Abstract
Natural killer (NK) cells are commonly reduced in human tumors, enabling many to evade surveillance. Here, we sought to identify cues that alter NK cell activity in tumors. We found that, in human lung cancer, the presence of NK cells inversely correlated with that of monocyte-derived macrophages (mo-macs). In a murine model of lung adenocarcinoma, we show that engulfment of tumor debris by mo-macs triggers a pro-tumorigenic program governed by triggering receptor expressed on myeloid cells 2 (TREM2). Genetic deletion of Trem2 rescued NK cell accumulation and enabled an NK cell-mediated regression of lung tumors. TREM2+ mo-macs reduced NK cell activity by modulating interleukin (IL)-18/IL-18BP decoy interactions and IL-15 production. Notably, TREM2 blockade synergized with an NK cell-activating agent to further inhibit tumor growth. Altogether, our findings identify a new axis, in which TREM2+ mo-macs suppress NK cell accumulation and cytolytic activity. Dual targeting of macrophages and NK cells represents a new strategy to boost antitumor immunity.
This is a preview of subscription content, access via your institution
Access options
Access Nature and 54 other Nature Portfolio journals
Get Nature+, our best-value online-access subscription
$29.99 / 30 days
cancel any time
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
Data availability
All murine sequencing data will be made publicly available (GSE184304, GSE184309 and GSE184317). The human dataset is available at the Sequence Read Archive with BioProject accession no. PRJNA609924. Source data are provided with this paper.
References
Dunn, G. P., Bruce, A. T., Ikeda, H., Old, L. J. & Schreiber, R. D. Cancer immunoediting: from immunosurveillance to tumor escape. Nat. Immunol. 3, 991–998 (2002).
Molgora, M., Cortez, V. S. & Colonna, M. Killing the invaders: NK cell impact in tumors and anti-tumor therapy. Cancers 13, 595 (2021).
Gasser, S., Orsulic, S., Brown, E. J. & Raulet, D. H. The DNA damage pathway regulates innate immune system ligands of the NKG2D receptor. Nature 436, 1186–1190 (2005).
Raulet, D. H., Gasser, S., Gowen, B. G., Deng, W. & Jung, H. Regulation of ligands for the NKG2D activating receptor. Annu. Rev. Immunol. 31, 413–441 (2013).
Hayakawa, Y. & Smyth, M. J. NKG2D and cytotoxic effector function in tumor immune surveillance. Semin. Immunol. 18, 176–185 (2006).
Smyth, M. J., Hayakawa, Y., Takeda, K. & Yagita, H. New aspects of natural-killer-cell surveillance and therapy of cancer. Nat. Rev. Cancer 2, 850–861 (2002).
Parham, P. & Guethlein, L. A. Genetics of natural killer cells in human health, disease, and survival. Annu. Rev. Immunol. 36, 519–548 (2018).
Garrido, F., Aptsiauri, N., Doorduijn, E. M., Garcia Lora, A. M. & van Hall, T. The urgent need to recover MHC class I in cancers for effective immunotherapy. Curr. Opin. Immunol. 39, 44–51 (2016).
McGranahan, N. et al. Allele-specific HLA loss and immune escape in lung cancer evolution. Cell 171, 1259–1271 (2017).
Cózar, B. et al. Tumor-infiltrating natural killer cells. Cancer Discov. 11, 34–44 (2021).
Shimasaki, N., Jain, A. & Campana, D. NK cells for cancer immunotherapy. Nat. Rev. Drug Discov. 19, 200–218 (2020).
Lavin, Y. et al. Innate immune landscape in early lung adenocarcinoma by paired single-cell analyses. Cell 169, 750–765 (2017).
Leader, A. M. et al. Single-cell analysis of human non-small cell lung cancer lesions refines tumor classification and patient stratification. Cancer Cell 39, 1594–1609 (2021).
Mulder, K. et al. Cross-tissue single-cell landscape of human monocytes and macrophages in health and disease. Immunity 54, 1883–1900 (2021).
DeNardo, D. G. et al. CD4(+) T cells regulate pulmonary metastasis of mammary carcinomas by enhancing protumor properties of macrophages. Cancer Cell 16, 91–102 (2009).
