Immune cell populations in the tumour environment following calcium electropora­tion for cutaneous metastasis: a histopathological study

Authors

  • Mille Vissing Centre for Experimental Drug and Gene Electrotransfer (C*EDGE), Department of Clinical Oncology and Palliative Care, Zealand University Hospital, Roskilde and Næstved, Næstved, Denmark; Department of Clinical Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark
  • Sandra Sinius Pouplier Department of Clinical Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark; Department of Surgical Pathology, Zealand University Hospital, Roskilde, Denmark
  • Lars Munch Larsen Centre for Experimental Drug and Gene Electrotransfer (C*EDGE), Department of Clinical Oncology and Palliative Care, Zealand University Hospital, Næstved, Denmark
  • Stine Krog Frandsen Centre for Experimental Drug and Gene Electrotransfer (C*EDGE), Department of Clinical Oncology and Palliative Care, Zealand University Hospital, Næstved, Denmark
  • Alexey Lodin Centre for Experimental Drug and Gene Electrotransfer (C*EDGE), Department of Clinical Oncology and Palliative Care, Zealand University Hospital, Næstved, Denmark
  • Anne-Vibeke Lænkholm Department of Clinical Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark; Department of Surgical Pathology, Zealand University Hospital, Roskilde, Denmark
  • Julie Gehl Centre for Experimental Drug and Gene Electrotransfer (C*EDGE), Department of Clinical Oncology and Palliative Care, Zealand University Hospital, Roskilde and Næstved, Næstved, Denmark; Department of Clinical Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark; bDepartment of Clinical Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark

DOI:

https://doi.org/10.2340/1651-226X.2024.19462

Keywords:

Calcium electroporation, Cutaneous metastases, Histopathology, Tumour microenvironment, Clinical trial

Abstract

Background and Purpose: Calcium electroporation (CaEP) involves injecting calcium into tumour tissues and using electrical pulses to create membrane pores that induce cell death. This study assesses resultant immune responses and histopathological changes in patients with cutaneous metastases.

Patients/Materials and Methods: The aimed cohort comprised 24 patients with metastases exceeding 5 mm. Tumours were treated once with CaEP (day 0) or twice (day 28). Biopsies were performed on days 0 and 2, with additional samples on days 7, 28, 30, 35, 60, and 90 if multiple tumours were treated. The primary endpoint was the change in tumour-infiltrating lymphocytes (TILs) two days post-treatment, with secondary endpoints evaluating local and systemic immune responses via histopathological analysis of immune markers, necrosis, and inflammation.

Results: Seventeen patients, with metastases primarily from breast cancer (14 patients), but also lung cancer (1), melanoma (1), and urothelial cancer (1), completed the study. Of the 49 lesions treated, no significant changes in TIL count or PD-L1 expression were observed. However, there was substantial necrosis and a decrease in FOXP3-expression (p = 0.0025) noted, with a slight increase in CD4+ cells but no changes in CD3, CD8, or CD20 expressions. Notably, four patients showed reduced tumour invasiveness, including one case of an abscopal response.

Interpretation: This exploratory study indicates that CaEP can be an effective anti-tumour therapy potentially enhancing immunity. Significant necrosis and decreased regulatory lymphocytes were observed, although TIL count remained unchanged. Several patients exhibited clinical signs of immune response following treatment.

Downloads

Download data is not yet available.

References

Frandsen SK, Gissel H, Hojman P, et al. Direct therapeutic applications of calcium electroporation to effectively induce tumor necrosis. Cancer Res. 2012;72(6):1336–41.

https://doi.org/10.1158/0008-5472.CAN-11-3782 DOI: https://doi.org/10.1158/0008-5472.CAN-11-3782

Hoejholt KL, Mužić T, Jensen SD, et al. Calcium electroporation and electrochemotherapy for cancer treatment: importance of cell membrane composition investigated by lipidomics, calorimetry and in vitro efficacy. Sci Rep. 2019;9(1):4758.

https://doi.org/10.1038/s41598-019-41188-z DOI: https://doi.org/10.1038/s41598-019-41188-z

