IFN-α with dasatinib broadens the immune repertoire in patients with chronic-phase chronic myeloid leukemia
Jani Huuhtanen, Mette Ilander, Bhagwan Yadav, Olli M.J. Dufva, Hanna Lähteenmäki, Tiina Kasanen, Jay Klievink, Ulla Olsson-Strömberg, Jesper Stentoft, Johan Richter, Perttu Koskenvesa, Martin Höglund, Stina Söderlund, Arta Dreimane, Kimmo Porkka, Tobias Gedde-Dahl, Björn T. Gjertsen, Leif Stenke, Kristina Myhr-Eriksson, Berit Markevärn, Anna Lübking, Andreja Dimitrijevic, Lene Udby, Ole Weis Bjerrum, Henrik Hjorth-Hansen, Satu Mustjoki
Jani Huuhtanen, Mette Ilander, Bhagwan Yadav, Olli M.J. Dufva, Hanna Lähteenmäki, Tiina Kasanen, Jay Klievink, Ulla Olsson-Strömberg, Jesper Stentoft, Johan Richter, Perttu Koskenvesa, Martin Höglund, Stina Söderlund, Arta Dreimane, Kimmo Porkka, Tobias Gedde-Dahl, Björn T. Gjertsen, Leif Stenke, Kristina Myhr-Eriksson, Berit Markevärn, Anna Lübking, Andreja Dimitrijevic, Lene Udby, Ole Weis Bjerrum, Henrik Hjorth-Hansen, Satu Mustjoki
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Research Article Hematology Immunology

IFN-α with dasatinib broadens the immune repertoire in patients with chronic-phase chronic myeloid leukemia

Abstract

In chronic myeloid leukemia (CML), combination therapies with tyrosine kinase inhibitors (TKIs) aim to improve the achievement of deep molecular remission that would allow therapy discontinuation. IFN-α is one promising candidate, as it has long-lasting effects on both malignant and immune cells. In connection with a multicenter clinical trial combining dasatinib with IFN-α in 40 patients with chronic-phase CML (NordCML007, NCT01725204), we performed immune monitoring with single-cell RNA and T cell receptor (TCR) sequencing (n = 4, 12 samples), bulk TCRβ sequencing (n = 13, 26 samples), flow cytometry (n = 40, 106 samples), cytokine analyses (n = 17, 80 samples), and ex vivo functional studies (n = 39, 80 samples). Dasatinib drove the immune repertoire toward terminally differentiated NK and CD8+ T cells with dampened functional capabilities. Patients with dasatinib-associated pleural effusions had increased numbers of CD8+ recently activated effector memory T (Temra) cells. In vitro, dasatinib prevented CD3-induced cell death by blocking TCR signaling. The addition of IFN-α reversed the terminally differentiated phenotypes and increased the number of costimulatory intercellular interactions and the number of unique putative epitope-specific TCR clusters. In vitro IFN-α had costimulatory effects on TCR signaling. Our work supports the combination of IFN-α with TKI therapy, as IFN-α broadens the immune repertoire and restores immunological function.

Authors

Jani Huuhtanen, Mette Ilander, Bhagwan Yadav, Olli M.J. Dufva, Hanna Lähteenmäki, Tiina Kasanen, Jay Klievink, Ulla Olsson-Strömberg, Jesper Stentoft, Johan Richter, Perttu Koskenvesa, Martin Höglund, Stina Söderlund, Arta Dreimane, Kimmo Porkka, Tobias Gedde-Dahl, Björn T. Gjertsen, Leif Stenke, Kristina Myhr-Eriksson, Berit Markevärn, Anna Lübking, Andreja Dimitrijevic, Lene Udby, Ole Weis Bjerrum, Henrik Hjorth-Hansen, Satu Mustjoki

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Figure 3

The addition of IFN-α reversed the dasatinib-induced maturation of NK cells and CD8+ and CD4+ T cells and restored immunological function.

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The addition of IFN-α reversed the dasatinib-induced maturation of NK ce...
(A) The abundances of different mature populations of NK cells and CD8+ and CD4+ T cells at different time points shown as percentages of given parent populations. The P values were calculated with the Kruskal-Wallis test. (B) UMAP projections of NK cell and CD8+ and CD4+ T cell clusters identified in Figure 1B, where the superimposed lines represent the predicted maturation trajectories. (C) The same as in B showing the cell densities at different time points, where the more mature clusters are replaced by immature clusters after the addition of IFN-α to dasatinib. (D) The proportion of cells belonging to different clusters in individual CD8+ clones, in which the transition toward the IFNG-producing Temra cluster (cluster 9) can be seen (15 of 32 [46.88%] clones with at least 5 cells). (E) The proportion of CD8+ cells belonging to cluster 9 from the 32 clones with at least 5 cells at different time points. The P value was calculated with the Mann-Whitney test. (F) Cell type abundances of degranulating (CD107+) and IFN-γ/TNF-α–producing T cells after being stimulated with anti-CD3, anti-CD28, and anti-CD49d. The P values were calculated with the Kruskal-Wallis test. (G) The amount of TCR NFAT activity (measured by luciferase) and cell viability (measured by CellTiter Glo assay, normalized to unstimulated wells) after being stimulated with anti-CD3, anti-CD28, and IFN-α in the presence of DMSO or dasatinib. The P values were calculated with the 2-sided Kruskal-Wallis test. *P < 0.05; **P < 0.01. NS, not significant. Box-and-whisker plots are defined in the Methods.