Masetti, M. et al. Lipid-loaded tumor-associated macrophages sustain tumor growth and invasiveness in prostate cancer. J. Exp. Med. 219, e20210564 (2022).
Lavin, Y., Mortha, A., Rahman, A. & Merad, M. Regulation of macrophage development and function in peripheral tissues. Nat. Rev. Immunol. 15, 731–744 (2015).
Park, M. D., Silvin, A., Ginhoux, F. & Merad, M. Macrophages in health and disease. Cell 185, 4259–4279 (2022).
Casanova-Acebes, M. et al. Tissue-resident macrophages provide a pro-tumorigenic niche to early NSCLC cells. Nature 595, 578–584 (2021).
Loyher, P.-L. et al. Macrophages of distinct origins contribute to tumor development in the lung. J. Exp. Med. 215, 2536–2553 (2018).
Guerreiro, R. et al. TREM2 variants in Alzheimer’s disease. N. Engl. J. Med. 368, 117–127 (2013).
Sims, R. et al. Rare coding variants in PLCG2, ABI3, and TREM2 implicate microglial-mediated innate immunity in Alzheimer’s disease. Nat. Genet. 49, 1373–1384 (2017).
Wang, S. et al. Anti-human TREM2 induces microglia proliferation and reduces pathology in an Alzheimer’s disease model. J. Exp. Med. 217, e20200785 (2020).
Zhou, Y. et al. Human and mouse single-nucleus transcriptomics reveal TREM2-dependent and TREM2-independent cellular responses in Alzheimer’s disease. Nat. Med. 26, 131–142 (2020).
Binnewies, M. et al. Targeting TREM2 on tumor-associated macrophages enhances immunotherapy. Cell Rep. 37, 109844 (2021).
Katzenelenbogen, Y. et al. Coupled scRNA-seq and Intracellular protein activity reveal an immunosuppressive role of TREM2 in cancer. Cell 182, 872–885 (2020).
Molgora, M. et al. TREM2 modulation remodels the tumor myeloid landscape enhancing anti-PD-1 immunotherapy. Cell 182, 886–900 (2020).
Esparza-Baquer, A. et al. TREM-2 defends the liver against hepatocellular carcinoma through multifactorial protective mechanisms. Gut 70, 1345–1361 (2021).
Perugorria, M. J. et al. Non-parenchymal TREM-2 protects the liver from immune-mediated hepatocellular damage. Gut 68, 533–546 (2019).
Tang, W. et al. TREM2 acts as a tumor suppressor in hepatocellular carcinoma by targeting the PI3K/Akt/β-catenin pathway. Oncogenesis 8, 9 (2019).
DuPage, M., Dooley, A. L. & Jacks, T. Conditional mouse lung cancer models using adenoviral or lentiviral delivery of Cre recombinase. Nat. Protoc. 4, 1064–1072 (2009).
Heng, T. S. P. & Painter, M. W. The Immunological Genome Project: networks of gene expression in immune cells. Nat. Immunol. 9, 1091–1094 (2008).
Cannon, J. P., O’Driscoll, M. & Litman, G. W. Specific lipid recognition is a general feature of CD300 and TREM molecules. Immunogenetics 64, 39–47 (2012).
Daws, M. R. et al. Pattern recognition by TREM-2: binding of anionic ligands. J. Immunol. 171, 594–599 (2003).
Filipello, F. et al. The microglial innate immune receptor TREM2 is required for synapse elimination and normal brain connectivity. Immunity 48, 979–991 (2018).
Hsieh, C. L. et al. A role for TREM2 ligands in the phagocytosis of apoptotic neuronal cells by microglia. J. Neurochem. 109, 1144–1156 (2009).
Iizasa E. et al. TREM2 is a receptor for non-glycosylated mycolic acids of mycobacteria that limits anti-mycobacterial macrophage activation. Nat. Commun. 12, 2299 (2021).
Lue, L.-F., Schmitz, C. & Walker, D. G. What happens to microglial TREM2 in Alzheimer’s disease: Immunoregulatory turned into immunopathogenic? Neuroscience 302, 138–150 (2015).
