Therapeutic depletion of CCR8+ tumor-infiltrating regulatory T cells elicits antitumor immunity and synergizes with anti-PD-1 therapy
Background Modulation and depletion strategies of regulatory T cells (Tregs) constitute valid approaches in antitumor immunotherapy but suffer from severe adverse effects due to their lack of selectivity for the tumor-infiltrating (ti-)Treg population, indicating the need for a ti-Treg specific biom...
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BMJ Publishing Group
2021-02-01
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| Series: | Journal for ImmunoTherapy of Cancer |
| Online Access: | https://jitc.bmj.com/content/9/2/e001749.full |
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| author | James Dooley Bart Neyns Elizabeth Allen Frank Tacke Louis Boon Damya Laoui Julia Katharina Schwarze Helena Van Damme Bruno Dombrecht Máté Kiss Heleen Roose Eva Van Overmeire Daliya Kancheva Liesbet Martens Aleksandar Murgaski Pauline Madeleine Rachel Bardet Gillian Blancke Maude Jans Evangelia Bolli Maria Solange Martins Yvon Elkrim Kiavash Movahedi Niels Vandamme Sebahat Ocak Isabelle Scheyltjens Lars Vereecke Frank Aboubakar Nana Pascal Merchiers Jo Agnes Van Ginderachter |
| author_facet | James Dooley Bart Neyns Elizabeth Allen Frank Tacke Louis Boon Damya Laoui Julia Katharina Schwarze Helena Van Damme Bruno Dombrecht Máté Kiss Heleen Roose Eva Van Overmeire Daliya Kancheva Liesbet Martens Aleksandar Murgaski Pauline Madeleine Rachel Bardet Gillian Blancke Maude Jans Evangelia Bolli Maria Solange Martins Yvon Elkrim Kiavash Movahedi Niels Vandamme Sebahat Ocak Isabelle Scheyltjens Lars Vereecke Frank Aboubakar Nana Pascal Merchiers Jo Agnes Van Ginderachter |
| author_sort | James Dooley |
| collection | DOAJ |
| description | Background Modulation and depletion strategies of regulatory T cells (Tregs) constitute valid approaches in antitumor immunotherapy but suffer from severe adverse effects due to their lack of selectivity for the tumor-infiltrating (ti-)Treg population, indicating the need for a ti-Treg specific biomarker.Methods We employed single-cell RNA-sequencing in a mouse model of non-small cell lung carcinoma (NSCLC) to obtain a comprehensive overview of the tumor-infiltrating T-cell compartment, with a focus on ti-Treg subpopulations. These findings were validated by flow cytometric analysis of both mouse (LLC-OVA, MC38 and B16-OVA) and human (NSCLC and melanoma) tumor samples. We generated two CCR8-specific nanobodies (Nbs) that recognize distinct epitopes on the CCR8 extracellular domain. These Nbs were formulated as tetravalent Nb-Fc fusion proteins for optimal CCR8 binding and blocking, containing either an antibody-dependent cell-mediated cytotoxicity (ADCC)-deficient or an ADCC-prone Fc region. The therapeutic use of these Nb-Fc fusion proteins was evaluated, either as monotherapy or as combination therapy with anti-programmed cell death protein-1 (anti-PD-1), in both the LLC-OVA and MC38 mouse models.Results We were able to discern two ti-Treg populations, one of which is characterized by the unique expression of Ccr8 in conjunction with Treg activation markers. Ccr8 is also expressed by dysfunctional CD4+ and CD8+ T cells, but the CCR8 protein was only prominent on the highly activated and strongly T-cell suppressive ti-Treg subpopulation of mouse and human tumors, with no major CCR8-positivity found on peripheral Tregs. CCR8 expression resulted from TCR-mediated Treg triggering in an NF-κB-dependent fashion, but was not essential for the recruitment, activation nor suppressive capacity of these cells. While treatment of tumor-bearing mice with a blocking ADCC-deficient Nb-Fc did not influence tumor growth, ADCC-prone Nb-Fc elicited antitumor immunity and reduced tumor growth in synergy with anti-PD-1 therapy. Importantly, ADCC-prone Nb-Fc specifically depleted ti-Tregs in a natural killer (NK) cell-dependent fashion without affecting peripheral Tregs.Conclusions Collectively, our findings highlight the efficacy and safety of targeting CCR8 for the depletion of tumor-promoting ti-Tregs in combination with anti-PD-1 therapy. |
