Blocking IFNAR1 inhibits multiple myeloma–driven Treg expansion and immunosuppression
Yawara Kawano, … , Jamil Azzi, Irene M. Ghobrial
Yawara Kawano, … , Jamil Azzi, Irene M. Ghobrial
Published March 20, 2018
Citation Information: J Clin Invest. 2018;128(6):2487-2499. https://doi.org/10.1172/JCI88169.
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Research Article Hematology Immunology

Blocking IFNAR1 inhibits multiple myeloma–driven Treg expansion and immunosuppression

Abstract

Despite significant advances in the treatment of multiple myeloma (MM), most patients succumb to disease progression. One of the major immunosuppressive mechanisms that is believed to play a role in myeloma progression is the expansion of regulatory T cells (Tregs). In this study, we demonstrate that myeloma cells drive Treg expansion and activation by secreting type 1 interferon (IFN). Blocking IFN α and β receptor 1 (IFNAR1) on Tregs significantly decreases both myeloma-associated Treg immunosuppressive function and myeloma progression. Using syngeneic transplantable murine myeloma models and bone marrow (BM) aspirates of MM patients, we found that Tregs were expanded and activated in the BM microenvironment at early stages of myeloma development. Selective depletion of Tregs led to a complete remission and prolonged survival in mice injected with myeloma cells. Further analysis of the interaction between myeloma cells and Tregs using gene sequencing and enrichment analysis uncovered a feedback loop, wherein myeloma-cell-secreted type 1 IFN induced proliferation and expansion of Tregs. By using IFNAR1-blocking antibody treatment and IFNAR1-knockout Tregs, we demonstrated a significant decrease in myeloma-associated Treg proliferation, which was associated with longer survival of myeloma-injected mice. Our results thus suggest that blocking type 1 IFN signaling represents a potential strategy to target immunosuppressive Treg function in MM.

Authors

Yawara Kawano, Oksana Zavidij, Jihye Park, Michele Moschetta, Katsutoshi Kokubun, Tarek H. Mouhieddine, Salomon Manier, Yuji Mishima, Naoka Murakami, Mark Bustoros, Romanos Sklavenitis Pistofidis, Mairead Reidy, Yu J. Shen, Mahshid Rahmat, Pavlo Lukyanchykov, Esilida Sula Karreci, Shokichi Tsukamoto, Jiantao Shi, Satoshi Takagi, Daisy Huynh, Antonio Sacco, Yu-Tzu Tai, Marta Chesi, P. Leif Bergsagel, Aldo M. Roccaro, Jamil Azzi, Irene M. Ghobrial

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Figure 2

Upregulation of immune checkpoint molecules in Tregs in the BM of Vk*MYC-injected mice and SMM patients.

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Upregulation of immune checkpoint molecules in Tregs in the BM of Vk*MYC...
(A) BM Tregs of Vk*MYC-injected mice have significantly higher frequency of immune checkpoint receptor–expressing (PD1, LAG3, and TIM3) Tregs compared with control BM Tregs at the late time point (n = 3 per group, validated by FACS with n = 5 per group). (B) SPADE was conducted on BM Tregs of control and Vk*MYC-injected mice at the late time point. The nodes of the tree represent clusters of cells that are similar in marker expression. The color indicates the expression of LAG3. Increase of LAG3+ cells within Tregs was observed in Vk*MYC-injected BM. Heatmaps of immune checkpoint receptor (PD1, LAG3, and TIM3) expression of BM (C) and PB (D) Tregs at early and late time points. The color indicates the mean expression fold change of each molecule. BM Tregs of Vk*MYC-injected mice had higher expression of immune checkpoint receptors compared with control BM Tregs, while PB Tregs of Vk*MYC-injected mice did not show increased expression compared with control PB Tregs throughout disease progression. (F) Increased number of Tregs expressing immune checkpoint receptors in BM of SMM patients (n = 17) as compared with healthy donors (n = 11). Data combined from 3 independent experiments. NBM, normal bone marrow. (E) Heatmap of PD1, LAG3, and Tim3 expression on BM CD4+ and CD8+ T effector cells (Teffs, CD44++CD62Llo) at late time points. The color indicates the mean expression fold change of each molecule. BM Teffs of Vk*MYC-injected mice had higher expression of immune checkpoint receptors compared with control BM Teffs. (G) Heatmap of immune checkpoint molecules and CCR7-chemokine receptor expression of Tregs in BM of MM patients compared with healthy donors. The color indicates the ratio of mean expression of each molecule. Tregs of MM patients show elevated expression levels of immune checkpoint molecules. (H) CD4+ and CD8+ T effector cells in BM of MM patients have higher expression of immune checkpoint molecules compared with healthy donors. The color indicates the ratio of mean expression of each molecule. P values determined by 2-tailed t test. Error bars indicate SD.