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3. Immune-based biomarker accurately predicts response to imiquimod immunotherapy in cervical high-grade squamous intraepithelial lesions

Author

Abdulrahman, Ziena, primary, Hendriks, Natasja, additional, J Kruse, Arnold, additional, Somarakis, Antonios, additional, J M van de Sande, Anna, additional, J van Beekhuizen, Heleen, additional, M J Piek, Jurgen, additional, de Miranda, Noel F C C, additional, Kooreman, Loes F S, additional, F M Slangen, Brigitte, additional, van der Burg, Sjoerd H, additional, de Vos van Steenwijk, Peggy J, additional, and van Esch, Edith M G, additional

Published
2022
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4. Immune-based biomarker accurately predicts response to imiquimod immunotherapy in cervical high-grade squamous intraepithelial lesions

Author

Abdulrahman, Ziena, Hendriks, Natasja, J Kruse, Arnold, Somarakis, Antonios, J M van de Sande, Anna, J van Beekhuizen, Heleen, M J Piek, Jurgen, de Miranda, Noel F.C.C., Kooreman, Loes F.S., F M Slangen, Brigitte, van der Burg, Sjoerd H., de Vos van Steenwijk, Peggy J., van Esch, Edith M.G., Abdulrahman, Ziena, Hendriks, Natasja, J Kruse, Arnold, Somarakis, Antonios, J M van de Sande, Anna, J van Beekhuizen, Heleen, M J Piek, Jurgen, de Miranda, Noel F.C.C., Kooreman, Loes F.S., F M Slangen, Brigitte, van der Burg, Sjoerd H., de Vos van Steenwijk, Peggy J., and van Esch, Edith M.G.

Abstract

BACKGROUND: The complete response rate of cervical high-grade squamous intraepithelial lesion (cHSIL) patients to imiquimod immunotherapy is approximately 60%. Consequently, many patients are exposed to unnecessary adverse effects of imiquimod. On the other hand, conventional surgical large loop excision therapy is associated with increased risk of premature births in subsequent pregnancies. An in-depth analysis of the cHSIL immune microenvironment was performed in order to identify and develop a predictive biomarker for response to imiquimod, to maximize therapy efficacy and to avoid adverse effects in patients unlikely to respond. METHODS: Biopsies of 35 cHSIL patients, before and 10 weeks on imiquimod treatment, were analyzed by two multispectral seven-color immunofluorescence panels for T cell and myeloid cell composition in relation to treatment response. Based on these results a simplified immunohistochemical detection protocol was developed. Samples were scanned with the Vectra multispectral imaging system and cells were automatically identified using machine learning. RESULTS: The immune microenvironment of complete responders (CR) is characterized by a strong and coordinated infiltration by T helper cells (activated PD1+/type 1 Tbet+), M1-like macrophages (CD68+CD163-) and dendritic cells (CD11c+) prior to imiquimod. The lesions of non-responders (NRs) displayed a high infiltration by CD3+FOXP3+ regulatory T cells. At 10 weeks on imiquimod, a strong influx of intraepithelial and stromal CD4+ T cells was observed in CR but not NR patients. A steep decrease in macrophages occurred both in CR and NR patients, leveling the pre-existing differences in myeloid cell composition between the two groups. Based on the pre-existing immune composition differences, the sum of intraepithelial CD4 T cell, macrophage and dendritic cell counts was used to develop a quantitative simplified one color immunohistochemical biomarker, the CHSIL immune biomarker for imiquimod (C

Published
2022

5. Tumor-specific T cells support chemokine-driven spatial organization of intratumoral immune microaggregates needed for long survival

Author

Abdulrahman, Ziena, primary, Santegoets, Saskia J, additional, Sturm, Gregor, additional, Charoentong, Pornpimol, additional, Ijsselsteijn, Marieke E, additional, Somarakis, Antonios, additional, Höllt, Thomas, additional, Finotello, Francesca, additional, Trajanoski, Zlatko, additional, van Egmond, Sylvia L, additional, Mustafa, Dana A M, additional, Welters, Marij J P, additional, de Miranda, Noel F C C, additional, and van der Burg, Sjoerd H, additional

Published
2022
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6. CD161 expression and regulation defines rapidly responding effector CD4+ T cells associated with improved survival in HPV16-associated tumors

Author

Duurland, Chantal L, primary, Santegoets, Saskia J, additional, Abdulrahman, Ziena, additional, Loof, Nikki M, additional, Sturm, Gregor, additional, Wesselink, Tom H, additional, Arens, Ramon, additional, Boekestijn, Sanne, additional, Ehsan, Ilina, additional, van Poelgeest, Mariette I E, additional, Finotello, Francesca, additional, Hackl, Hubert, additional, Trajanoski, Zlatko, additional, ten Dijke, Peter, additional, Braud, Veronique M, additional, Welters, Marij J P, additional, and van der Burg, Sjoerd H, additional

