IL-1 receptor–associated kinase-3 acts as an immune checkpoint in myeloid cells to limit cancer immunotherapy
Gürcan Tunalı, … , Irineos Papakyriacou, Yumeng Mao
Gürcan Tunalı, … , Irineos Papakyriacou, Yumeng Mao
Published February 9, 2023
Citation Information: J Clin Invest. 2023;133(7):e161084. https://doi.org/10.1172/JCI161084.
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Research Article Immunology Oncology

IL-1 receptor–associated kinase-3 acts as an immune checkpoint in myeloid cells to limit cancer immunotherapy

Abstract

Inflammatory mediators released by cancer cells promote the induction of immune suppression and tolerance in myeloid cells. IL-1 receptor–associated kinase-3 (IRAK3) is a pseudokinase that inhibits IL-1/TLR signaling, but its role in patients treated with immune checkpoint blockade (ICB) therapy remains unclear. Using RNA-Seq data from the IMvigor210 trial, we found that tumors with high IRAK3 expressions showed enriched antiinflammatory pathways and worse clinical response to ICB therapy. Upon IRAK3 protein deletion with CRISPR/Cas9, primary human monocytes displayed altered global protein expression and phosphorylation in quantitative proteomics and released more proinflammatory cytokines in response to stimulation. Bone marrow–derived macrophages from an IRAK3 CRISPR KO mouse model demonstrated a proinflammatory phenotype and enhanced sensitivity to TLR agonists compared with WT cells. IRAK3 deficiency delayed the growth of carcinogen-induced and oncogene-driven murine cancer cells and induced enhanced activation in myeloid cells and T cells. Upon ICB treatment, IRAK3-KO mice showed enrichment of TCF1+PD-1+ stem-like memory CD8+ T cells and resulted in superior growth inhibition of immunologically cold tumors in vivo. Altogether, our study demonstrated what we believe to be a novel cancer-driven immune tolerance program controlled by IRAK3 in humans and mice and proposed its suitability as an immunotherapy target.

Authors

Gürcan Tunalı, Marta Rúbies Bedós, Divya Nagarajan, Patrik Fridh, Irineos Papakyriacou, Yumeng Mao

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Figure 5

Deletion of IRAK3 by CRISPR/Cas9 in mice enhances myeloid cell function.

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Deletion of IRAK3 by CRISPR/Cas9 in mice enhances myeloid cell function....
(A) Design of the IRAK3 CRISPR–KO mice and confirmation with DNA sequencing. Bone marrow cells were isolated from age-matched female C57BL/6NTac WT or IRAK3 CRISPR–KO mice and differentiated in the presence of 100 ng/mL rmGM-CSF for 4 days to macrophages (BMM). (B) Expression of IRAK3 mRNA (n = 3) and protein in response to LPS treatment was measured using qPCR (mean ± SD, unpaired t tests, **P < 0.01, ***P < 0.001) or Western blotting (representative blot from 3 biological repeats). A panel of 754 myeloid cell related genes were measured when WT or IRAK3-KO BMM cells (n = 3 of each) were (C) cultured in media or (D) activated with 100 ng/mL LPS for 24 hours. Data was plotted using log2 fold changes and log10 P values, unpaired student t tests. (E) Changes in expression of selected genes were shown, mean ± SD. (F) BMM cells from at least 4 WT or IRAK3-KO mice were seeded in a 96-well plate and activated with increasing concentrations of LPS or Pam3CSK4 and concentrations of soluble mIL6 or mCXCL1 in culture supernatants were shown after 5 hours. Statistical tests were performed using unpaired t tests. *P < 0.05.