Shirotani, K. et al. Aminophospholipids are signal-transducing TREM2 ligands on apoptotic cells. Sci. Rep. 9, 7508 (2019).
Takahashi, K., Rochford, C. D. P. & Neumann, H. Clearance of apoptotic neurons without inflammation by microglial triggering receptor expressed on myeloid cells-2. J. Exp. Med. 201, 647–657 (2005).
Zhang, S. et al. Efferocytosis fuels requirements of fatty acid oxidation and the electron transport chain to polarize macrophages for tissue repair. Cell Metab. 29, 443–456 (2019).
Hannedouche, S. et al. Oxysterols direct immune cell migration via EBI2. Nature 475, 524–527 (2011).
Preuss, I. et al. Transcriptional regulation and functional characterization of the oxysterol/EBI2 system in primary human macrophages. Biochem. Biophys. Res. Commun. 446, 663–668 (2014).
Rutkowska, A. et al. The EBI2 signalling pathway plays a role in cellular crosstalk between astrocytes and macrophages. Sci. Rep. 6, 25520 (2016).
Harms, R. Z., Creer, A. J., Lorenzo-Arteaga, K. M., Ostlund, K. R. & Sarvetnick, N. E. Interleukin (IL)-18 binding protein deficiency disrupts natural killer cell maturation and diminishes circulating IL-18. Front. Immunol. 8, 1020 (2017).
Zhou, T. et al. IL-18BP is a secreted immune checkpoint and barrier to IL-18 immunotherapy. Nature 583, 609–614 (2020).
Chaix, J. et al. Cutting edge: priming of NK cells by IL-18. J. Immunol. 181, 1627–1631 (2008).
Nielsen, C. M., Wolf, A.-S., Goodier, M. R. & Riley, E. M. Synergy between common γ chain family cytokines and IL-18 potentiates innate and adaptive pathways of NK cell activation. Front. Immunol. 7, 101 (2016).
Rautela, J. & Huntington, N. D. IL-15 signaling in NK cell cancer immunotherapy. Curr. Opin. Immunol. 44, 1–6 (2017).
Ferrari de Andrade, L. et al. Antibody-mediated inhibition of MICA and MICB shedding promotes NK cell-driven tumor immunity. Science 359, 1537–1542 (2018).
Izawa, S. et al. H2O2 production within tumor microenvironment inversely correlated with infiltration of CD56(dim) NK cells in gastric and esophageal cancer: possible mechanisms of NK cell dysfunction. Cancer Immunol. Immunother. 60, 1801–1810 (2011).
Jin, S. et al. NK cell phenotypic modulation in lung cancer environment. PLoS ONE 9, e109976 (2014).
Sconocchia, G. et al. NK cells and T cells cooperate during the clinical course of colorectal cancer. Oncoimmunology 3, e952197 (2014).
Stankovic, B. et al. Immune cell composition in human non-small cell lung cancer. Front. Immunol. 9, 3101 (2018).
Bonacina, F. et al. Myeloid apolipoprotein E controls dendritic cell antigen presentation and T cell activation. Nat. Commun. 9, 3083 (2018).
Gouna, G. et al. TREM2-dependent lipid droplet biogenesis in phagocytes is required for remyelination. J. Exp. Med. 218, e20210227 (2021).
van Eijk, M. & Aerts, J. M. F. G. The unique phenotype of lipid-laden macrophages. Int. J. Mol. Sci. 22, 4039 (2021).
Di Conza, G. et al. Tumor-induced reshuffling of lipid composition on the endoplasmic reticulum membrane sustains macrophage survival and pro-tumorigenic activity. Nat. Immunol. 22, 1403–1415 (2021).
Nugent, A. A. et al. TREM2 regulates microglial cholesterol metabolism upon chronic phagocytic challenge. Neuron 105, 837–854 (2020).
Jordan, J. A. et al. Role of IL-18 in acute lung inflammation. J. Immunol. 167, 7060–7068 (2001).
Pechkovsky, D. V., Goldmann, T., Vollmer, E., Müller-Quernheim, J. & Zissel, G. Interleukin-18 expression by alveolar epithelial cells type II in tuberculosis and sarcoidosis. FEMS Immunol. Med. Microbiol. 46, 30–38 (2006).
