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The miR-126–VEGFR2 axis controls the innate response to pathogen-associated nucleic acids

Nature Immunology volume 15, pages 54–62 (2014)Cite this article

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Abstract

miR-126 is a microRNA expressed predominately by endothelial cells and controls angiogenesis. We found miR-126 was required for the innate response to pathogen-associated nucleic acids and that miR-126-deficient mice had greater susceptibility to infection with pseudotyped HIV. Profiling of miRNA indicated that miR-126 had high and specific expression by plasmacytoid dendritic cells (pDCs). Moreover, miR-126 controlled the survival and function of pDCs and regulated the expression of genes encoding molecules involved in the innate response, including Tlr7, Tlr9 and Nfkb1, as well as Kdr, which encodes the growth factor receptor VEGFR2. Deletion of Kdr in DCs resulted in reduced production of type I interferon, which supports the proposal of a role for VEGFR2 in miR-126 regulation of pDCs. Our studies identify the miR-126–VEGFR2 axis as an important regulator of the innate response that operates through multiscale control of pDCs.

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Figure 1: The IFN-α/β response is impaired in mice deficient in miR-126.
Figure 2: High and specific expression of miR-126 by pDCs.
Figure 3: Loss of miR-126 impairs the homeostasis of pDCs.
Figure 4: miR-126 controls the survival of pDCs.
Figure 5: Functional impairment of pDCs in the absence of miR-126.
Figure 6: miR-126 directly targets the mTOR pathway and indirectly regulates genes encoding key molecules of the innate response in pDCs.
Figure 7: VEGFR2 is important for pDC function and its expression is regulated by miR-126.

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References

  1. McCartney, S.A. & Colonna, M. Viral sensors: diversity in pathogen recognition. Immunol. Rev. 227, 87–94 (2009).

    Article  CAS  PubMed  Google Scholar 

  2. Kawai, T. & Akira, S. Toll-like receptors and their crosstalk with other innate receptors in infection and immunity. Immunity 34, 637–650 (2011).

    Article  CAS  PubMed  Google Scholar 

  3. Chevrier, N. et al. Systematic discovery of TLR signaling components delineates viral-sensing circuits. Cell 147, 853–867 (2011).

    Article  CAS  PubMed  Google Scholar 

  4. Yang, K. et al. Human TLR-7-, -8-, and -9-mediated induction of IFN-α/β and -λ is IRAK-4 dependent and redundant for protective immunity to viruses. Immunity 23, 465–478 (2005).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Kumar, H., Kawai, T. & Akira, S. Toll-like receptors and innate immunity. Biochem. Biophys. Res. Commun. 388, 621–625 (2009).

    Article  CAS  PubMed  Google Scholar 

  6. Baltimore, D. et al. MicroRNAs: new regulators of immune cell development and function. Nat. Immunol. 9, 839–845 (2008).

    Article  CAS  PubMed  Google Scholar 

  7. Bartel, D.P. MicroRNAs: target recognition and regulatory functions. Cell 136, 215–233 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Taganov, K.D., Boldin, M.P., Chang, K.-J. & Baltimore, D. NF-κB-dependent induction of microRNA miR-146, an inhibitor targeted to signaling proteins of innate immune responses. Proc. Natl. Acad. Sci. USA 103, 12481–12486 (2006).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. O'Connell, R.M. et al. MicroRNA-155 is induced during the macrophage inflammatory response. Proc. Natl. Acad. Sci. USA 104, 1604–1609 (2007).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Brown, B.D. et al. Endogenous microRNA can be broadly exploited to regulate transgene expression according to tissue, lineage and differentiation state. Nat. Biotechnol. 25, 1457–1467 (2007).

    Article  CAS  PubMed  Google Scholar 

  11. O'Connell, R.M., Chaudhuri, A.A., Rao, D.S. & Baltimore, D. Inositol phosphatase SHIP1 is a primary target of miR-155. Proc. Natl. Acad. Sci. USA 106, 7113–7118 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Zhou, H. et al. miR-155 and its star-form partner miR-155* cooperatively regulate type I interferon production by human plasmacytoid dendritic cells. Blood 116, 5885–5894 (2010).

    Article  CAS  PubMed  Google Scholar 

  13. O'Neill, L.A., Sheedy, F.J. & McCoy, C.E. MicroRNAs: the fine-tuners of Toll-like receptor signalling. Nat. Rev. Immunol. 11, 163–175 (2011).

    Article  CAS  PubMed  Google Scholar 

  14. Fish, J.E. et al. miR-126 regulates angiogenic signaling and vascular integrity. Dev. Cell 15, 272–284 (2008).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Kuhnert, F. et al. Attribution of vascular phenotypes of the murine Egfl7 locus to the microRNA miR-126. Development 135, 3989–3993 (2008).