Frandsen SK, Vissing M, Gehl J. A comprehensive review of calcium electroporation – a novel cancer treatment modality. Cancers. 2020;12(2):290.

https://doi.org/10.3390/cancers12020290 DOI: https://doi.org/10.3390/cancers12020290

Staresinic B, Jesenko T, Kamensek U, et al. Effect of calcium electroporation on tumour vasculature. Sci Rep. 2018;8(1):9412.

https://doi.org/10.1038/s41598-018-27728-z DOI: https://doi.org/10.1038/s41598-018-27728-z

Hansen EL, Sozer EB, Romeo S, et al. Dose-dependent ATP depletion and cancer cell death following calcium electroporation, relative effect of calcium concentration and electric field strength. PLoS One. 2015;10(4):e0122973.

https://doi.org/10.1371/journal.pone.0122973 DOI: https://doi.org/10.1371/journal.pone.0122973

Falk H, Forde PF, Bay ML, et al. Calcium electroporation induces tumor eradication, long-lasting immunity and cytokine responses in the CT26 colon cancer mouse model. Oncoimmunology. 2017;6(5):e1301332.

https://doi.org/10.1080/2162402X.2017.1301332 DOI: https://doi.org/10.1080/2162402X.2017.1301332

Frandsen SK, Krüger MB, Mangalanathan UM, et al. Normal and malignant cells exhibit differential responses to calcium electroporation. Cancer Res. 2017;77(16):4389–401.

https://doi.org/10.1158/0008-5472.CAN-16-1611 DOI: https://doi.org/10.1158/0008-5472.CAN-16-1611

Ghosh S, Yang R, Duraki D, et al. Plasma membrane channel TRPM4 mediates immunogenic therapy-induced necrosis. Cancer Res. 2023;83(18):3115–30.

https://doi.org/10.1158/0008-5472.CAN-23-0157 DOI: https://doi.org/10.1158/0008-5472.CAN-23-0157

Kroemer G, Galluzzi L, Kepp O, et al. Immunogenic cell death in cancer therapy. Ann Rev Immunol. 2013;31:51–72.

https://doi.org/10.1146/annurev-immunol-032712-100008 DOI: https://doi.org/10.1146/annurev-immunol-032712-100008

Lisec B, Markelc B, Ursic Valentinuzzi K, et al. The effectiveness of calcium electroporation combined with gene electrotransfer of a plasmid encoding IL-12 is tumor type-dependent. Front Immunol. 2023;14:1189960.

https://doi.org/10.3389/fimmu.2023.1189960 DOI: https://doi.org/10.3389/fimmu.2023.1189960

Badoual C, Hans S, Merillon N, et al. PD-1-expressing tumor-infiltrating T cells are a favorable prognostic biomarker in HPV-associated head and neck cancer. Cancer Res. 2013;73(1):128–38.

https://doi.org/10.1158/0008-5472.CAN-12-2606 DOI: https://doi.org/10.1158/0008-5472.CAN-12-2606

Falk H, Lambaa S, Johannesen HH, et al. Electrochemotherapy and calcium electroporation inducing a systemic immune response with local and distant remission of tumors in a patient with malignant melanoma – a case report. Acta Oncol. 2017;56(8):1126–31.

https://doi.org/10.1080/0284186X.2017.1290274 DOI: https://doi.org/10.1080/0284186X.2017.1290274

Falk H, Matthiessen LW, Wooler G, et al. Calcium electroporation for treatment of cutaneous metastases; a randomized double-blinded phase II study, comparing the effect of calcium electroporation with electrochemotherapy. Acta Oncol. 2018;57(3):311–19.

https://doi.org/10.1080/0284186X.2017.1355109 DOI: https://doi.org/10.1080/0284186X.2017.1355109

Plaschke CC, Gehl J, Johannesen HH, et al. Calcium electroporation for recurrent head and neck cancer: a clinical phase I study. Laryngoscope Investig Otolaryngol. 2019;4(1):49–56.

https://doi.org/10.1002/lio2.233 DOI: https://doi.org/10.1002/lio2.233

Ágoston D, Baltás E, Ócsai H, et al. Evaluation of calcium electroporation for the treatment of cutaneous metastases: a double blinded randomised controlled phase II trial. Cancers. 2020;12(1):179.