Barry, K. C. et al. A natural killer–dendritic cell axis defines checkpoint therapy–responsive tumor microenvironments. Nat. Med. 24, 1178–1191 (2018).
Böttcher, J. P. et al. NK cells stimulate recruitment of cDC1 into the tumor microenvironment promoting cancer immune control. Cell 172, 1022–1037 (2018).
Mattiuz, R. et al. Type 1 conventional dendritic cells and interferons are required for spontaneous CD4+ and CD8+ T-cell protective responses to breast cancer. Clin. Transl. Immunol. 10, e1305 (2021).
Bonnardel, J. et al. Stellate cells, hepatocytes, and endothelial cells imprint the Kupffer cell identity on monocytes colonizing the liver macrophage niche. Immunity 51, 638–654 (2019).
Browaeys, R., Saelens, W. & Saeys, Y. NicheNet: modeling intercellular communication by linking ligands to target genes. Nat. Methods 17, 159–162 (2019).
Acknowledgements
We thank members of the Merad and Brown laboratories at the Marc and Jennifer Lipschultz Precision Immunology Institute at Mount Sinai and the Tisch Cancer Institute for insightful discussions and feedback; we specifically thank M.J. Lin and J. Brody for lending us flow cytometry antibodies against the NKG2D ligands; the Mount Sinai Flow Cytometry Core, the Human Immune Monitoring Center and the Mount Sinai Biorepository for support. Data in this paper were used in a dissertation as partial fulfillment of the requirements for a PhD at the Graduate School of Biomedical Sciences at Mount Sinai.
Author information
Authors and Affiliations
Contributions
M.M. conceived the project. M.M., M.D.P. and I.R.T. wrote the manuscript. I.R.T., M.D.P. and M.M. designed the experiments. I.R.T., M.D.P., J.L.B., N.M.L., E.H., S.H., M.B., A.N. and A.R.S. performed experiments. I.R.T., M.D.P., J.L.B. and A.N. maintained mouse colonies and cell cultures for these experiments. M.D.P. and A.M. performed computational analyses, with additional support from T.D., D.D. and S.H. P.H., L.T. and J.G. provided immunohistochemistry stains and analyses. M.M., J.H. and M.C. provided the TREM2 blocking antibody and isotype control. J.N., A.F. and N.B. provided the IL-15 neutralizing antibody; M.J.L. and J.B. provided the antibodies against NKG2D ligands; and L.F.A. provided the MIC-A stabilizing antibody and B16-F10-MICA cell line. A.R.S., B.M., J.C.M., E.K., A.O.K., M.C.A., N.B., A.H., L.F.A., B.D.B., M.C. and T.U.M. provided intellectual input. This work was supported by National Institutes of Health grants R01 AI153363 (to A.O.K.), R01 CA254104 and R01 CA257195 (to M.M. and B.D.B.), 1R37 CA269982-01A1 (to L.F.d.A.), R01CA262684 to M.C. S.H. was supported by the National Cancer Institute predoctoral-to-postdoctoral fellowship K00 CA223043. N.M.L. was supported by the Cancer Research Institute/Bristol Myers Squibb Irvington Postdoctoral Research Fellowship to Promote Racial Diversity (award no. CRI3931). B.D.B. and M.M. were supported by the Applebaum Foundation and the Feldman Foundation. B.D.B. was supported by the Alliance for Cancer Gene Therapy. L.F.d.A. is the recipient of a Cancer Research Institute Clinic and Laboratory Integration Program Grant (award no. CRI3483), Tisch Cancer Institute Developments Funds Award (P30CA196521), Department of Defense Career Development Award (W81XWH2210262 and project number CA210940), Leukemia and Lymphoma Society (award no. 6647-23) and is supported by the Elsa U. Pardee Foundation.