Published
2022
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7. 35 Chemokine-driven spatial organization of immune cell microaggregates marks oropharyngeal squamous cell carcinomas containing tumor-specific T cells

Author

Abdulrahman, Ziena, primary, Santegoets, Saskia, additional, Sturm, Gregor, additional, Charoentong, Pornpimol, additional, Ijsselsteijn, Marieke, additional, Somarakis, Antonios, additional, Höllt, Thomas, additional, Finotello, Francesca, additional, Trajanoski, Zlatko, additional, Egmond, Sylvia van, additional, Mustafa, Dana, additional, Welters, Marij, additional, Miranda, Noel de, additional, and Burg, Sjoerd van der, additional

Published
2021
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8. Cancer immunophenotyping by seven-colour multispectral imaging without tyramide signal amplification

Author

Ijsselsteijn, Marieke E, Brouwer, Thomas P, Abdulrahman, Ziena, Reidy, Eileen, Ramalheiro, Ana, Heeren, A Marijne, Vahrmeijer, Alexander, Jordanova, Ekaterina S, de Miranda, Noel FCC, CCA - Cancer biology and immunology, AII - Cancer immunology, Amsterdam Reproduction & Development (AR&D), Neurology, and Obstetrics and Gynaecology

Subjects
immune infiltration, Sequence Analysis, RNA, T-cells, biomarkers, Fluorescent Antibody Technique, Original Articles, multispectral immunofluorescence, T‐cells, Immunophenotyping, Lymphocytes, Tumor-Infiltrating, myeloid cells, Neoplasms, Biomarkers, Tumor, Humans, Original Article, Immunotherapy, cancer microenvironment
Abstract

Checkpoint blockade immunotherapies have revolutionised cancer treatment in the last decade. Nevertheless, these are only beneficial for a small proportion of cancer patients. Important prognosticators for response to immunotherapy are the mutation burden of tumours as well as the quality and quantity of tumour‐infiltrating immune cells. High‐throughput multiplex immunophenotyping technologies have a central role in deciphering the complexity of anti‐tumour immune responses. Current techniques for the immunophenotyping of solid tumours are held back by the lack of spatial context, limitations in the number of targets that can be visualised simultaneously, and/or cumbersome protocols. We developed a tyramide signal amplification‐free method for the simultaneous detection of seven cellular targets by immunofluorescence. This method overcomes limitations posed by most widespread techniques and provides a unique tool for extensive phenotyping by multispectral fluorescence microscopy. Furthermore, it can be easily implemented as a high‐throughput technology for validation of discovery sets generated by RNA sequencing or mass cytometry and may serve in the future as a complementary diagnostic tool.

Published
2019

9. Lack of myeloid cell infiltration as an acquired resistance strategy to immunotherapy

Author

Beyranvand Nejad, Elham, primary, Labrie, Camilla, additional, Abdulrahman, Ziena, additional, van Elsas, Marit J, additional, Rademaker, Eva, additional, Kleinovink, Jan Willem, additional, van der Sluis, Tetje C, additional, van Duikeren, Suzanne, additional, Teunisse, Amina F A.S, additional, Jochemsen, Aart G, additional, Oosting, Jan, additional, de Miranda, Noel F C C, additional, Van Hall, Thorbald, additional, Arens, Ramon, additional, and van der Burg, Sjoerd H, additional

Published
2020
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11. Cancer immunophenotyping by 7 colour multispectral imaging without tyramide signal amplification

Author

Ijselstein, Marieke, Brouwer, Thomas, Abdulrahman, Ziena, Reidy, Eileen, Ramalheiro, Ana, Heeren, AM, Vahrmeijer, Alexander, Jordanova, ES, de Miranda, Noel, Medical oncology laboratory, CCA - Cancer biology and immunology, AII - Cancer immunology, Obstetrics and gynaecology, and Amsterdam Reproduction & Development (AR&D)

Abstract

Checkpoint blockade immunotherapies have revolutionised cancer treatment in the last decade. Nevertheless, these are only beneficial for a small proportion of cancer patients. Important prognosticators for response to immunotherapy are the mutation burden of tumours as well as the quality and quantity of tumour‐infiltrating immune cells. High‐throughput multiplex immunophenotyping technologies have a central role in deciphering the complexity of anti‐tumour immune responses. Current techniques for the immunophenotyping of solid tumours are held back by the lack of spatial context, limitations in the number of targets that can be visualised simultaneously, and/or cumbersome protocols. We developed a tyramide signal amplification (TSA) – free method for the simultaneous detection of 7 cellular targets by immunofluorescence. This method overcomes limitations posed by most widespread techniques and provides a unique tool for extensive phenotyping by multispectral fluorescence microscopy. Furthermore, it can be easily implemented as a high‐throughput technology for validation of discovery sets generated by RNA sequencing or mass cytometry and may serve in the future as a complementary diagnostic tool.

Published
2019
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