 T cell cytotoxicity. J Exp Med 214(4):895–904

    CAS  PubMed  PubMed Central  Google Scholar 

  19. Wang Q, Liu F, Liu L (2017) Prognostic significance of PD-L1 in solid tumor: an updated meta-analysis. Medicine 96(18):e6369

    CAS  PubMed  PubMed Central  Google Scholar 

  20. Pianko MJ, Liu Y, Bagchi S et al (2017) Immune checkpoint blockade for hematologic malignancies: a review. Stem Cell Investig 4:32

    PubMed  PubMed Central  Google Scholar 

  21. Mallipatna A, Gallie BL, Chévez-Barrios P (2017) Retinoblastoma. In: Amin MB, Edge SB, Greene FL (eds) AJCC Cancer Staging Manual, 8th edn. Springer, New York, NY, pp 819–831

    Google Scholar 

  22. Di Nicolantonio F, Neale M, Onadim Z et al (2003) The chemosensitivity profile of retinoblastoma. In: Chemosensitivity testing in oncology. Springer, Berlin, pp 73–80

  23. Radhakrishnan V, Kashyap S, Pushker N et al (2012) Outcome, pathologic findings, and compliance in orbital retinoblastoma (international retinoblastoma staging system stage III) treated with neoadjuvant chemotherapy: a prospective study. Ophthalmology 119(7):1470–1477

    PubMed  Google Scholar 

  24. Usui Y, Okunuki Y, Hattori T et al (2006) Expression of costimulatory molecules on human retinoblastoma cells Y-79: functional expression of CD40 and B7H1. Invest Ophthalmol Vis Sci 47(10):4607–4613

    PubMed  Google Scholar 

  25. Li H, Li X, Liu S et al (2017) Programmed cell death-1 (PD-1) checkpoint blockade in combination with a mammalian target of rapamycin inhibitor restrains hepatocellular carcinoma growth induced by hepatoma cell-intrinsic PD-1. Hepatology 66(6):1920–1933

    CAS  PubMed  Google Scholar 

  26. Kleffel S, Posch C, Barthel SR et al (2015) Melanoma cell-intrinsic PD-1 receptor functions promote tumor growth. Cell 162(6):1242–1256

    CAS  PubMed  PubMed Central  Google Scholar 

  27. Du S, McCall N, Park K et al (2018) Blockade of tumor-expressed PD-1 promotes lung cancer growth. Oncoimmunology 7(4):e1408747

    PubMed  PubMed Central  Google Scholar 

  28. Yao H, Wang H, Li C et al (2018) Cancer cell-intrinsic PD-1 and implications in combinatorial immunotherapy. Front Immunol 9:1774

    PubMed  PubMed Central  Google Scholar 

  29. Raguraman R, Parameswaran S, Kanwar JR et al (2019) Evidence of tumour microenvironment and stromal cellular components in retinoblastoma. Ocul Oncol Pathol 5(2):85–93

    PubMed  Google Scholar 

  30. Lotfi R, Kaltenmeier C, Lotze MT et al (2016) Until death do us part: necrosis and oxidation promote the tumor microenvironment. Transfus Med Hemother 43(2):120–132

    PubMed  PubMed Central  Google Scholar 

  31. Chong EM, Coffee RE, Chintagumpala M et al (2006) Extensively necrotic retinoblastoma is associated with high-risk prognostic factors. Arch Pathol Lab Med 130(11):1669–1672

    PubMed  Google Scholar 

  32. Lotfi R, Schrezenmeier H, Lotze MT (2009) Immunotherapy for cancer: promoting innate immunity. Front Biosci 14:818–832

    CAS  Google Scholar 

  33. Thompson RH, Kuntz SM, Leibovich BC et al (2006) Tumor B7–H1 is associated with poor prognosis in renal cell carcinoma patients with long-term follow-up. Cancer Res 66(7):3381–3385

    CAS  PubMed  Google Scholar 

  34. Gajewski TF, Schreiber H, Fu YX (2013) Innate and adaptive immune cells in the tumor microenvironment. Nat immunol 14(10):1014–1022

    CAS  PubMed  PubMed Central  Google Scholar 

  35. Richards DM, Hettinger J, Feuerer M (2013) Monocytes and macrophages in cancer: development and functions. Cancer Microenviron 6(2):179–191

    CAS  PubMed  Google Scholar 

  36. de Palma M, Lewis CE (2013) Macrophage regulation of tumor responses to anticancer therapies. Cancer Cell 23(3):277–286

    PubMed  Google Scholar 

  37. Demirci H, Eagle RC, Shields CL et al (2003) Histopathologic findings in eyes with retinoblastoma treated only with chemoreduction. Arch Ophthalmol 121(8):1125–1131

    PubMed  Google Scholar 

  38. Pollard JW (2008) Macrophages define the invasive microenvironment in breast cancer. J Leukoc Biol 84(3):623–630

    CAS  PubMed  PubMed Central  Google Scholar 

  39. Long M, Beckwith K, Do P et al (2017) Ibrutinib treatment improves T cell number and function in CLL patients. J Clin Invest 127(8):3052–3064

