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Hypoxia-induced tumor exosomes promote M2-like macrophage polarization of infiltrating myeloid cells and microRNA-mediated metabolic shift


Developing tumors rapidly outgrow their oxygen supply and are subject to hypoxia, which stimulates hypersecretion of tumor-derived exosomes that promote angiogenesis, metastasis, and immunosuppression, but the molecular mediators of these pathological effects remain poorly defined. Using quantitative proteomics, we identified that exosomes produced by hypoxic tumor cells are highly enriched in immunomodulatory proteins and chemokines including CSF-1, CCL2, FTH, FTL, and TGFβ. Modeling exosome effects on tumor-infiltrating immune cells, we observed a potent ability of these hypoxia-induced vesicles to influence macrophage recruitment and promote M2-like polarization both in vitro and in vivo. In addition, hypoxic, but not normoxic, tumor exosomes enhanced oxidative phosphorylation in bone marrow-derived macrophages via transfer of let-7a miRNA, resulting in suppression of the insulin-Akt-mTOR signaling pathway. Together, these data demonstrate that hypoxia promotes tumor secretion of biomolecule-loaded exosomes that can modify the immunometabolic profile of infiltrating monocyte-macrophages to better evade host immunity and enhance tumor progression.

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  1. 1.

    Park JE, Tan HS, Datta A, Lai RC, Zhang H, Meng W, et al. Hypoxic tumor cell modulates its microenvironment to enhance angiogenic and metastatic potential by secretion of proteins and exosomes. Mol Cell Proteom. 2010;9:1085–99.

    CAS  Article  Google Scholar 

  2. 2.

    Wilson WR, Hay MP. Targeting hypoxia in cancer therapy. Nat Rev Cancer. 2011;11:393–410.

    CAS  Article  Google Scholar 

  3. 3.

    Colombo M, Raposo G, Théry C. Biogenesis, secretion, and intercellular interactions of exosomes and other extracellular vesicles. Annu Rev Cell Dev Biol. 2014;30:255–89.

    CAS  Article  Google Scholar 

  4. 4.

    De Toro J, Herschlik L, Waldner C, Mongini C. Emerging roles of exosomes in normal and pathological conditions: new insights for diagnosis and therapeutic applications. Front Immunol. 2015;4:203.

    Google Scholar 

  5. 5.

    Simons M, Raposo G. Exosomes—vesicular carriers for intercellular communication. Curr Opin Cell Biol. 2009;21:575–81.

    CAS  Article  Google Scholar 

  6. 6.

    Liu Y, Gu Y, Han Y, Zhang Q, Jiang Z, Zhang X, et al. Tumor exosomal rnas promote lung pre-metastatic niche formation by activating alveolar epithelial TLR3 to recruit neutrophils. Cancer Cell. 2016;30:243–56.

    Article  Google Scholar 

  7. 7.

    Peinado H, Alečković M, Lavotshkin S, Matei I, Costa-Silva B, Moreno-Bueno G, et al. Melanoma exosomes educate bone marrow progenitor cells toward a pro-metastatic phenotype through MET. Nat Med. 2012;18:883–91.

    CAS  Article  Google Scholar 

  8. 8.

    Wu L, Zhang X, Zhang B, Shi H, Yuan X, Sun Y, et al. Exosomes derived from gastric cancer cells activate NF-κB pathway in macrophages to promote cancer progression. Tumor Biol. 2016;37:12169–80.

    CAS  Article  Google Scholar 

  9. 9.

    Boelens Mirjam C, Wu Tony J, Nabet Barzin Y, Xu B, Qiu Y, Yoon T, et al. Exosome transfer from stromal to breast cancer cells regulates therapy resistance pathways. Cell. 2014;159:499–513.

    CAS  Article  Google Scholar 

  10. 10.

    Hoffman RM. Stromal-cell and cancer-cell exosomes leading the metastatic exodus for the promised niche. Breast Cancer Res. 2013;15:310.