| format | Article |
| id | doaj-art-f3b73327b159432399d5c58c01682dd6 |
| institution | Kabale University |
| issn | 2051-1426 |
| language | English |
| publishDate | 2021-02-01 |
| publisher | BMJ Publishing Group |
| record_format | Article |
| series | Journal for ImmunoTherapy of Cancer |
| spelling | doaj-art-f3b73327b159432399d5c58c01682dd62025-02-02T20:30:09ZengBMJ Publishing GroupJournal for ImmunoTherapy of Cancer2051-14262021-02-019210.1136/jitc-2020-001749Therapeutic depletion of CCR8+ tumor-infiltrating regulatory T cells elicits antitumor immunity and synergizes with anti-PD-1 therapyJames Dooley0Bart Neyns1Elizabeth Allen2Frank Tacke3Louis Boon4Damya Laoui5Julia Katharina Schwarze6Helena Van Damme7Bruno Dombrecht8Máté Kiss9Heleen Roose10Eva Van Overmeire11Daliya Kancheva12Liesbet Martens13Aleksandar Murgaski14Pauline Madeleine Rachel Bardet15Gillian Blancke16Maude Jans17Evangelia Bolli18Maria Solange Martins19Yvon Elkrim20Kiavash Movahedi21Niels Vandamme22Sebahat Ocak23Isabelle Scheyltjens24Lars Vereecke25Frank Aboubakar Nana26Pascal Merchiers27Jo Agnes Van Ginderachter283 Department of Microbiology and Immunology, KU Leuven - University of Leuven, Leuven, BelgiumDepartment of Medical Oncology, University Hospital Brussels, Universitair Ziekenhuis Brussel, Brussels, BelgiumLondon School of Hygiene & Tropical Medicine, London, UKDepartment of Hepatology and Gastroenterology, Charité Universitätsmedizin Berlin, Berlin, GermanyPolpharma Biologics, Utrecht, The NetherlandsLaboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Brussels, Belgium1 Medical Oncology, Universitair Ziekenhuis Brussel, Brussels, BelgiumLaboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Brussels, BelgiumVIB Discovery Sciences, VIB, Ghent, BelgiumLaboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Brussels, Belgium1Aligos Belgium BV, Leuven, Vlaams-Brabant, BelgiumLaboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Brussels, BelgiumLaboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Brussels, BelgiumVIB-UGent Center for Inflammation Research, VIB, Ghent, BelgiumLaboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Brussels, BelgiumLaboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Brussels, BelgiumDepartment of Internal Medicine and Pediatrics, Ghent University, Ghent, BelgiumDepartment of Internal Medicine and Pediatrics, Ghent University, Ghent, BelgiumLaboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Brussels, BelgiumLaboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Brussels, BelgiumMyeloid Cell Immunology Lab, VIB Center for Inflammation Research, Brussels, BelgiumLaboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Brussels, BelgiumData Mining and Modelling for Biomedicine, VIB-UGent Center for Inflammation Research, Ghent, BelgiumInstitut de Recherche Expérimentale et Clinique (IREC), Pôle de Pneumologie, ORL et Dermatologie (PNEU), UCLouvain, Louvain-la-Neuve, BelgiumLaboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Brussels, BelgiumDepartment of Internal Medicine and Pediatrics, Ghent University, Ghent, BelgiumDivision of Pneumology, CHU UCL Namur, Yvoir, Namur, BelgiumOncurious NV, Leuven, BelgiumLaboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Brussels, BelgiumBackground Modulation and depletion strategies of regulatory T cells (Tregs) constitute valid approaches in antitumor immunotherapy but suffer from severe adverse effects due to their lack of selectivity for the tumor-infiltrating (ti-)Treg population, indicating