Corresponding author
Ethics declarations
Competing interests
L.F.d.A. is a named inventor of the MIC-A therapeutic antibody in a filed patient (US20200165343A1) and has received royalty payments in relation to this therapeutic. L.F.d.A. is also co-inventor in issued patents about an α-3 domain-specific antibody. M.C. receives research support from NGM Biotechnology and Vigil Neuro, is a scientific advisory board member of NGM Biotechnology and Vigil Neuro and has a patent for TREM2 pending. M.M. serves on the scientific advisory board and holds stock from Compugen, Myeloid Therapeutics, Morphic Therapeutic, Asher Bio, Dren Bio, Nirogy, Oncoresponse, Owkin, Pionyr, OSE and Larkspur. M.M. serves on the scientific advisory board Innate Pharma, DBV and Genenta. M.M. is a named co-inventor on an issued patent for multiplex immunohistochemistry to characterize tumors and treatment responses. The technology is filed through Icahn School of Medicine at Mount Sinai and is currently unlicensed. This technology was used to evaluate tissue in this study and the results could impact the value of this technology. The remaining authors declare no competing interests.
Peer review
Peer review information
Nature Immunology thanks Florian Greten and the other, anonymous, reviewer(s) for their contribution to the peer review of this work. Primary Handling Editor: S. Houston in collaboration with the Nature Immunology team.
Additional information
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Extended data
Extended Data Fig. 1 Purification and active signaling pathways in GFPlo and GFPhi mo-macs.
(a) Flow cytometry sort strategy (left) for purifying GFPhi and GFPlo mo-macs from single-cell suspensions of digested KP-GFP tumor-bearing lungs at four weeks post-inoculation of KP-GFP cells. Histograms show the fluorescence spectrum of GFP in mo-macs from tumor-bearing lungs, comprised of GFPhi (green) and GFPlo (blue) mo-macs. Each row represents an individual replicate. Representative confocal immunofluorescence (IF) imaging of FACS-sorted GFPlo and GFPhi mo-macs (right). For each pair of images, a broad-field view with a region of interest (ROI, white outline) (left) and a magnification of the ROI (right) are shown. Data is representative of two independent experiments. (b) Gene set enrichment analysis (GSEA) (Broad Institute) for pathways annotated in the KEGG and Reactome databases performed on the significantly differentially regulated genes in GFPhi mo-macs, relative to GFPlo mo-macs. The top 15 most significantly enriched terms are shown, with notable pathways highlighted in red for up-regulated genes in GFPhi mo-macs (left) and up-regulated genes in GFPlo mo-macs (right).
Extended Data Fig. 2 scRNA-seq profiling of mo-macs in chimeric mice.
(a) Expression of Trem2 and Gpnmb, as hallmark genes of the TREM2 gene program, across cell subtypes (mo-macs, AMs, Ly6Clo and Ly6Chi monocytes, mregDCs, cDC1 and cDC2) identified by unsupervised clustering. (b) Flow cytometry sort strategy to purify CD45.1 WT and CD45.2 KO fractions of MNPs from KP-GFP tumor-bearing chimeric mice. (c) Flow cytometric quantification of GFP+ mo-macs, represented as relative frequency among total mo-macs from the purified CD45.1 WT and CD45.2 KO fractions of MNPs from KP-GFP tumor-bearing mice. Paired values shown for paired CD45.1 WT and CD45.2 KO cells that were purified from the same biological replicate. (mean ± standard error of mean (S.E.M.); paired two-tailed t-test at a 95% confidence interval).
Extended Data Fig. 3 Contributions of CD8 T cells and NK cells to tumor regression in KO mice.