    Article  Google Scholar 

  11. 11.

    Robbins PD, Morelli AE. Regulation of immune responses by extracellular vesicles. Nat Rev Immunol. 2014;14:195–208.

    CAS  Article  Google Scholar 

  12. 12.

    Viaud S, Terme M, Flament C, Taieb J, André F, Novault S, et al. Dendritic cell-derived exosomes promote natural killer cell activation and proliferation: a role for NKG2D ligands and IL-15Rα. PLoS ONE. 2009;4:e4942.

    Article  Google Scholar 

  13. 13.

    Bretz NP, Ridinger J, Rupp A-K, Rimbach K, Keller S, Rupp C, et al. Body fluid exosomes promote secretion of inflammatory cytokines in monocytic cells via toll-like receptor signaling. J Biol Chem. 2013;288:36691–702.

    CAS  Article  Google Scholar 

  14. 14.

    Whiteside TL. Exosomes and tumor-mediated immune suppression. J Clin Invest. 2016;126:1216–23.

    Article  Google Scholar 

  15. 15.

    Espinoza JL, Takami A, Yoshioka K, Nakata K, Sato T, Kasahara Y, et al. Human microRNA-1245 down-regulates the NKG2D receptor in natural killer cells and impairs NKG2D-mediated functions. Haematologica. 2012;97:1295–303.

    CAS  Article  Google Scholar 

  16. 16.

    Zhou J, Wang S, Sun K, Chng W-J. The emerging roles of exosomes in leukemogeneis. Oncotarget. 2016;7:50698–707.

    PubMed  PubMed Central  Google Scholar 

  17. 17.

    Lobb RJ, Lima LG, Möller A. Exosomes: key mediators of metastasis and pre-metastatic niche formation. Semin Cell Dev Biol. 2017;67:3–10.

    CAS  Article  Google Scholar 

  18. 18.

    Kucharzewska P, Belting M. Emerging roles of extracellular vesicles in the adaptive response of tumour cells to microenvironmental stress. J Extracell Vesicles. 2013;2:1–10.

    Article  Google Scholar 

  19. 19.

    Villarroya-Beltri C, Baixauli F, Gutierrez-Vazquez C, Sanchez-Madrid F, Mittelbrunn M. Sorting it out: regulation of exosome loading. Semin Cancer Biol. 2014;28:3–13.

    CAS  Article  Google Scholar 

  20. 20.

    Wargo JA, Reddy SM, Reuben A, Sharma P. Monitoring immune responses in the tumor microenvironment. Curr Opin Immunol. 2016;41:23–31.

    CAS  Article  Google Scholar 

  21. 21.

    Storm MP, Sorrell I, Shipley R, Regan S, Luetchford KA, Sathish J, et al. Hollow fiber bioreactors for in vivo-like mammalian tissue culture. J Vis Exp. 2016:53431.

  22. 22.

    Thompson A, Schäfer J, Kuhn K, Kienle S, Schwarz J, Schmidt G, et al. Tandem mass tags: a novel quantification strategy for comparative analysis of complex protein mixtures by MS/MS. Anal Chem. 2003;75:1895–904.

    CAS  Article  Google Scholar 

  23. 23.

    Semenza GL. HIF-1 and tumor progression: pathophysiology and therapeutics. Trends Mol Med. 2002;8:S62–7.

    CAS  Article  Google Scholar 

  24. 24.

    Zhu Q, Wong AK, Krishnan A, Aure MR, Tadych A, Zhang R, et al. Targeted exploration and analysis of large cross-platform human transcriptomic compendia. Nat Methods. 2015;12:211–4.

    CAS  Article  Google Scholar 

  25. 25.

    Lin EY, Nguyen AV, Russell RG, Pollard JW. Colony-stimulating factor 1 promotes progression of mammary tumors to malignancy. J Exp Med. 2001;193:727–40.

    CAS  Article  Google Scholar 

  26. 26.

