Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Colchicine acts selectively in the liver to induce hepatokines that inhibit myeloid cell activation

Nature Metabolism volume 3, pages 513–522 (2021)Cite this article

Subjects

A Publisher Correction to this article was published on 05 May 2021

This article has been updated

Abstract

Colchicine has served as a traditional medicine for millennia and remains widely used to treat inflammatory and other disorders. Colchicine binds tubulin and depolymerizes microtubules, but it remains unclear how this mechanism blocks myeloid cell recruitment to inflamed tissues. Here we show that colchicine inhibits myeloid cell activation via an indirect mechanism involving the release of hepatokines. We find that a safe dose of colchicine depolymerizes microtubules selectively in hepatocytes but not in circulating myeloid cells. Mechanistically, colchicine triggers Nrf2 activation in hepatocytes, leading to secretion of anti-inflammatory hepatokines, including growth differentiation factor 15 (GDF15). Nrf2 and GDF15 are required for the anti-inflammatory action of colchicine in vivo. Plasma from colchicine-treated mice inhibits inflammatory signalling in myeloid cells in a GDF15-dependent manner, by positive regulation of SHP-1 (PTPN6) phosphatase, although the precise molecular identities of colchicine-induced GDF15 and its receptor require further characterization. Our work shows that the efficacy and safety of colchicine depend on its selective action on hepatocytes, and reveals a new axis of liver–myeloid cell communication. Plasma GDF15 levels and myeloid cell SHP-1 activity may be useful pharmacodynamic biomarkers of colchicine action.

Access through your institution
Buy or subscribe

This is a preview of subscription content, access via your institution

Access options

Access through your institution

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Fig. 1: Colchicine induces anti-inflammatory mediators and selectively targets hepatocytes.
Fig. 2: Colchicine activates Nrf2–Keap1 signalling.
Fig. 3: Nrf2 protects the liver against colchicine.
Fig. 4: Colchicine-induced GDF15 is an anti-inflammatory hepatokine.
Fig. 5: SHP-1 phosphatase mediates the anti-inflammatory effects of colchicine-induced hepatokines on myeloid cells.

Similar content being viewed by others

Data availability

The data that support the findings of this study are available from the corresponding authors on reasonable request. Source data are provided with this paper.

Change history

References

  1. Medzhitov, R. Origin and physiological roles of inflammation. Nature 454, 428–435 (2008).

    Article  CAS  PubMed  Google Scholar 

  2. Dinarello, C. A. Anti-inflammatory agents: present and future. Cell 140, 935–950 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Dasgeb, B. et al. Colchicine: an ancient drug with novel applications. Br. J. Dermatol. 178, 350–356 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Nidorf, S. M., Eikelboom, J. W., Budgeon, C. A. & Thompson, P. L. Low-dose colchicine for secondary prevention of cardiovascular disease. J. Am. Coll. Cardiol. 61, 404–410 (2013).

    Article  CAS  PubMed  Google Scholar 

  5. Tardif, J. C. et al. Efficacy and safety of low-dose colchicine after myocardial infarction. N. Engl. J. Med. 381, 2497–2505 (2019).

  6. Deftereos, S. G. et al. Effect of colchicine vs standard care on cardiac and inflammatory biomarkers and clinical outcomes in patients hospitalized with coronavirus disease 2019: the GRECCO-19 randomized clinical trial. JAMA Netw. Open 3, e2013136 (2020).

    Article  PubMed  PubMed Central  Google Scholar 

  7. Finkelstein, Y. et al. Colchicine poisoning: the dark side of an ancient drug. Clin. Toxicol. (Phila.) 48, 407–414 (2010).

    Article  CAS  Google Scholar 

  8. Weisenberg, R. C., Borisy, G. G. & Taylor, E. W. The colchicine-binding protein of mammalian brain and its relation to microtubules. Biochemistry 7, 4466–4479 (1968).

    Article  CAS  PubMed  Google Scholar 

  9. Dinarello, C. A. et al. Effect of prophylactic colchicine therapy on leukocyte function in patients with familial Mediterranean fever. Arthritis Rheumatol. 19, 618–622 (1976).

    Article  CAS  Google Scholar 

  10. Caner, J. E. Colchicine inhibition of chemotaxis. Arthritis Rheumatol. 8, 757–764 (1965).

    Article  CAS  Google Scholar 

  11. Martinon, F., Petrilli, V., Mayor, A., Tardivel, A. & Tschopp, J. Gout-associated uric acid crystals activate the NALP3 inflammasome. Nature 440, 237–241 (2006).

    Article  CAS  PubMed  Google Scholar 

  12. Thomas, G., Girre, C., Scherrmann, J. M., Francheteau, P. & Steimer, J. L. Zero-order absorption and linear disposition of oral colchicine in healthy volunteers. Eur. J. Clin. Pharm. 37, 79–84 (1989).