the need for a ti-Treg specific biomarker.Methods We employed single-cell RNA-sequencing in a mouse model of non-small cell lung carcinoma (NSCLC) to obtain a comprehensive overview of the tumor-infiltrating T-cell compartment, with a focus on ti-Treg subpopulations. These findings were validated by flow cytometric analysis of both mouse (LLC-OVA, MC38 and B16-OVA) and human (NSCLC and melanoma) tumor samples. We generated two CCR8-specific nanobodies (Nbs) that recognize distinct epitopes on the CCR8 extracellular domain. These Nbs were formulated as tetravalent Nb-Fc fusion proteins for optimal CCR8 binding and blocking, containing either an antibody-dependent cell-mediated cytotoxicity (ADCC)-deficient or an ADCC-prone Fc region. The therapeutic use of these Nb-Fc fusion proteins was evaluated, either as monotherapy or as combination therapy with anti-programmed cell death protein-1 (anti-PD-1), in both the LLC-OVA and MC38 mouse models.Results We were able to discern two ti-Treg populations, one of which is characterized by the unique expression of Ccr8 in conjunction with Treg activation markers. Ccr8 is also expressed by dysfunctional CD4+ and CD8+ T cells, but the CCR8 protein was only prominent on the highly activated and strongly T-cell suppressive ti-Treg subpopulation of mouse and human tumors, with no major CCR8-positivity found on peripheral Tregs. CCR8 expression resulted from TCR-mediated Treg triggering in an NF-κB-dependent fashion, but was not essential for the recruitment, activation nor suppressive capacity of these cells. While treatment of tumor-bearing mice with a blocking ADCC-deficient Nb-Fc did not influence tumor growth, ADCC-prone Nb-Fc elicited antitumor immunity and reduced tumor growth in synergy with anti-PD-1 therapy. Importantly, ADCC-prone Nb-Fc specifically depleted ti-Tregs in a natural killer (NK) cell-dependent fashion without affecting peripheral Tregs.Conclusions Collectively, our findings highlight the efficacy and safety of targeting CCR8 for the depletion of tumor-promoting ti-Tregs in combination with anti-PD-1 therapy.https://jitc.bmj.com/content/9/2/e001749.full |
| spellingShingle | James Dooley Bart Neyns Elizabeth Allen Frank Tacke Louis Boon Damya Laoui Julia Katharina Schwarze Helena Van Damme Bruno Dombrecht Máté Kiss Heleen Roose Eva Van Overmeire Daliya Kancheva Liesbet Martens Aleksandar Murgaski Pauline Madeleine Rachel Bardet Gillian Blancke Maude Jans Evangelia Bolli Maria Solange Martins Yvon Elkrim Kiavash Movahedi Niels Vandamme Sebahat Ocak Isabelle Scheyltjens Lars Vereecke Frank Aboubakar Nana Pascal Merchiers Jo Agnes Van Ginderachter Therapeutic depletion of CCR8+ tumor-infiltrating regulatory T cells elicits antitumor immunity and synergizes with anti-PD-1 therapy Journal for ImmunoTherapy of Cancer |
| title | Therapeutic depletion of CCR8+ tumor-infiltrating regulatory T cells elicits antitumor immunity and synergizes with anti-PD-1 therapy |
| title_full | Therapeutic depletion of CCR8+ tumor-infiltrating regulatory T cells elicits antitumor immunity and synergizes with anti-PD-1 therapy |
| title_fullStr | Therapeutic depletion of CCR8+ tumor-infiltrating regulatory T cells elicits antitumor immunity and synergizes with anti-PD-1 therapy |
| title_full_unstemmed | Therapeutic depletion of CCR8+ tumor-infiltrating regulatory T cells elicits antitumor immunity and synergizes with anti-PD-1 therapy |
| title_short | Therapeutic depletion of CCR8+ tumor-infiltrating regulatory T cells elicits antitumor immunity and synergizes with anti-PD-1 therapy |
| title_sort | therapeutic depletion of ccr8 tumor infiltrating regulatory t cells elicits antitumor immunity and synergizes with anti pd 1 therapy |
| url | https://jitc.bmj.com/content/9/2/e001749.full |
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