    Qian B-Z, Li J, Zhang H, Kitamura T, Zhang J, Campion LR, et al. CCL2 recruits inflammatory monocytes to facilitate breast tumor metastasis. Nature. 2011;475:222–5.

    CAS  Article  Google Scholar 

  27. 27.

    Gray CP, Arosio P, Hersey P. Association of increased levels of heavy-chain ferritin with increased CD4+CD25+regulatory T-cell levels in patients with melanoma. Clin Cancer Res. 2003;9:2551–9.

    CAS  PubMed  Google Scholar 

  28. 28.

    Marie JC, Letterio JJ, Gavin M, Rudensky AY. TGF-β1 maintains suppressor function and Foxp3 expression in CD4+CD25+regulatory T cells. J Exp Med. 2005;201:1061–7.

    CAS  Article  Google Scholar 

  29. 29.

    Kumar V, Patel S, Tcyganov E, Gabrilovich DI. The nature of myeloid-derived suppressor cells in the tumor microenvironment. Trends Immunol. 2016;37:208–20.

    CAS  Article  Google Scholar 

  30. 30.

    Ugel S, De Sanctis F, Mandruzzato S, Bronte V. Tumor-induced myeloid deviation: when myeloid-derived suppressor cells meet tumor-associated macrophages. J Clin Invest. 2015;125:3365–76.

    Article  Google Scholar 

  31. 31.

    Green CE, Liu T, Montel V, Hsiao G, Lester RD, Subramaniam S, et al. Chemoattractant signaling between tumor cells and macrophages regulates cancer cell migration, metastasis and neovascularization. PLoS ONE. 2009;4:e6713.

    Article  Google Scholar 

  32. 32.

    Damuzzo V, Pinton L, Desantis G, Solito S, Marigo I, Bronte V, et al. Complexity and challenges in defining myeloid-derived suppressor cells. Cytometry B Clin Cytom. 2015;88:77–91.

    CAS  Article  Google Scholar 

  33. 33.

    Kikushige Y, Yoshimoto G, Miyamoto T, Iino T, Mori Y, Iwasaki H, et al. Human Flt3 is expressed at the hematopoietic stem cell and the granulocyte/macrophage progenitor stages to maintain cell survival. J Immunol. 2008;180:7358–67.

    CAS  Article  Google Scholar 

  34. 34.

    Geeraerts X, Bolli E, Fendt S-M, Van Ginderachter JA. Macrophage metabolism as therapeutic target for cancer, atherosclerosis, and obesity. Front Immunol. 2017;8:289.

    Article  Google Scholar 

  35. 35.

    Mosser DM, Edwards JP. Exploring the full spectrum of macrophage activation. Nat Rev Immunol. 2008;8:958–69.

    CAS  Article  Google Scholar 

  36. 36.

    Roszer T. Understanding the mysterious M2 macrophage through activation markers and effector mechanisms. Mediators Inflamm. 2015;2015:816460.

    Article  Google Scholar 

  37. 37.

    Genin M, Clement F, Fattaccioli A, Raes M, Michiels C. M1 and M2 macrophages derived from THP-1 cells differentially modulate the response of cancer cells to etoposide. BMC Cancer. 2015;15:577–577.

    Article  Google Scholar 

  38. 38.

    Hu JM, Liu K, Liu JH, Jiang XL, Wang XL, Chen YZ, et al. CD163 as a marker of M2 macrophage, contribute to predicte aggressiveness and prognosis of Kazakh esophageal squamous cell carcinoma. Oncotarget. 2017;8:21526–38.

    PubMed  PubMed Central  Google Scholar 

  39. 39.

    Sudan B, Wacker MA, Wilson ME, Graff JW. A systematic approach to identify markers of distinctly activated human macrophages. Front Immunol. 2015;6:253.

  40. 40.

    Biswas Subhra K. Metabolic reprogramming of immune cells in cancer progression. Immunity. 2015;43:435–49.