(a) Flow cytometric quantification of IFN-ɣ/TNF-α-producing CD8 T cells, as a relative frequency of total antigen-specific CD8 T cells, in the tumor-bearing lungs (left) and in the tumor-draining lymph nodes (tdLN) (right) of KP-GFP tumor-bearing WT and KO mice. (n = 4-5 mice per group, mean ± standard error of mean (S.E.M.); unpaired two-tailed t-test at a 95% confidence interval). (b) Flow cytometric quantification of NK cells expressing activation/degranulation markers, as a relative frequency of total NK cells. (n = 4-5 mice per group, mean ± standard error of mean (S.E.M.); unpaired two-tailed t-test at a 95% confidence interval). (c) Flow cytometric quantification of type 1 conventional dendritic cells (cDC1), as a relative frequency of CD45+ immune cells, in the tumor-bearing lungs of WT mice and KO mice treated with either an isotype control or CD8 (right) (n = 4-5 mice per group) or NK cell (left)-depleting antibody (n = 8-10 mice per group). (mean ± standard error of mean (S.E.M.); unpaired two-tailed t-test at a 95% confidence interval. Data is representative of two independent experiments.). (d) Quantification of the tumor area as a percent of the total area of the lung cross-section from tumor-bearing lungs of WT and KO mice treated with either an isotype control of NK cell-depleting antibody. (n = 8-10 mice per group, mean ± standard error of mean (S.E.M.); unpaired two-tailed t-test at a 95% confidence interval. Data shown is of at least two independent experiments.). (e) Flow cytometric quantification of NK cells, as a relative frequency of CD45+ immune cells. (n = 8-10 mice per group, mean ± standard error of mean (S.E.M.); unpaired two-tailed t-test at a 95% confidence interval. Data is representative of at least three independent experiments.). (f) Flow cytometric quantification of IFN-ɣ-producing (left) and TNF-α-producing (right) CD8 T cells in the tumor-bearing lungs of WT mice with isotype antibody, KO mice with isotype antibody, and KO mice given IL-18 neutralizing antibody. Data shown as a relative frequency among total CD8 T cells. (n = 5-8 mice per group, mean ± standard error of mean (S.E.M.); unpaired two-tailed t-test at a 95% confidence interval. Data shown is representative of at least three independent experiments).
Extended Data Fig. 4 Cooperative IL-15 signaling is needed to confer the therapeutic benefit of a TREM2-deficient setting.
(a) Relative expression of Il15 by mature mregDCs, as determined by scRNA-seq profiling of MNPs in the tumor-bearing lungs of WT and KO mice (Fig. 3C). (b) Representative H&E images of tumor-bearing lungs of WT, isotype antibody (n = 4), WT mice that received the IL-15 neutralizing antibody (n = 4), KO, isotype antibody (n = 4), KO mice that received the IL-15 neutralizing antibody (n = 5) (left). Quantification of the tumor area as a percent of the total area of the lung cross-section (right) is shown. (mean ± standard error of mean (S.E.M.); unpaired two-tailed t-test at a 95% confidence interval). (c) Flow cytometric quantification of NK cells, as a relative frequency of CD45+ immune cells, in the KP-GFP tumor-bearing lungs of isotype control WT and KO mice and WT and KO mice treated with the αIL-15 antibody. (n = 4-5 mice per group, mean ± standard error of mean (S.E.M.); unpaired two-tailed t-test at a 95% confidence interval. Data shown is representative of two independent experiments).
Supplementary information
Supplementary Tables
Supplementary Table 1. Differentially expressed genes between tissue-resident alveolar macrophages and monocyte-derived macrophages in NSCLC. Supplementary Table 2. Gene expression across dendritic cell, monocyte, macrophage and lymphoid cell subsets in NSCLC. Supplementary Table 3. Differentially expressed genes between CD45.1 versus CD45.2 monocyte-derived macrophages identified by scRNA-seq of chimeric mice (day 28 after inoculation of tumor cells).
Source data
Source Data Fig. 1
Statistical source data.
Source Data Fig. 2
Statistical source data.
Source Data Fig. 3
Statistical source data.
Source Data Fig. 4
Statistical source data.
Source Data Fig. 5
Statistical source data.
Source Data Extended Data Fig. 2
Statistical source data.
Source Data Extended Data Fig. 3
Statistical source data.
Source Data Extended Data Fig. 4
Statistical source data.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Park, M.D., Reyes-Torres, I., LeBerichel, J. et al. TREM2 macrophages drive NK cell paucity and dysfunction in lung cancer. Nat Immunol 24, 792–801 (2023). https://doi.org/10.1038/s41590-023-01475-4
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/s41590-023-01475-4
This article is cited by
-
Neutral ceramidase regulates breast cancer progression by metabolic programming of TREM2-associated macrophages
Nature Communications (2024)
-
An IL-4 signalling axis in bone marrow drives pro-tumorigenic myelopoiesis
Nature (2024)
-
Supervised discovery of interpretable gene programs from single-cell data
Nature Biotechnology (2023)