https://doi.org/10.3390/cancers12010179 DOI: https://doi.org/10.3390/cancers12010179

Jensen KB, Lonkvist CK, Gehl J, et al. Calcium electroporation for management of cutaneous metastases in HER2-positive breast cancer: A case report. Case Rep Dermatol 2022;14(3):330–8.

https://doi.org/10.1159/000526157 DOI: https://doi.org/10.1159/000526157

Teng MW, Ngiow SF, Ribas A, et al. Classifying cancers based on T-cell infiltration and PD-L1. Cancer Res. 2015;75(11):2139–45.

https://doi.org/10.1158/0008-5472.CAN-15-0255 DOI: https://doi.org/10.1158/0008-5472.CAN-15-0255

Schmid P, Adams S, Rugo HS, et al. Atezolizumab and nab-paclitaxel in advanced triple-negative breast cancer. N Engl J Med. 2018;379(22):2108–21.

https://doi.org/10.1056/NEJMoa1809615 DOI: https://doi.org/10.1056/NEJMoa1809615

Herbst RS, Soria JC, Kowanetz M, et al. Predictive correlates of response to the anti-PD-L1 antibody MPDL3280A in cancer patients. Nature. 2014;515(7528):563–7.

https://doi.org/10.1038/nature14011 DOI: https://doi.org/10.1038/nature14011

Taube JM, Klein A, Brahmer JR, et al. Association of PD-1, PD-1 ligands, and other features of the tumor immune microenvironment with response to anti-PD-1 therapy. Clin Cancer Res. 2014;20(19):5064–74.

https://doi.org/10.1158/1078-0432.CCR-13-3271 DOI: https://doi.org/10.1158/1078-0432.CCR-13-3271

Salgado R, Denkert C, Demaria S, et al. The evaluation of tumor-infiltrating lymphocytes (TILs) in breast cancer: recommendations by an International TILs Working Group 2014. Ann Oncol. 2015;26(2):259–71.

https://doi.org/10.1093/annonc/mdu450 DOI: https://doi.org/10.1093/annonc/mdu450

Loi S, Drubay D, Adams S, et al. Tumor-infiltrating lymphocytes and prognosis: a pooled individual patient analysis of early-stage triple-negative breast cancers. J Clin Oncol. 2019;37(7):559–69.

https://doi.org/10.1200/JCO.18.01010 DOI: https://doi.org/10.1200/JCO.18.01010

Denkert C, von Minckwitz G, Darb-Esfahani S, et al. Tumour-infiltrating lymphocytes and prognosis in different subtypes of breast cancer: a pooled analysis of 3771 patients treated with neoadjuvant therapy. Lancet Oncol. 2018;19(1):40–50.

https://doi.org/10.1016/S1470-2045(17)30904-X DOI: https://doi.org/10.1016/S1470-2045(17)30904-X

Vissing M, Ploen J, Pervan M, et al. Study protocol designed to investigate tumour response to calcium electroporation in cancers affecting the skin: a non-randomised phase II clinical trial. BMJ Open. 2021;11(6):e046779.

https://doi.org/10.1136/bmjopen-2020-046779 DOI: https://doi.org/10.1136/bmjopen-2020-046779

Allison KH, Hammond MEH, Dowsett M, et al. Estrogen and progesterone receptor testing in breast cancer: ASCO/CAP guideline update. J Clin Oncol. 2020;38(12):1346–6.

https://doi.org/10.1200/JCO.19.02309 DOI: https://doi.org/10.1200/JCO.19.02309

Wolff AC, Hammond ME, Hicks DG, et al. 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. Arch Pathol Lab Med. 2014;138(2):241–56.

https://doi.org/10.5858/arpa.2013-0953-SA DOI: https://doi.org/10.5858/arpa.2013-0953-SA

Gundersen HJG. Notes on the estimation of the numerical density of arbitrary profiles: the edge effect. J Microsc. 1977;111(2):219–23.

https://doi.org/10.1111/j.1365-2818.1977.tb00062.x DOI: https://doi.org/10.1111/j.1365-2818.1977.tb00062.x