    CAS  Article  Google Scholar 

  41. 41.

    Otero-Albiol D, Felipe-Abrio B. MicroRNA regulating metabolic reprogramming in tumor cells: new tumor markers. Cancer Transl Med. 2016;2:175–81.

    Article  Google Scholar 

  42. 42.

    Zhu H, Shyh-Chang N, Segrè Ayellet V, Shinoda G, Shah Samar P, Einhorn William S, et al. The Lin28/let-7 axis regulates glucose metabolism. Cell. 2011;147:81–94.

    CAS  Article  Google Scholar 

  43. 43.

    Lewis BP, Burge CB, Bartel DP. Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets. Cell. 2004;120:15–20.

    Article  Google Scholar 

  44. 44.

    Finger EC, Giaccia AJ. Hypoxia, inflammation, and the tumor microenvironment in metastatic disease. Cancer Metastasis Rev. 2010;29:285–93.

    CAS  Article  Google Scholar 

  45. 45.

    Kao J, Houck K, Fan Y, Haehnel I, Libutti SK, Kayton ML, et al. Characterization of a novel tumor-derived cytokine. Endothelial-monocyte activating polypeptide II. J Biol Chem. 1994;269:25106–19.

    CAS  PubMed  Google Scholar 

  46. 46.

    Kore RA, Edmondson JL, Jenkins SV, Jamshidi-Parsian A, Dings RPM, Reyna NS, et al. Hypoxia-derived exosomes induce putative altered pathways in biosynthesis and ion regulatory channels in glioblastoma cells. Biochem Biophys Rep. 2018;14:104–13.

    PubMed  PubMed Central  Google Scholar 

  47. 47.

    Elliott LA, Doherty GA, Sheahan K, Ryan EJ. Human tumor-infiltrating myeloid cells: phenotypic and functional diversity. Front Immunol. 2017;8:86.

    Article  Google Scholar 

  48. 48.

    Galván-Peña S, O’Neill LAJ. Metabolic reprograming in macrophage polarization. Front Immunol. 2014;5:420.

    PubMed  PubMed Central  Google Scholar 

  49. 49.

    O’Neill LAJ, Pearce EJ. Immunometabolism governs dendritic cell and macrophage function. J Exp Med. 2016;213:15–23.

    Article  Google Scholar 

  50. 50.

    Wenes M, Shang M, Di Matteo M, Goveia J, Martín-Pérez R, Serneels J, et al. Macrophage metabolism controls tumor blood vessel morphogenesis and metastasis. Cell Metab. 2016;24:701–15.

    CAS  Article  Google Scholar 

  51. 51.

    Jérôme T, Laurie P, Louis B, Pierre C. Enjoy the silence: the story of let-7 microRNA and cancer. Curr Genomics. 2007;8:229–33.

    Article  Google Scholar 

  52. 52.

    Young L, Sung J, Stacey G, Masters JR. Detection of mycoplasma in cell cultures. Nat Protoc. 2010;5:929.

    CAS  Article  Google Scholar 

  53. 53.

    Park JE, Sun Y, Lim SK, Tam JP, Dekker M, Chen H, et al. Dietary phytochemical PEITC restricts tumor development via modulation of epigenetic writers and erasers. Sci Rep. 2017;7:40569.

    CAS  Article  Google Scholar 

  54. 54.

    Hume DA, MacDonald KPA. Therapeutic applications of macrophage colony-stimulating factor-1 (CSF-1) and antagonists of CSF-1 receptor (CSF-1R) signaling. Blood. 2012;119:1810–20.

    CAS  Article  Google Scholar 

  55. 55.

    Murdoch C, Giannoudis A, Lewis CE. Mechanisms regulating the recruitment of macrophages into hypoxic areas of tumors and other ischemic tissues. Blood. 2004;104:2224–34.

    CAS  Article  Google Scholar 

  56. 56.

