Flow Cytometric Assessment of FcγRIIIa-V158F Polymorphisms and NK Cell Mediated ADCC Revealed Reduced NK Cell Functionality in Colorectal Cancer Patients
Antibody-dependent cell-mediated cytotoxicity (ADCC) by NK cells is a key mechanism in anti-cancer therapies with monoclonal antibodies, including cetuximab (EGFR-targeting) and avelumab (PDL1-targeting). Fc gamma receptor IIIa (FcγRIIIa) polymorphisms impact ADCC, yet their clinical relevance in NK...
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2024-12-01
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| author | Phillip Schiele Stefan Kolling Stanislav Rosnev Charlotte Junkuhn Anna Luzie Walter Jobst Christian von Einem Sebastian Stintzing Wenzel Schöning Igor Maximilian Sauer Dominik Paul Modest Kathrin Heinrich Lena Weiss Volker Heinemann Lars Bullinger Marco Frentsch Il-Kang Na |
| author_facet | Phillip Schiele Stefan Kolling Stanislav Rosnev Charlotte Junkuhn Anna Luzie Walter Jobst Christian von Einem Sebastian Stintzing Wenzel Schöning Igor Maximilian Sauer Dominik Paul Modest Kathrin Heinrich Lena Weiss Volker Heinemann Lars Bullinger Marco Frentsch Il-Kang Na |
| author_sort | Phillip Schiele |
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| description | Antibody-dependent cell-mediated cytotoxicity (ADCC) by NK cells is a key mechanism in anti-cancer therapies with monoclonal antibodies, including cetuximab (EGFR-targeting) and avelumab (PDL1-targeting). Fc gamma receptor IIIa (FcγRIIIa) polymorphisms impact ADCC, yet their clinical relevance in NK cell functionality remains debated. We developed two complementary flow cytometry assays: one to predict the FcγRIIIa-V158F polymorphism using a machine learning model, and a 15-color flow cytometry panel to assess antibody-induced NK cell functionality and cancer-immune cell interactions. Samples were collected from healthy donors and metastatic colorectal cancer (mCRC) patients from the FIRE-6-Avelumab phase II study. The machine learning model accurately predicted the FcγRIIIa-V158F polymorphism in 94% of samples. FF homozygous patients showed diminished cetuximab-mediated ADCC compared to VF or VV carriers. In mCRC patients, NK cell dysfunctions were evident as impaired ADCC, decreased CD16 downregulation, and reduced CD137/CD107a induction. Elevated PD1+ NK cell levels, reduced lysis of PDL1-expressing CRC cells and improved NK cell activation in combination with the PDL1-targeting avelumab indicate that the PD1-PDL1 axis contributes to impaired cetuximab-induced NK cell function. Together, these optimized assays effectively identify NK cell dysfunctions in mCRC patients and offer potential for broader application in evaluating NK cell functionality across cancers and therapeutic settings. |
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| institution | Kabale University |
| issn | 2073-4409 |
| language | English |
| publishDate | 2024-12-01 |
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| spelling | doaj-art-b10cac1d33f040a9869cba77c08895b52025-01-10T13:16:18ZengMDPI AGCells2073-44092024-12-011413210.3390/cells14010032Flow Cytometric Assessment of FcγRIIIa-V158F Polymorphisms and NK Cell Mediated ADCC Revealed Reduced NK Cell Functionality in Colorectal Cancer PatientsPhillip Schiele0Stefan Kolling1Stanislav Rosnev2Charlotte Junkuhn3Anna Luzie Walter4Jobst Christian von Einem5Sebastian Stintzing6Wenzel Schöning7Igor Maximilian Sauer8Dominik Paul Modest9Kathrin Heinrich10Lena Weiss11Volker Heinemann12Lars Bullinger13Marco Frentsch14Il-Kang Na15BIH Center for Regenerative Therapies (BCRT), Therapy-Induced Remodeling in Immuno-Oncology, Berlin Institute of Health at Charité—Universitätsmedizin Berlin, 13353 Berlin, GermanyBIH Center for Regenerative Therapies (BCRT), Therapy-Induced Remodeling in Immuno-Oncology, Berlin Institute of Health at Charité—Universitätsmedizin Berlin, 13353 Berlin, GermanyDepartment of Hematology, Oncology and Cancer Immunology, Charité—Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt—Universität zu Berlin, 13353 Berlin, GermanyBIH Center for Regenerative Therapies (BCRT), Therapy-Induced Remodeling in Immuno-Oncology, Berlin Institute of Health at Charité—Universitätsmedizin Berlin, 13353 Berlin, GermanyBIH Center for Regenerative Therapies (BCRT), Therapy-Induced Remodeling in Immuno-Oncology, Berlin Institute of Health at Charité—Universitätsmedizin Berlin, 13353 Berlin, GermanyDepartment of Hematology, Oncology and Cancer Immunology, Charité—Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt—Universität zu Berlin, 13353 Berlin, GermanyDepartment of Hematology, Oncology and Cancer Immunology, Charité—Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt—Universität zu Berlin, 13353 Berlin, GermanyDepartment of Surgery, Campus Charité Mitte—Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt—Universität