1. Therapeutic depletion of CCR8 + tumor-infiltrating regulatory T cells elicits antitumor immunity and synergizes with anti-PD-1 therapy.
- Author
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Van Damme H, Dombrecht B, Kiss M, Roose H, Allen E, Van Overmeire E, Kancheva D, Martens L, Murgaski A, Bardet PMR, Blancke G, Jans M, Bolli E, Martins MS, Elkrim Y, Dooley J, Boon L, Schwarze JK, Tacke F, Movahedi K, Vandamme N, Neyns B, Ocak S, Scheyltjens I, Vereecke L, Nana FA, Merchiers P, Laoui D, and Van Ginderachter JA
- Subjects
- Animals, Carcinoma, Lewis Lung genetics, Carcinoma, Lewis Lung immunology, Carcinoma, Lewis Lung metabolism, Combined Modality Therapy, Databases, Genetic, Female, Gene Expression Profiling, Humans, Lung Neoplasms genetics, Lung Neoplasms immunology, Lung Neoplasms metabolism, Lymphocytes, Tumor-Infiltrating metabolism, Melanoma, Experimental genetics, Melanoma, Experimental immunology, Melanoma, Experimental metabolism, Mice, Inbred C57BL, Mice, Knockout, Molecular Targeted Therapy, Phenotype, Programmed Cell Death 1 Receptor metabolism, RNA-Seq, Receptors, CCR8 genetics, Skin Neoplasms genetics, Skin Neoplasms immunology, Skin Neoplasms metabolism, T-Lymphocytes, Regulatory metabolism, Mice, Antineoplastic Agents, Immunological pharmacology, Carcinoma, Lewis Lung therapy, Immune Checkpoint Inhibitors pharmacology, Lung Neoplasms drug therapy, Lymphocyte Depletion, Lymphocytes, Tumor-Infiltrating immunology, Melanoma, Experimental therapy, Programmed Cell Death 1 Receptor antagonists & inhibitors, Receptors, CCR8 deficiency, Skin Neoplasms therapy, T-Lymphocytes, Regulatory immunology
- Abstract
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., Competing Interests: Competing interests: HR and EA are employees of Oncurious NV. PM is a consultant to Oncurious NV., (© Author(s) (or their employer(s)) 2021. Re-use permitted under CC BY. Published by BMJ.)- Published
- 2021
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