    Article  CAS  Google Scholar 

  13. Florian, S. & Mitchison, T. J. Anti-microtubule drugs. Methods Mol. Biol. 1413, 403–421 (2016).

    Article  CAS  PubMed  Google Scholar 

  14. Hunter, A. L. & Klaassen, C. D. Biliary excretion of colchicine. J. Pharmacol. Exp. Ther. 192, 605–617 (1975).

    CAS  PubMed  Google Scholar 

  15. Ghorpade, D. S. et al. Hepatocyte-secreted DPP4 in obesity promotes adipose inflammation and insulin resistance. Nature 555, 673–677 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Ferguson, F. C. Jr. Colchicine. I. General pharmacology. J. Pharmacol. Exp. Ther. 106, 261–270 (1952).

    CAS  PubMed  Google Scholar 

  17. Terkeltaub, R. A. et al. High versus low dosing of oral colchicine for early acute gout flare: twenty-four-hour outcome of the first multicenter, randomized, double-blind, placebo-controlled, parallel-group, dose-comparison colchicine study. Arthritis Rheumatol. 62, 1060–1068 (2010).

    Article  CAS  Google Scholar 

  18. Wang, T. H. et al. Microtubule-interfering agents activate c-Jun N-terminal kinase/stress-activated protein kinase through both Ras and apoptosis signal-regulating kinase pathways. J. Biol. Chem. 273, 4928–4936 (1998).

    Article  CAS  PubMed  Google Scholar 

  19. Cereghini, S. Liver-enriched transcription factors and hepatocyte differentiation. FASEB J. 10, 267–282 (1996).

    Article  CAS  PubMed  Google Scholar 

  20. Faber, K. N., Muller, M. & Jansen, P. L. Drug transport proteins in the liver. Adv. Drug Deliv. Rev. 55, 107–124 (2003).

    Article  CAS  PubMed  Google Scholar 

  21. Bergen, L. G. & Borisy, G. G. Tubulin-colchicine complex inhibits microtubule elongation at both plus and minus ends. J. Biol. Chem. 258, 4190–4194 (1983).

    Article  CAS  PubMed  Google Scholar 

  22. Magupalli, V. G. et al. HDAC6 mediates an aggresome-like mechanism for NLRP3 and pyrin inflammasome activation. Science 369, eaas8995 (2020).

  23. Ozen, S. et al. EULAR recommendations for the management of familial Mediterranean fever. Ann. Rheum. Dis. 75, 644–651 (2016).

    Article  CAS  PubMed  Google Scholar 

  24. Cuadrado, A. et al. Therapeutic targeting of the NRF2 and KEAP1 partnership in chronic diseases. Nat. Rev. Drug Discov. 18, 295–317 (2019).

    Article  CAS  PubMed  Google Scholar 

  25. Komatsu, M. et al. The selective autophagy substrate p62 activates the stress responsive transcription factor Nrf2 through inactivation of Keap1. Nat. Cell Biol. 12, 213–223 (2010).

    Article  CAS  PubMed  Google Scholar 

  26. Bae, S. H. et al. Sestrins activate Nrf2 by promoting p62-dependent autophagic degradation of Keap1 and prevent oxidative liver damage. Cell Metab. 17, 73–84 (2013).

    Article  CAS  PubMed  Google Scholar 

  27. Ichimura, Y. et al. Phosphorylation of p62 activates the Keap1-Nrf2 pathway during selective autophagy. Mol. Cell 51, 618–631 (2013).

    Article  CAS  PubMed  Google Scholar 

  28. Johansen, T. & Lamark, T. Selective autophagy mediated by autophagic adapter proteins. Autophagy 7, 279–296 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Igarashi, Y. et al. Open TG-GATEs: a large-scale toxicogenomics database. Nucleic Acids Res. 43, D921–D927 (2015).

    Article  CAS  PubMed  Google Scholar 

  30. Hsu, J. Y. et al. Non-homeostatic body weight regulation through a brainstem-restricted receptor for GDF15. Nature 550, 255–259 (2017).

    Article  PubMed  Google Scholar 

  31. Kim, K. H. et al. Growth differentiation factor 15 ameliorates nonalcoholic steatohepatitis and related metabolic disorders in mice. Sci. Rep. 8, 6789 (2018).

    Article  PubMed  PubMed Central  Google Scholar 

  32. de Jager, S. C. et al. Growth differentiation factor 15 deficiency protects against atherosclerosis by attenuating CCR2-mediated macrophage chemotaxis. J. Exp. Med. 208, 217–225 (2011).

    Article  PubMed  PubMed Central  Google Scholar 

  33. Kempf, T. et al. The transforming growth factor-beta superfamily member growth-differentiation factor-15 protects the heart from ischemia/reperfusion injury. Circ. Res. 98, 351–360 (2006).