    Article  CAS  PubMed  Google Scholar 

  16. Wang, S. et al. The endothelial-specific microRNA miR-126 governs vascular integrity and angiogenesis. Dev. Cell 15, 261–271 (2008).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  17. Brown, B.D. et al. In vivo administration of lentiviral vectors triggers a type I interferon response that restricts hepatocyte gene transfer and promotes vector clearance. Blood 109, 2797–2805 (2007).

    Article  CAS  PubMed  Google Scholar 

  18. Beignon, A.-S. et al. Endocytosis of HIV-1 activates plasmacytoid dendritic cells via Toll-like receptor-viral RNA interactions. J. Clin. Invest. 115, 3265–3275 (2005).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Meylan, P.R., Guatelli, J.C., Munis, J.R., Richman, D.D. & Kornbluth, R.S. Mechanisms for the inhibition of HIV replication by interferons-α, -β, and -γ in primary human macrophages. Virology 193, 138–148 (1993).

    Article  CAS  PubMed  Google Scholar 

  20. Reizis, B., Bunin, A., Ghosh, H.S., Lewis, K.L. & Sisirak, V. Plasmacytoid dendritic cells: recent progress and open questions. Annu. Rev. Immunol. 29, 163–183 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Colonna, M., Trinchieri, G. & Liu, Y.-J. Plasmacytoid dendritic cells in immunity. Nat. Immunol. 5, 1219–1226 (2004).

    Article  CAS  PubMed  Google Scholar 

  22. Naik, S.H. et al. Development of plasmacytoid and conventional dendritic cell subtypes from single precursor cells derived in vitro and in vivo. Nat. Immunol. 8, 1217–1226 (2007).

    Article  CAS  PubMed  Google Scholar 

  23. Cella, M. et al. Plasmacytoid monocytes migrate to inflamed lymph nodes and produce large amounts of type I interferon. Nat. Med. 5, 919–923 (1999).

    Article  CAS  PubMed  Google Scholar 

  24. Asselin-Paturel, C. et al. Type I interferon dependence of plasmacytoid dendritic cell activation and migration. J. Exp. Med. 201, 1157–1167 (2005).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Vitour, D. & Meurs, E.F. Regulation of interferon production by RIG-I and LGP2: a lesson in self-control. Sci. STKE 2007, pe20 (2007).

    Article  PubMed  Google Scholar 

  26. Miller, J.C. et al. Deciphering the transcriptional network of the dendritic cell lineage. Nat. Immunol. 13, 888–899 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Krug, A. et al. IFN-producing cells respond to CXCR3 ligands in the presence of CXCL12 and secrete inflammatory chemokines upon activation. J. Immunol. 169, 6079–6083 (2002).

    Article  CAS  PubMed  Google Scholar 

  28. Li, H.S. et al. The signal transducers STAT5 and STAT3 control expression of Id2 and E2–2 during dendritic cell development. Blood 120, 4363–4373 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. van de Laar, L. et al. PI3K-PKB hyperactivation augments human plasmacytoid dendritic cell development and function. Blood 120, 4982–4991 (2012).

    Article  CAS  PubMed  Google Scholar 

  30. Cao, W. et al. Toll-like receptor-mediated induction of type I interferon in plasmacytoid dendritic cells requires the rapamycin-sensitive PI(3)K-mTOR-p70S6K pathway. Nat. Immunol. 9, 1157–1164 (2008).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Quinn, T.P., Peters, K.G., De Vries, C., Ferrara, N. & Williams, L.T. Fetal liver kinase 1 is a receptor for vascular endothelial growth factor and is selectively expressed in vascular endothelium. Proc. Natl. Acad. Sci. USA 90, 7533–7537 (1993).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Carmeliet, P. & Jain, R.K. Molecular mechanisms and clinical applications of angiogenesis. Nature 473, 298–307 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Ding, B.-S. et al. Inductive angiocrine signals from sinusoidal endothelium are required for liver regeneration. Nature 468, 310–315 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Ghosh, H.S., Cisse, B., Bunin, A., Lewis, K.L. & Reizis, B. Continuous expression of the transcription factor e2–2 maintains the cell fate of mature plasmacytoid dendritic cells. Immunity 33, 905–916 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Grouard, G. et al. The enigmatic plasmacytoid T cells develop into dendritic cells with interleukin (IL)-3 and CD40-ligand. J. Exp. Med. 185, 1101–1111 (1997).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Siegal, F.P. et al. The nature of the principal type 1 interferon-producing cells in human blood. Science 284, 1835–1837 (1999).