zu Berlin, 10117 Berlin, GermanyDepartment of Surgery, Campus Virchow Klinikum, Charité—Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt—Universität zu Berlin, 13353 Berlin, GermanyDepartment of Hematology, Oncology and Cancer Immunology, Charité—Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt—Universität zu Berlin, 13353 Berlin, GermanyDepartment of Medicine III, Ludwig-Maximilians-University of Munich, 80539 Munich, GermanyDepartment of Hematology/Oncology and Comprehensive Cancer Center, University Hospital, Klinikum Grosshadern, Ludwig-Maximilians-University of Munich, 80539 Munich, GermanyDepartment of Hematology/Oncology and Comprehensive Cancer Center, University Hospital, Klinikum Grosshadern, Ludwig-Maximilians-University of Munich, 80539 Munich, GermanyDepartment of Hematology, Oncology and Cancer Immunology, Charité—Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt—Universität zu Berlin, 13353 Berlin, GermanyBIH Center for Regenerative Therapies (BCRT), Therapy-Induced Remodeling in Immuno-Oncology, Berlin Institute of Health at Charité—Universitätsmedizin Berlin, 13353 Berlin, GermanyBIH Center for Regenerative Therapies (BCRT), Therapy-Induced Remodeling in Immuno-Oncology, Berlin Institute of Health at Charité—Universitätsmedizin Berlin, 13353 Berlin, GermanyAntibody-dependent cell-mediated cytotoxicity (ADCC) by NK cells is a key mechanism in anti-cancer therapies with monoclonal antibodies, including cetuximab (EGFR-targeting) and avelumab (PDL1-targeting). Fc gamma receptor IIIa (FcγRIIIa) polymorphisms impact ADCC, yet their clinical relevance in NK cell functionality remains debated. We developed two complementary flow cytometry assays: one to predict the FcγRIIIa-V158F polymorphism using a machine learning model, and a 15-color flow cytometry panel to assess antibody-induced NK cell functionality and cancer-immune cell interactions. Samples were collected from healthy donors and metastatic colorectal cancer (mCRC) patients from the FIRE-6-Avelumab phase II study. The machine learning model accurately predicted the FcγRIIIa-V158F polymorphism in 94% of samples. FF homozygous patients showed diminished cetuximab-mediated ADCC compared to VF or VV carriers. In mCRC patients, NK cell dysfunctions were evident as impaired ADCC, decreased CD16 downregulation, and reduced CD137/CD107a induction. Elevated PD1+ NK cell levels, reduced lysis of PDL1-expressing CRC cells and improved NK cell activation in combination with the PDL1-targeting avelumab indicate that the PD1-PDL1 axis contributes to impaired cetuximab-induced NK cell function. Together, these optimized assays effectively identify NK cell dysfunctions in mCRC patients and offer potential for broader application in evaluating NK cell functionality across cancers and therapeutic settings.https://www.mdpi.com/2073-4409/14/1/32antibody-dependent cell cytotoxicitycetuximabcolorectal neoplasmsflow cytometrysingle nucleotide polymorphismnatural killer cells |
| spellingShingle | Phillip Schiele Stefan Kolling Stanislav Rosnev Charlotte Junkuhn Anna Luzie Walter Jobst Christian von Einem Sebastian Stintzing Wenzel Schöning Igor Maximilian Sauer Dominik Paul Modest Kathrin Heinrich Lena Weiss Volker Heinemann Lars Bullinger Marco Frentsch Il-Kang Na Flow Cytometric Assessment of FcγRIIIa-V158F Polymorphisms and NK Cell Mediated ADCC Revealed Reduced NK Cell Functionality in Colorectal Cancer Patients Cells antibody-dependent cell cytotoxicity cetuximab colorectal neoplasms flow cytometry single nucleotide polymorphism natural killer cells |
| title | Flow Cytometric Assessment of FcγRIIIa-V158F Polymorphisms and NK Cell Mediated ADCC Revealed Reduced NK Cell Functionality in Colorectal Cancer Patients |
| title_full | Flow Cytometric Assessment of FcγRIIIa-V158F Polymorphisms and NK Cell Mediated ADCC Revealed Reduced NK Cell Functionality in Colorectal Cancer Patients |
| title_fullStr | Flow Cytometric Assessment of FcγRIIIa-V158F Polymorphisms and NK Cell Mediated ADCC Revealed Reduced NK Cell Functionality in Colorectal Cancer Patients |
| title_full_unstemmed | Flow Cytometric Assessment of FcγRIIIa-V158F Polymorphisms and NK Cell Mediated ADCC Revealed Reduced NK Cell Functionality in Colorectal Cancer Patients |
| title_short | Flow Cytometric Assessment of FcγRIIIa-V158F Polymorphisms and NK Cell Mediated ADCC Revealed Reduced NK Cell Functionality in Colorectal Cancer Patients |
| title_sort | flow cytometric assessment of fcγriiia v158f polymorphisms and nk cell mediated adcc revealed reduced nk cell functionality in colorectal cancer patients |
| topic | antibody-dependent cell cytotoxicity cetuximab colorectal neoplasms flow cytometry single nucleotide polymorphism natural killer cells |
| url | https://www.mdpi.com/2073-4409/14/1/32 |
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