Orphan nuclear receptor NR4A1 regulates transforming growth factor-β signaling and fibrosis


Mesenchymal responses are an essential aspect of tissue repair. Failure to terminate this repair process correctly, however, results in fibrosis and organ dysfunction. Therapies that block fibrosis and restore tissue homeostasis are not yet available for clinical use. Here we characterize the nuclear receptor NR4A1 as an endogenous inhibitor of transforming growth factor-β (TGF-β) signaling and as a potential target for anti-fibrotic therapies. NR4A1 recruits a repressor complex comprising SP1, SIN3A, CoREST, LSD1, and HDAC1 to TGF-β target genes, thereby limiting pro-fibrotic TGF-β effects. Even though temporary upregulation of TGF-β in physiologic wound healing induces NR4A1 expression and thereby creates a negative feedback loop, the persistent activation of TGF-β signaling in fibrotic diseases uses AKT- and HDAC-dependent mechanisms to inhibit NR4A1 expression and activation. Small-molecule NR4A1 agonists can overcome this lack of active NR4A1 and inhibit experimentally-induced skin, lung, liver, and kidney fibrosis in mice. Our data demonstrate a regulatory role of NR4A1 in TGF-β signaling and fibrosis, providing the first proof of concept for targeting NR4A1 in fibrotic diseases.

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Figure 1: TGF-β–dependent overexpression of pan-NR4A1.
Figure 2: Deficiency of NR4A1 exacerbates fibrosis.
Figure 3: NR4A1 inhibits TGF-β signaling.
Figure 4: NR4A1 inhibits collagen synthesis.
Figure 5: Inactivation of NR4A1 in fibrosis.
Figure 6: Csn-B for the treatment of fibrosis.


  1. 1

    Wynn, T.A. Cellular and molecular mechanisms of fibrosis. J. Pathol. 214, 199–210 (2008).

  2. 2

    Massagué, J. TGFβ signalling in context. Nat. Rev. Mol. Cell Biol. 13, 616–630 (2012).

  3. 3

    Gurtner, G.C., Werner, S., Barrandon, Y. & Longaker, M.T. Wound repair and regeneration. Nature 453, 314–321 (2008).

  4. 4

    Gabrielli, A., Avvedimento, E.V. & Krieg, T. Scleroderma. N. Engl. J. Med. 360, 1989–2003 (2009).

  5. 5

    Duffield, J.S., Lupher, M., Thannickal, V.J. & Wynn, T.A. Host responses in tissue repair and fibrosis. Annu. Rev. Pathol. 8, 241–276 (2013).

  6. 6

    Beyer, C. et al. Activation of pregnane X receptor inhibits experimental dermal fibrosis. Ann. Rheum. Dis. 72, 621–625 (2013).

  7. 7

    Zerr, P. et al. Vitamin D receptor regulates TGF-β signalling in systemic sclerosis. Ann. Rheum. Dis. doi:10.1136/annrheumdis-2013-204378 (21 January 2014).

  8. 8

    Wu, M. et al. Rosiglitazone abrogates bleomycin-induced scleroderma and blocks profibrotic responses through peroxisome proliferator-activated receptor-γ. Am. J. Pathol. 174, 519–533 (2009).

  9. 9

    Milbrandt, J. Nerve growth factor induces a gene homologous to the glucocorticoid receptor gene. Neuron 1, 183–188 (1988).

  10. 10

    Maxwell, M.A. et al. Nur77 regulates lipolysis in skeletal muscle cells. Evidence for cross-talk between the beta-adrenergic and an orphan nuclear hormone receptor pathway. J. Biol. Chem. 280, 12573–12584 (2005).

  11. 11

    Pei, L. et al. NR4A orphan nuclear receptors are transcriptional regulators of hepatic glucose metabolism. Nat. Med. 12, 1048–1055 (2006).

  12. 12

    Fassett, M.S., Jiang, W., D'Alise, A.M., Mathis, D. & Benoist, C. Nuclear receptor Nr4a1 modulates both regulatory T-cell (Treg) differentiation and clonal deletion. Proc. Natl. Acad. Sci. USA 109, 3891–3896 (2012).

  13. 13

    Hanna, R.N. et al. The transcription factor NR4A1 (Nur77) controls bone marrow differentiation and the survival of Ly6C monocytes. Nat. Immunol. 12, 778–785 (2011).

  14. 14

    Zeng, H. et al. Orphan nuclear receptor TR3/Nur77 regulates VEGF-A-induced angiogenesis through its transcriptional activity. J. Exp. Med. 203, 719–729 (2006).