    Article  CAS  PubMed  Google Scholar 

  37. Stepkowski, S.M., Chen, W., Ross, J.A., Nagy, Z.S. & Kirken, R.A. STAT3: an important regulator of multiple cytokine functions. Transplantation 85, 1372–1377 (2008).

    Article  CAS  PubMed  Google Scholar 

  38. Plas, D.R. & Thomas, G. Tubers and tumors: rapamycin therapy for benign and malignant tumors. Curr. Opin. Cell Biol. 21, 230–236 (2009).

    Article  CAS  PubMed  Google Scholar 

  39. Guiducci, C. et al. PI3K is critical for the nuclear translocation of IRF-7 and type I IFN production by human plasmacytoid predendritic cells in response to TLR activation. J. Exp. Med. 205, 315–322 (2008).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Conrad, C. et al. Plasmacytoid dendritic cells promote immunosuppression in ovarian cancer via ICOS costimulation of Foxp3+ T-regulatory cells. Cancer Res. 72, 5240–5249 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Ferrara, N., Mass, R.D., Campa, C. & Kim, R. Targeting VEGF-A to treat cancer and age-related macular degeneration. Annu. Rev. Med. 58, 491–504 (2007).

    Article  CAS  PubMed  Google Scholar 

  42. Nestle, F.O. et al. Plasmacytoid predendritic cells initiate psoriasis through interferon-alpha production. J. Exp. Med. 202, 135–143 (2005).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Lande, R. et al. Plasmacytoid dendritic cells sense self-DNA coupled with antimicrobial peptide. Nature 449, 564–569 (2007).

    Article  CAS  PubMed  Google Scholar 

  44. Guiducci, C. et al. Autoimmune skin inflammation is dependent on plasmacytoid dendritic cell activation by nucleic acids via TLR7 and TLR9. J. Exp. Med. 207, 2931–2942 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Diana, J. et al. Crosstalk between neutrophils, B-1a cells and plasmacytoid dendritic cells initiates autoimmune diabetes. Nat. Med. 19, 65–73 (2013).

    Article  CAS  PubMed  Google Scholar 

  46. Banchereau, J. & Pascual, V. Type I interferon in systemic lupus erythematosus and other autoimmune diseases. Immunity 25, 383–392 (2006).

    Article  CAS  PubMed  Google Scholar 

  47. Wang, H., Peng, W., Ouyang, X., Li, W. & Dai, Y. Circulating microRNAs as candidate biomarkers in patients with systemic lupus erythematosus. Transl. Res. 160, 198–206 (2012).

    Article  CAS  PubMed  Google Scholar 

  48. Janssen, H.L.A. et al. Treatment of HCV infection by targeting microRNA. N. Engl. J. Med. 368, 1685–1694 (2013).

    Article  CAS  PubMed  Google Scholar 

  49. O'Connell, R.M. et al. MicroRNA-155 promotes autoimmune inflammation by enhancing inflammatory T cell development. Immunity 33, 607–619 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. Agudo, J. et al. A TLR and non-TLR mediated innate response to lentiviruses restricts hepatocyte entry and can be ameliorated by pharmacological blockade. Mol. Ther. 20, 2257–2267 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

We thank C.J. Kuo (Stanford University) for Mir126−/− mice; P. Sathe, R. Sachidanandam, L. Naldini and B. Gentner for discussions; J. Ochando for reading the manuscript; and the Mount Sinai Mouse Genetics and Mouse Targeting facility, the Flow Cytometry Core and the Mount Sinai Genomics Core for technical assistance. Supported by the US National Institutes of Health (DP2DK083052 and 1R01AI104848 to B.D.B., and CA154947A, AI10008 and AI089987 to M.M.), the Beatriu de Pinós program (J.A.), and the Juvenile Diabetes Research Foundation (J.A.).

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Authors and Affiliations

  1. Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, New York, USA

    Judith Agudo, Albert Ruzo, Navpreet Tung, Alessia Baccarini & Brian D Brown

  2. Department of Oncological Sciences, Icahn School of Medicine at Mount Sinai, New York, New York, USA

    Hélène Salmon, Marylène Leboeuf, Daigo Hashimoto, Christian Becker & Miriam Merad

  3. Mount Sinai Immunology Institute, Icahn School of Medicine at Mount Sinai, New York, New York, USA

    Hélène Salmon, Marylène Leboeuf, Daigo Hashimoto, Christian Becker, Miriam Merad & Brian D Brown

  4. Department of Biochemistry, State University of New York at Buffalo, Buffalo, New York, USA

    Lee-Ann Garrett-Sinha

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Contributions

J.A. designed and did research and analyzed data; A.R., N.T., H.S., M.L., D.H., C.B., L.-A.G.-S. and A.B. did research; M.M. designed the project and analyzed data; and B.D.B. designed and coordinated the project and analyzed data.

