Single-cell characterization of anti–LAG-3 and anti–PD-1 combination treatment in patients with melanoma
Jani Huuhtanen, Henna Kasanen, Katriina Peltola, Tapio Lönnberg, Virpi Glumoff, Oscar Brück, Olli Dufva, Karita Peltonen, Johanna Vikkula, Emmi Jokinen, Mette Ilander, Moon Hee Lee, Siru Mäkelä, Marta Nyakas, Bin Li, Micaela Hernberg, Petri Bono, Harri Lähdesmäki, Anna Kreutzman, Satu Mustjoki
Jani Huuhtanen, Henna Kasanen, Katriina Peltola, Tapio Lönnberg, Virpi Glumoff, Oscar Brück, Olli Dufva, Karita Peltonen, Johanna Vikkula, Emmi Jokinen, Mette Ilander, Moon Hee Lee, Siru Mäkelä, Marta Nyakas, Bin Li, Micaela Hernberg, Petri Bono, Harri Lähdesmäki, Anna Kreutzman, Satu Mustjoki
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Clinical Research and Public Health Immunology Oncology

Single-cell characterization of anti–LAG-3 and anti–PD-1 combination treatment in patients with melanoma

Abstract

Background Relatlimab plus nivolumab (anti–lymphocyte-activation gene 3 plus anti–programmed death 1 [anti–LAG-3+anti–PD-1]) has been approved by the FDA as a first-line therapy for stage III/IV melanoma, but its detailed effect on the immune system is unknown.Methods We evaluated blood samples from 40 immunotherapy-naive or prior immunotherapy–refractory patients with metastatic melanoma treated with anti–LAG-3+anti–PD-1 in a phase I trial using single-cell RNA and T cell receptor sequencing (scRNA+TCRαβ-Seq) combined with other multiomics profiling.Results The highest LAG3 expression was noted in NK cells, Tregs, and CD8+ T cells, and these cell populations underwent the most significant changes during the treatment. Adaptive NK cells were enriched in responders and underwent profound transcriptomic changes during the therapy, resulting in an active phenotype. LAG3+ Tregs expanded, but based on the transcriptome profile, became metabolically silent during the treatment. Last, higher baseline TCR clonality was observed in responding patients, and their expanding CD8+ T cell clones gained a more cytotoxic and NK-like phenotype.Conclusion Anti–LAG-3+anti–PD-1 therapy has profound effects on NK cells and Tregs in addition to CD8+ T cells.Trial registration ClinicalTrials.gov (NCT01968109)Funding Cancer Foundation Finland, Sigrid Juselius Foundation, Signe and Ane Gyllenberg Foundation, Relander Foundation, State funding for university-level health research in Finland, a Helsinki Institute of Life Sciences Fellow grant, Academy of Finland (grant numbers 314442, 311081, 335432, and 335436), and an investigator-initiated research grant from BMS.

Authors

Jani Huuhtanen, Henna Kasanen, Katriina Peltola, Tapio Lönnberg, Virpi Glumoff, Oscar Brück, Olli Dufva, Karita Peltonen, Johanna Vikkula, Emmi Jokinen, Mette Ilander, Moon Hee Lee, Siru Mäkelä, Marta Nyakas, Bin Li, Micaela Hernberg, Petri Bono, Harri Lähdesmäki, Anna Kreutzman, Satu Mustjoki

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Figure 4

Anti–LAG-3+anti–PD-1 enhances NK cell degranulation, cytokine secretion, and T cell proliferation.

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Anti–LAG-3+anti–PD-1 enhances NK cell degranulation, cytokine secretion,...
(A) Degranulation (CD107a/b) and IFN-γ/TNF-α production of primary NK cells after stimulation with the chronic myelogenous leukemia (CML) cell line K562 in patients before (up in pre) and after (up in post) anti–LAG-3+anti–PD-1 treatment and in healthy donors at different time points (n = 4, 12 samples). When samples were available, they were examined as 3 replicates, but here, values are shown as averages. P values were calculated with the Kruskal-Wallis test. (B) Relationship between extracellular LAG-3 and degranulation responses (CD107a/b) to K562 target cells in melanoma samples before and after anti–LAG-3+anti–PD-1 treatment (n = 10, 30 samples). P values and correlation coefficients were calculated with Spearman’s rank correlation. (C) Top: Proliferation of CD4+ and CD8+ T cells induced by anti-CD3 and -CD28 beads (n = 3, 9 samples). Bottom: Cell divisions in CD4+ and CD8+ T cells after anti-CD3 and -CD28 bead stimulation in a selected patient. Cell proliferation was traced with flow cytometry by dilution of CellTrace Violet dye. Cells from different time points (before and after therapy) are marked with different colors.