  15. 15

    Arkenbout, E.K. et al. Protective function of transcription factor TR3 orphan receptor in atherogenesis: decreased lesion formation in carotid artery ligation model in TR3 transgenic mice. Circulation 106, 1530–1535 (2002).

  16. 16

    Han, Y.H. et al. Regulation of Nur77 nuclear export by c-Jun N-terminal kinase and Akt. Oncogene 25, 2974–2986 (2006).

  17. 17

    Chen, H.Z. et al. The orphan receptor TR3 suppresses intestinal tumorigenesis in mice by downregulating Wnt signalling. Gut 61, 714–724 (2012).

  18. 18

    Zhan, Y. et al. Cytosporone B is an agonist for nuclear orphan receptor Nur77. Nat. Chem. Biol. 4, 548–556 (2008).

  19. 19

    Uhl, M. et al. SD-208, a novel transforming growth factor beta receptor I kinase inhibitor, inhibits growth and invasiveness and enhances immunogenicity of murine and human glioma cells in vitro and in vivo. Cancer Res. 64, 7954–7961 (2004).

  20. 20

    Datta, P.K., Blake, M.C. & Moses, H.L. Regulation of plasminogen activator inhibitor-1 expression by transforming growth factor-beta -induced physical and functional interactions between smads and Sp1. J. Biol. Chem. 275, 40014–40019 (2000).

  21. 21

    Loeys, B.L. et al. Mutations in fibrillin-1 cause congenital scleroderma: stiff skin syndrome. Sci. Transl. Med. 2, 23ra20 (2010).

  22. 22

    Pascual, G. & Glass, C.K. Nuclear receptors versus inflammation: mechanisms of transrepression. Trends Endocrinol. Metab. 17, 321–327 (2006).

  23. 23

    Lee, S.O. et al. Inactivation of the orphan nuclear receptor TR3/Nur77 inhibits pancreatic cancer cell and tumor growth. Cancer Res. 70, 6824–6836 (2010).

  24. 24

    Poncelet, A.C. & Schnaper, H.W. Sp1 and Smad proteins cooperate to mediate transforming growth factor-beta 1-induced alpha 2(I) collagen expression in human glomerular mesangial cells. J. Biol. Chem. 276, 6983–6992 (2001).

  25. 25

    Zhou, F. et al. Nuclear receptor NR4A1 promotes breast cancer invasion and metastasis by activating TGF-β signalling. Nat. Commun. 5, 3388 (2014).

  26. 26

    Sonnylal, S. et al. Postnatal induction of transforming growth factor beta signaling in fibroblasts of mice recapitulates clinical, histologic, and biochemical features of scleroderma. Arthritis Rheum. 56, 334–344 (2007).

  27. 27

    Galiano, R.D., Michaels, J.t., Dobryansky, M., Levine, J.P. & Gurtner, G.C. Quantitative and reproducible murine model of excisional wound healing. Wound Repair Regen. 12, 485–492 (2004).

  28. 28

    Wong, V.W., Sorkin, M., Glotzbach, J.P., Longaker, M.T. & Gurtner, G.C. Surgical approaches to create murine models of human wound healing. J. Biomed. Biotechnol. 2011, 969618 (2011).

  29. 29

    Lin, B. et al. Conversion of Bcl-2 from protector to killer by interaction with nuclear orphan receptor Nur77/TR3. Cell 116, 527–540 (2004).

  30. 30

    Mullican, S.E. et al. Abrogation of nuclear receptors Nr4a3 and Nr4a1 leads to development of acute myeloid leukemia. Nat. Med. 13, 730–735 (2007).

  31. 31

    Beyer, C. & Distler, J.H. Tyrosine kinase signaling in fibrotic disorders: Translation of basic research to human disease. Biochim. Biophys. Acta 7, 897–904 (2013).

  32. 32

    Lee, S.O. et al. The nuclear receptor TR3 regulates mTORC1 signaling in lung cancer cells expressing wild-type p53. Oncogene 31, 3265–3276 (2012).

  33. 33

    Li, H. et al. Cytochrome c release and apoptosis induced by mitochondrial targeting of nuclear orphan receptor TR3. Science 289, 1159–1164 (2000).

  34. 34

    Thompson, J. & Winoto, A. During negative selection, Nur77 family proteins translocate to mitochondria where they associate with Bcl-2 and expose its proapoptotic BH3 domain. J. Exp. Med. 205, 1029–1036 (2008).

  35. 35

    Liu, J.J. et al. A unique pharmacophore for activation of the nuclear orphan receptor Nur77 in vivo and in vitro. Cancer Res. 70, 3628–3637 (2010).

  36. 36

    Lee, S.L. et al. Unimpaired thymic and peripheral T cell death in mice lacking the nuclear receptor NGFI-B (Nur77). Science 269, 532–535 (1995).