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Correspondence to Brian D Brown.

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Integrated supplementary information

Supplementary Figure 1 Kinetics of the innate response to CpG DNA in WT and Mir126−/− mice.

(a) Measurement of serum IFN-a at the indicated time points after injection of CpG-A 2216 (4 μg) + DOTAP. Results are the mean±SD (n=3). (b) Measurement of serum IL-6 at the indicated time points after injection of CpG-A 2216 (4 μg) + DOTAP. Results are the mean±SD (n=3).

Supplementary Figure 2 Tlr7 and Tlr9 expression across the immune system and relative miRNA expression in different DC subsets.

(a) Tlr7 and Tlr9 expression pattern across the immune system as determined by the Immgen Consortium. Expression of each gene was measured by Affymetrix gene array on 234 different populations of immune cells that were double sorted to high purity. Shown is the mean±SD (n≥3). Note that the stroma includes both lymphatic and blood endothelial cells isolated from mesenteric lymph node. (b) Table showing miRNAs with the largest difference in expression between pDCs and the other DC subsets studied. Expression was determined by qPCR after sorting the indicated cells. The average fold difference (n=3-5) in expression of the indicated miRNA in pDCs compared to CD4+ DCs, CD8+ DCs, and CD103+ DCs was calculated by the ΔΔCT method.

Supplementary Figure 3 Loss of miR-126 impairs pDC homeostasis.

Eight week old Mir126-/- mice and WT littermates were analyzed. Representative flow cytometry plotsare shown. (a) CD4+ DC and CD8+ DC and B cells in the spleen. (b) CD4+ T cells and CD8+ T cells in the spleen. (c) (i) Eosinophils (Eos), (ii) neutrophils (Neut) and (iii) Gr1hi monocytes (mo) and (iv) Gr1low mo in the spleen.

Supplementary Figure 4 pDC proliferation is not affected by the loss of miR-126.

pDCs were differentiated in vitro from BM progenitors of Mir126-/- mice or WT littermate controls. pDC proliferation was measured by staining the cells with eFluor670 dye, and measuring dye dilution by flow cytometry at the indicated time points. (a) Shown are representative histograms gated on alive pDCs (DAPI- CD11c+ B220+ PDCA1+ cells) and all alive cells in the culture (DAPI-). (b) The graphs present the mean±SD (n=3-4).

Supplementary Figure 5 VEGFR2 is expressed in pDCs and can be downregulated in Kdrfl mice by Itgax-Cre expression.

(a) Protein expression of surface VEGFR2 in splenic pDCs was measured by flow cytometry. Note that VEGFR2 positive cells exclusively correspond to SiglecH positive cells (pDCs). Representative plot is shown. (b) Specific deletion of VEGFR2 was generated in vivo by crossing Kdr-floxed mice with Itgax-Cre mice. Deletion of VEGFR2 in pDCs was assessed by staining and flow cytometry analysis analysis. CD11cint B220+ SiglecH+ PDCA1+ cells are shown in representative histograms (n=3). (c) pDCs were sorted from KDRfl/fl–Itgax-Cre mice, and littermate controls for RNA extraction. VEGFR2 (Kdr) expression was measured by quantitative PCR using primers specific for exon 3. Graphs present the mean±SD (n=3). (d) Analysis of pDC frequency in the bone marrow of 2-month old KDRfl/fl-Itgax-Cre mice and WT littermates. pDCs were defined as CD11cint B220+ SiglecH+ PDCA1+ cells. Shown is a representative flow cytometry analysis plot and a graph of the mean±SD (n=5).

Supplementary Figure 6 miR-126–VEGFR2 control of pDC survival and function operates through regulation of the PI(3)K-Akt-mTOR pathway.

(a) As pDCs differentiate from progenitors they start to express high levels of miR-126, which suppresses the translation of TSC1. Reduced expression of TSC1 enhances mTOR activity, which improves cell survival and TLR signaling. This results in more pDCs and increased type I interferon production. It also results in increased expression of VEGFR2. Signaling through VEGFR2 further suppresses TSC1 activity, and, this in turn, enhances mTOR activity. (b) Knockout of miR-126 leads to increased TSC1 expression, which suppresses mTOR activity. This leads to decreased VEGFR2 expression, which further increases TSC1 activity and reduces mTOR activity. The outcome is increased pDC apoptosis and weaker TLR signaling, which results in reduced numbers of pDCs and diminished type I interferon production.

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Predicted targets of miR-126. (XLSX 53 kb)

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Agudo, J., Ruzo, A., Tung, N. et al. The miR-126–VEGFR2 axis controls the innate response to pathogen-associated nucleic acids. Nat Immunol 15, 54–62 (2014). https://doi.org/10.1038/ni.2767

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