Vissing M, Munch Larsen L, Christensen AJ, et al. Calcium electroporation as an adjunct therpay for pembrolizumab-resistant bladder cancer: a case report. Acta Oncol. 2023;62(6):614–20.

https://doi.org/10.1080/0284186X.2023.2226330 DOI: https://doi.org/10.1080/0284186X.2023.2226330

Vesely MD, Kershaw MH, Schreiber RD, et al. Natural innate and adaptive immunity to cancer. Ann Rev Immunol. 2011;29:235–71.

https://doi.org/10.1146/annurev-immunol-031210-101324 DOI: https://doi.org/10.1146/annurev-immunol-031210-101324

Gajewski TF, Woo SR, Zha Y, et al. Cancer immunotherapy strategies based on overcoming barriers within the tumor microenvironment. Curr Opin Immunol. 2013;25(2):268–76.

https://doi.org/10.1016/j.coi.2013.02.009 DOI: https://doi.org/10.1016/j.coi.2013.02.009

Fontenot JD, Gavin MA, Rudensky AY. Foxp3 programs the development and function of CD4+CD25+ regulatory T cells. Nat Immunol. 2003;4(4):330–6.

https://doi.org/10.1038/ni904 DOI: https://doi.org/10.1038/ni904

Shevach EM. Mechanisms of foxp3+ T regulatory cell-mediated suppression. Immunity. 2009;30(5):636–45.

https://doi.org/10.1016/j.immuni.2009.04.010 DOI: https://doi.org/10.1016/j.immuni.2009.04.010

Tanaka A, Sakaguchi S. Regulatory T cells in cancer immunotherapy. Cell Res. 2017;27(1):109–18.

https://doi.org/10.1038/cr.2016.151 DOI: https://doi.org/10.1038/cr.2016.151

Galluzzi L, Buqué A, Kepp O, et al. Immunological effects of conventional chemotherapy and targeted anticancer agents. Cancer Cell. 2015;28(6):690–714.

https://doi.org/10.1016/j.ccell.2015.10.012 DOI: https://doi.org/10.1016/j.ccell.2015.10.012

Hegde PS, Karanikas V, Evers S. The where, the when, and the how of immune monitoring for cancer immunotherapies in the era of checkpoint inhibition. Clin Cancer Res. 2016;22(8):1865–74.

https://doi.org/10.1158/1078-0432.CCR-15-1507 DOI: https://doi.org/10.1158/1078-0432.CCR-15-1507

Wu M, Huang Q, Xie Y, et al. Improvement of the anticancer efficacy of PD-1/PD-L1 blockade via combination therapy and PD-L1 regulation. J Hematol Oncol. 2022;15(1):24.

https://doi.org/10.1186/s13045-022-01242-2 DOI: https://doi.org/10.1186/s13045-022-01242-2

Demaria S, Formenti SC. Role of T lymphocytes in tumor response to radiotherapy. Front Oncol. 2012;2:95.

https://doi.org/10.3389/fonc.2012.00095 DOI: https://doi.org/10.3389/fonc.2012.00095

Vissing M, Pervan M, Pløen J, et al. Calcium electroporation in cutaneous metastases – a non-randomised phase II multicentre clinical trial. Eur J Surgical Oncol. 2023;49(9):106925.

https://doi.org/10.1016/j.ejso.2023.04.024 DOI: https://doi.org/10.1016/j.ejso.2023.04.024

Downloads

Additional Files

Published

2024-05-28

How to Cite

Vissing, M., Sinius Pouplier, S., Munch Larsen, L., Krog Frandsen, S., Lodin, A., Lænkholm, A.-V., & Gehl, J. (2024). Immune cell populations in the tumour environment following calcium electropora­tion for cutaneous metastasis: a histopathological study. Acta Oncologica, 63(1), 398–410. https://doi.org/10.2340/1651-226X.2024.19462

Issue

Vol. 63 (2024): Acta Oncologica

Section

Original article

License

Copyright (c) 2023 Mille Vissing, Sandra Sinius Pouplier, Lars Munch Larsen, Stine Krog Frandsen, Alexey Lodin, Anne-Vibeke Lænkholm, Julie Gehl

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

Funding data