MicroRNA-146a regulates immune-related adverse events caused by immune checkpoint inhibitors
Dominik Marschner, … , Robert Zeiser, Natalie Köhler
Dominik Marschner, … , Robert Zeiser, Natalie Köhler
Published March 3, 2020
Citation Information: JCI Insight. 2020;5(6):e132334. https://doi.org/10.1172/jci.insight.132334.
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Research Article Inflammation Oncology

MicroRNA-146a regulates immune-related adverse events caused by immune checkpoint inhibitors

Abstract

Immune checkpoint inhibitor (ICI) therapy has shown a significant benefit in the treatment of a variety of cancer entities. However, immune-related adverse events (irAEs) occur frequently and can lead to ICI treatment termination. MicroRNA-146a (miR-146a) has regulatory functions in immune cells. We observed that mice lacking miR-146a developed markedly more severe irAEs compared with WT mice in several irAE target organs in 2 different murine models. miR-146a–/– mice exhibited increased T cell activation and effector function upon ICI treatment. Moreover, neutrophil numbers in the spleen and the inflamed intestine were highly increased in ICI-treated miR-146a–/– mice. Therapeutic administration of a miR-146a mimic reduced irAE severity. To validate our preclinical findings in patients, we analyzed the effect of a SNP in the MIR146A gene on irAE severity in 167 patients treated with ICIs. We found that the SNP rs2910164 leading to reduced miR-146a expression was associated with an increased risk of developing severe irAEs, reduced progression-free survival, and increased neutrophil counts both at baseline and during ICI therapy. In conclusion, we characterized miR-146a as a molecular target for preventing ICI-mediated autoimmune dysregulation. Furthermore, we identified the MIR146A SNP rs2910164 as a biomarker to predict severe irAE development in ICI-treated patients.

Authors

Dominik Marschner, Martina Falk, Nora Rebeka Javorniczky, Kathrin Hanke-Müller, Justyna Rawluk, Annette Schmitt-Graeff, Federico Simonetta, Eileen Haring, Severin Dicks, Manching Ku, Sandra Duquesne, Konrad Aumann, David Rafei-Shamsabadi, Frank Meiss, Patrick Marschner, Melanie Boerries, Robert S. Negrin, Justus Duyster, Robert Zeiser, Natalie Köhler

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Figure 2

Increased immune activation signature in miR-146a–deficient CD4 T cells of irAE mice.

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Increased immune activation signature in miR-146a–deficient CD4 T cells ...
(A and B) WT or miR-146a–/– mice (n = 2 per group) were treated with low-dose LPS and anti–PD-1/isotype control antibody for 3 weeks before capturing of MACS purified splenic T cells for scRNA-seq using 10× v3.1 Next GEM chemistry. Data were processed, visualized, and analyzed using the Seurat pipeline v3.0 (45, 46). (A) Uniform Manifold Approximation and Projection (UMAP) plots showing distinct T cell clusters in both miR-146a–/– and WT mice. (B) Gene set enrichment analysis of major T cell clusters. Bivariate heatmap depicts normalized enrichment score as color code and –log10 of the adjusted P value as dot size. Hallmark gene sets were derived from MSigDB. (C and D) WT or miR-146a–/– mice (n = 9–10 per group) were treated with LPS and anti–PD-1/isotype control antibody as indicated. Spleens were isolated on day 22 and CD4+ T cells analyzed by flow cytometry to differentiate naive T cells (CD44–CD62L+), central memory T cells (TCM, CD44+CD62L+, effector memory T cells (TEM, CD44+CD62L–), and activated T cells (CD69+). ***P < 0.001 by 1-way ANOVA followed by Tukey’s post hoc test. Box-and-whisker plot (C) depicts the 25th and the 75th percentiles as the bounds of the boxes, the median as the line within the box, and minimum to maximum as the whiskers. (D) Representative flow cytometry plots showing intracellular IFN-γ staining gated on CD4+ T cells. (E and F) WT or miR-146a–/– mice (n = 9–10 per group) were treated with LPS and anti–PD-1/isotype control antibody as indicated. Splenic CD4+ T cells were assessed by flow cytometry on day 22. (E) Pooled data from 2 independent in vivo experiments. Statistical significance was analyzed by 1-way ANOVA followed by Tukey’s post hoc test. miR-146a, microRNA-146a, irAE, immune-related adverse event, PD-1, programmed cell death protein-1, scRNA-seq, single cell RNA sequencing.