    PubMed  PubMed Central  Google Scholar 

  40. Mesnage SJ, Auguste A, Genestie C et al (2016) Neoadjuvant chemotherapy (NACT) increases immune infiltration and programmed death-ligand 1 (PD-L1) expression in epithelial ovarian cancer (EOC). Ann of Oncol 28(3):651–657

    Google Scholar 

  41. Katsuya Y, Horinouchi H, Asao T et al (2016) Expression of programmed death 1 (PD-1) and its ligand (PD-L1) in thymic epithelial tumors: impact on treatment efficacy and alteration in expression after chemotherapy. Lung Cancer 99:4–10

    PubMed  Google Scholar 

  42. Kim HS, Kim JY, Lee YJ et al (2018) Expression of programmed cell death ligand 1 and immune checkpoint markers in residual tumors after neoadjuvant chemotherapy for advanced high-grade serous ovarian cancer. Gynecol oncol 151(3):414–421

    CAS  PubMed  Google Scholar 

  43. Sheng J, Fang W, Yu J et al (2016) Expression of programmed death ligand-1 on tumor cells varies pre and post chemotherapy in non-small cell lung cancer. Sci Rep 6:20090

    CAS  PubMed  PubMed Central  Google Scholar 

  44. Meng Y, Liang H, Hu J et al (2018) PD-L1 Expression correlates with tumor infiltrating lymphocytes and response to neoadjuvant chemotherapy in cervical cancer. J Cancer 9(16):2938–2945

    PubMed  PubMed Central  Google Scholar 

  45. Wimberly H, Brown JR, Schalper K et al (2015) PD-L1 expression correlates with tumor-infiltrating lymphocytes and response to neoadjuvant chemotherapy in breast cancer. Cancer Immunol Res 3(4):326–332

    CAS  PubMed  Google Scholar 

  46. Chae YK, Arya A, Iams W et al (2018) Current landscape and future of dual anti-CTLA4 and PD-1/PD-L1 blockade immunotherapy in cancer; lessons learned from clinical trials with melanoma and non-small cell lung cancer (NSCLC). J Immunother Cancer 6(1):39

    PubMed  PubMed Central  Google Scholar 

  47. Chen X, Kunda PE, Lin J et al (2018) SYK-targeted dendritic cell-mediated cytotoxic T lymphocytes enhance the effect of immunotherapy on retinoblastoma. J Cancer Res Clin Oncol 144(4):675–684

    CAS  PubMed  PubMed Central  Google Scholar 

  48. Singh L, Singh MK, Rizvi MA, Pushker N, Sen S, Kashyap S (2019) Differential expression patterns of immune checkpoint markers in tumour-stromal microenvironment of primary and chemoreduced retinoblastoma. Ann Oncol 30(11):447003

    Google Scholar 

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Acknowledgements

We are very grateful to Mr. Pankaj Kumar for his excellent technical assistance.

Funding

The work was supported by Department for Science and Technology (DST), Govt. of India for providing National Post-Doctoral fellowship (N-PDF) to Dr. Lata Singh and conducting this research (NPDF/2016/000903).

Author information

Author notes

  1. Lata Singh and Seema Kashyap have equally contributed.

Authors and Affiliations

  1. Department of Biosciences, Jamia Millia Islamia, New Delhi, India

    Lata Singh & Moshahid Alam Rizvi

  2. Department of Ocular Pathology, Dr. R. P. Centre for Ophthalmic Sciences, All India Institute of Medical Sciences, New Delhi, 110029, India

    Lata Singh, Mithalesh Kumar Singh, Seema Sen & Seema Kashyap

  3. Department of Medical Oncology, IRCH, All India Institute of Medical Sciences, New Delhi, India

    Sameer Bakhshi

  4. Department of Ophthalmology, Dr. R. P. Center for Ophthalmic Sciences, All India Institute of Medical Sciences, New Delhi, India

    Rachna Meel & Neiwete Lomi

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  1. Lata Singh
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Contributions

LS and MR were responsible for the conception and design of the work; MKS contributed the acquisition, analysis, and interpretation of data for the work; SB helped in the follow-up of the patients; RM and NL recruited the patients and provided the tissue samples; SS helped in reviewing the histopathology slides; All authors reviewed and approved the final version of the manuscript.

Corresponding author

Correspondence to Seema Kashyap.

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Conflict of interest

The author(s) have no proprietary or commercial interest in any materials discussed in this article.

Ethical approval

Ethical approval was obtained from Institute’s Ethical Committee, All India Institute of Medical Sciences (Ref. No. IEC-424/RP-6/2016) and carried out in accordance with the Declaration of Helsinki principles.

Informed consent

Written consent was obtained from their legal guardians of all the patients for collection of tissue samples prior to the surgery.

Participants

This research involved the human participants.

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Singh, L., Singh, M.K., Rizvi, M.A. et al. Clinical relevance of the comparative expression of immune checkpoint markers with the clinicopathological findings in patients with primary and chemoreduced retinoblastoma. Cancer Immunol Immunother 69, 1087–1099 (2020). https://doi.org/10.1007/s00262-020-02529-4

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  • DOI: https://doi.org/10.1007/s00262-020-02529-4

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