    Nielsen SR, Schmid MC. Macrophages as key drivers of cancer progression and metastasis. Mediators Inflamm. 2017;2017:11.

    Article  Google Scholar 

  57. 57.

    Li M, Knight DA, A Snyder L, Smyth MJ, Stewart TJ. A role for CCL2 in both tumor progression and immunosurveillance. Oncoimmunology. 2013;2:e25474.

    Article  Google Scholar 

  58. 58.

    Lee DD, Lal CV, Persad EA, Lowe C-W, Schwarz AM, Awasthi N, et al. Endothelial monocyte-activating polypeptide II mediates macrophage migration in the development of hyperoxia-induced lung disease of prematurity. Am J Respir Cell Mol Biol. 2016;55:602–12.

    CAS  Article  Google Scholar 

  59. 59.

    Jia W, Kidoya H, Yamakawa D, Naito H, Takakura N. Galectin-3 accelerates M2 macrophage infiltration and angiogenesis in tumors. Am J Pathol. 2013;182:1821–31.

    CAS  Article  Google Scholar 

  60. 60.

    Alkhateeb AA, Han B, Connor JR. Ferritin stimulates breast cancer cells through an iron-independent mechanism and is localized within tumor-associated macrophages. Breast Cancer Res Treat. 2013;137:733–44.

    CAS  Article  Google Scholar 

  61. 61.

    Gray CP, Arosio P, Hersey P. Heavy chain ferritin activates regulatory T cells by induction of changes in dendritic cells. Blood. 2002;99:3326–34.

    CAS  Article  Google Scholar 

  62. 62.

    Alkhateeb AA, Connor JR. The significance of ferritin in cancer: anti-oxidation, inflammation and tumorigenesis. Biochim Biophys Acta. 2013;1836:245–54.

    CAS  PubMed  Google Scholar 

  63. 63.

    Jezequel P, Campion L, Spyratos F, Loussouarn D, Campone M, Guerin-Charbonnel C, et al. Validation of tumor-associated macrophage ferritin light chain as a prognostic biomarker in node-negative breast cancer tumors: a multicentric 2004 national PHRC study. Int J Cancer. 2012;131:426–37.

    CAS  Article  Google Scholar 

  64. 64.

    Wu T, Li Y, Liu B, Zhang S, Wu L, Zhu X, et al. Expression of ferritin light chain (FTL) Is elevated in glioblastoma, and FTL silencing inhibits glioblastoma cell proliferation via the GADD45/JNK pathway. PLoS ONE. 2016;11:e0149361.

    Article  Google Scholar 

  65. 65.

    Bellomo C, Caja L, Moustakas A. Transforming growth factor β as regulator of cancer stemness and metastasis. Br J Cancer. 2016;115:761.

    CAS  Article  Google Scholar 

  66. 66.

    Lebrun J-J. The dual role of TGF in human cancer: from tumor suppression to cancer metastasis. ISRN Mol Biol. 2012;2012:28.

    Google Scholar 

  67. 67.

    Tauriello DVF, Palomo-Ponce S, Stork D, Berenguer-Llergo A, Badia-Ramentol J, Iglesias M, et al. TGFβ drives immune evasion in genetically reconstituted colon cancer metastasis. Nature. 2018;554:538.

    CAS  Article  Google Scholar 

  68. 68.

    Chen X, Wang S, Wu N, Yang CS. Leukotriene A4 hydrolase as a target for cancer prevention and therapy. Curr Cancer Drug Targets. 2004;4:267–83.

    CAS  Article  Google Scholar 

  69. 69.

    Vo TTL, Jang WJ, Jeong CH. Leukotriene A4 hydrolase: an emerging target of natural products for cancer chemoprevention and chemotherapy. Ann N Y Acad Sci. 2018;1431:3–13.

    Article  Google Scholar 

  70. 70.

    Balogh KN, Templeton DJ, Cross JV. Macrophage Migration Inhibitory Factor protects cancer cells from immunogenic cell death and impairs anti-tumor immune responses. PLoS ONE. 2018;13:e0197702.