  37. 37

    Dees, C. et al. Platelet-derived serotonin links vascular disease and tissue fibrosis. J. Exp. Med. 208, 961–972 (2011).

  38. 38

    Wieser, R., Wrana, J.L. & Massague, J. GS domain mutations that constitutively activate T beta R-I, the downstream signaling component in the TGF-β receptor complex. EMBO J. 14, 2199–2208 (1995).

  39. 39

    Akhmetshina, A. et al. Activation of canonical Wnt signalling is required for TGF-β-mediated fibrosis. Nat. Commun. 3, 735 (2012).

  40. 40

    Hecker, L. et al. NADPH oxidase-4 mediates myofibroblast activation and fibrogenic responses to lung injury. Nat. Med. 15, 1077–1081 (2009).

  41. 41

    Hernández-Gea, V. et al. Autophagy releases lipid that promotes fibrogenesis by activated hepatic stellate cells in mice and in human tissues. Gastroenterology 142, 938–946 (2012).

  42. 42

    Moles, A. et al. A TLR2/S100A9/CXCL-2 signaling network is necessary for neutrophil recruitment in acute and chronic liver injury in the mouse. J. Hepatol. 60, 782–791 (2014).

  43. 43

    Bechtel, W. et al. Methylation determines fibroblast activation and fibrogenesis in the kidney. Nat. Med. 16, 544–550 (2010).

  44. 44

    Zeisberg, M. et al. BMP-7 counteracts TGF-β1-induced epithelial-to-mesenchymal transition and reverses chronic renal injury. Nat. Med. 9, 964–968 (2003).

  45. 45

    Wendling, O., Bornert, J.M., Chambon, P. & Metzger, D. Efficient temporally-controlled targeted mutagenesis in smooth muscle cells of the adult mouse. Genesis 47, 14–18 (2009).

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We thank D. Metzger and H. Ichinose for kindly providing us with Nr4a1fl/fl and αSma-Cre-ER mice. We thank M. Pascual Mate, R. Kleinlein, K. Dreißigacker and S. Fritz for excellent technical support. We also thank T. Kireva and E. Gianchecchi for their support in the study. We thank S. Wirtz and A. Weidemann for the expertise in experimental hepatic and kidney fibrosis. All figure drawings were adapted from Servier Medical Art. This study was supported by grants A40, A57, J20 and J29 of the Interdisciplinary Center for Clinical Research (IZKF) in Erlangen, individual grants from the German Research Foundation (Grants DI 1537/4-1, DI 1537/5-1, DI 1537/8-1, DI 1537/9-1 (Heisenberg Professorship), BE 5191/1-1, AK 144/1-1 and SCHE 1583/7-1), the SPP1468-IMMUNOBONE program of the German Research Foundation, the IMI-funded project BTCURE, the Metarthros project of the German Ministry of Education and Sciences and the project 00023728 from the Ministry of Health of the Czech Republic for conceptual development and SVV260031.

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K.P.-Z., A.D. and J.H.W.D. designed the research; K.P.-Z., P.Z., A.D., R.M., J.F., J.H., M.T., B.G.F., C.S., C.D., D. Metzger, and C.B. performed the research; K.P.-Z., P.Z., A.D., D. Mielenz, G.K., O.D., G.S. and J.H.W.D. analyzed the data; K.P.-Z., C.B., O.D., G.S. and J.H.W.D. wrote the manuscript.

Correspondence to Jörg H W Distler.

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Competing interests

O. Distler has consultancy relationships and/or has received research funding from Actelion, Pfizer, Ergonex, BMS, Sanofi-Aventis, United BioSource Corporation, Roche/Genentech, Medac, Biovitrium, Boehringer Ingelheim, Novartis, 4D Science, Active Biotec, Bayer, Sinoxa, Serodapharm, EpiPharm, GSK, Pharmacyclics and Biogen. J.H.W. Distler has consultancy relationships and/or has received research funding from Actelion, Active Biotech, Array Biopharma, Bayer Pharma, Boehringer Ingelheim, Celgene, GSK, JB Therapeutics, Novartis, Sanofi-Aventis, Sigma Tau and UCB in the area of potential treatments of SSc and is stock owner of 4D Science GmbH. 4D Science has a research cooperation with KaroBio on nuclear receptors as targets for anti-fibrotic therapies. All other authors declare no competing financial interests.

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Palumbo-Zerr, K., Zerr, P., Distler, A. et al. Orphan nuclear receptor NR4A1 regulates transforming growth factor-β signaling and fibrosis. Nat Med 21, 150–158 (2015). https://doi.org/10.1038/nm.3777

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