    Article  Google Scholar 

  71. 71.

    Kindt N, Journe F, Laurent G, Saussez S. Involvement of macrophage migration inhibitory factor in cancer and novel therapeutic targets. Oncol Lett. 2016;12:2247–53.

    CAS  Article  Google Scholar 

  72. 72.

    Chen SC, Kung ML, Hu TH, Chen HY, Wu JC, Kuo HM, et al. Hepatoma-derived growth factor regulates breast cancer cell invasion by modulating epithelial--mesenchymal transition. J Pathol. 2012;228:158–69.

    CAS  Article  Google Scholar 

  73. 73.

    Gialeli C, Theocharis AD, Karamanos NK. Roles of matrix metalloproteinases in cancer progression and their pharmacological targeting. FEBS J. 2011;278:16–27.

    CAS  Article  Google Scholar 

  74. 74.

    Wang D, Zhang S, Chen F. High expression of PLOD1 drives tumorigenesis and affects clinical outcome in gastrointestinal carcinoma. Genet Test Mol Biomarkers. 2018;22:366–73.

    CAS  Article  Google Scholar 

  75. 75.

    Gilkes DM, Bajpai S, Wong CC, Chaturvedi P, Hubbi ME, Wirtz D, et al. Procollagen lysyl hydroxylase 2 is essential for hypoxia-induced breast cancer metastasis. Mol Cancer Res. 2013;11:456–66.

    CAS  Article  Google Scholar 

  76. 76.

    Mogami T, Yokota N, Asai-Sato M, Yamada R, Koizume S, Sakuma Y, et al. Annexin A4 is involved in proliferation, chemo-resistance and migration and invasion in ovarian clear cell adenocarcinoma cells. PLoS ONE. 2013;8:e80359.

    CAS  Article  Google Scholar 

  77. 77.

    Winter J, Diederichs S. Argonaute proteins regulate microRNA stability: Increased microRNA abundance by Argonaute proteins is due to microRNA stabilization. RNA Biol. 2011;8:1149–57.

    CAS  Article  Google Scholar 

  78. 78.

    Dueck A, Ziegler C, Eichner A, Berezikov E, Meister G. microRNAs associated with the different human Argonaute proteins. Nucleic Acids Res. 2012;40:9850–62.

    CAS  Article  Google Scholar 

  79. 79.

    Leca J, Martinez S, Lac S, Nigri J, Secq V, Rubis M, et al. Cancer-associated fibroblast-derived annexin A6+extracellular vesicles support pancreatic cancer aggressiveness. J Clin Invest. 2016;126:4140–56.

    Article  Google Scholar 

  80. 80.

    Qi H, Liu S, Guo C, Wang J, Greenaway FT, Sun M-Z. Role of annexin A6 in cancer. Oncol Lett. 2015;10:1947–52.

    CAS  Article  Google Scholar 

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This work is in part supported by grants from the Singapore Ministry of Education (MOE2014-T2-2-043, MOE2016-T2-2-018, and MOE2016-T3-1-003) and the National Medical Research Council of Singapore (NMRC-OF-IRG-0003-2016).

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JEP designed and performed the experiments, analyzed the data, and wrote the paper; BD performed the exosome TMT proteomics experiments and analyzed the data; SWT and NG performed animal experiments; CFT performed exosome preparation and analysis; and JKL, KWY, OLK, and JPT contributed to reagents and discussion; SSK conceived, designed, supervised the project, and revised the manuscript. All co-authors contributed to the revision of the manuscript.

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Correspondence to Jung Eun Park or Siu Kwan Sze.

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Park, J.E., Dutta, B., Tse, S.W. et al. Hypoxia-induced tumor exosomes promote M2-like macrophage polarization of infiltrating myeloid cells and microRNA-mediated metabolic shift. Oncogene 38, 5158–5173 (2019).

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