    Article  CAS  PubMed  Google Scholar 

  34. Luan, H. H. et al. GDF15 Is an Inflammation-Induced central mediator of tissue tolerance. Cell 178, 1231–1244 e1211 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Yatsuga, S. et al. Growth differentiation factor 15 as a useful biomarker for mitochondrial disorders. Ann. Neurol. 78, 814–823 (2015).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Wollert, K. C., Kempf, T. & Wallentin, L. Growth differentiation factor 15 as a biomarker in cardiovascular disease. Clin. Chem. 63, 140–151 (2017).

    Article  CAS  PubMed  Google Scholar 

  37. Johnen, H. et al. Tumor-induced anorexia and weight loss are mediated by the TGF-beta superfamily cytokine MIC-1. Nat. Med. 13, 1333–1340 (2007).

    Article  CAS  PubMed  Google Scholar 

  38. Tanaka, T. et al. Plasma proteomic signature of age in healthy humans. Aging Cell 17, e12799 (2018).

    Article  PubMed  PubMed Central  Google Scholar 

  39. Fejzo, M. S. et al. Placenta and appetite genes GDF15 and IGFBP7 are associated with hyperemesis gravidarum. Nat. Commun. 9, 1178 (2018).

    Article  PubMed  PubMed Central  Google Scholar 

  40. Demidowich, A. P. et al. Colchicine’s effects on metabolic and inflammatory molecules in adults with obesity and metabolic syndrome: results from a pilot randomized controlled trial. Int. J. Obes. 44, 1793–1799 (2020).

    Article  CAS  Google Scholar 

  41. Nair, J. K. et al. Multivalent N-acetylgalactosamine-conjugated siRNA localizes in hepatocytes and elicits robust RNAi-mediated gene silencing. J. Am. Chem. Soc. 136, 16958–16961 (2014).

    Article  CAS  PubMed  Google Scholar 

  42. Coll, A. P. et al. GDF15 mediates the effects of metformin on body weight and energy balance. Nature 578, 444–448 (2020).

    Article  CAS  PubMed  Google Scholar 

  43. Mocsai, A., Ruland, J. & Tybulewicz, V. L. The SYK tyrosine kinase: a crucial player in diverse biological functions. Nat. Rev. Immunol. 10, 387–402 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Veillette, A., Latour, S. & Davidson, D. Negative regulation of immunoreceptor signaling. Annu. Rev. Immunol. 20, 669–707 (2002).

    Article  CAS  PubMed  Google Scholar 

  45. Abram, C. L. & Lowell, C. A. Shp1 function in myeloid cells. J. Leukoc. Biol. 102, 657–675 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Kundu, S. et al. Novel SHP-1 inhibitors tyrosine phosphatase inhibitor-1 and analogs with preclinical anti-tumor activities as tolerated oral agents. J. Immunol. 184, 6529–6536 (2010).

    Article  CAS  PubMed  Google Scholar 

  47. Hsiao, E. C. et al. Characterization of growth-differentiation factor 15, a transforming growth factor beta superfamily member induced following liver injury. Mol. Cell. Biol. 20, 3742–3751 (2000).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Yang, L. et al. GFRAL is the receptor for GDF15 and is required for the anti-obesity effects of the ligand. Nat. Med. 23, 1158–1166 (2017).

    Article  CAS  PubMed  Google Scholar 

  49. Constam, D. B. Regulation of TGFbeta and related signals by precursor processing. Semin. Cell Dev. Biol. 32, 85–97 (2014).

    Article  CAS  PubMed  Google Scholar 

  50. Derynck, R. & Budi, E. H. Specificity, versatility, and control of TGF-beta family signaling. Sci. Signal. 12, eaav5183 (2019).

  51. Hinck, A. P., Mueller, T. D. & Springer, T. A. Structural biology and evolution of the TGF-beta family. Cold Spring Harb. Perspect. Biol. https://doi.org/10.1101/cshperspect.a022103 (2016).

  52. Fordham, J. N., Kirwan, J., Cason, J. & Currey, H. L. Prolonged reduction in polymorphonuclear adhesion following oral colchicine. Ann. Rheum. Dis. 40, 605–608 (1981).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  53. Martinez, G. J. et al. Colchicine acutely suppresses local cardiac production of inflammatory cytokines in patients with an acute coronary syndrome. J. Am. Heart Assoc. 4, e002128 (2015).

    Article  PubMed  PubMed Central  Google Scholar 

  54. Robinson, K. P. & Chan, J. J. Colchicine in dermatology: a review. Australas. J. Dermatol. 59, 278–285 (2018).

    Article  PubMed  Google Scholar 

  55. Weng, J. H. et al. Pregnenolone activates CLIP-170 to promote microtubule growth and cell migration. Nat. Chem. Biol. 9, 636–642 (2013).

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank S.-J. Lee for the Gdf15−/− mice and J. Kagan for immortalized BMDM cells. The GDF15-blocking antibody was a kind gift from H. Tian of NGM Biopharmaceuticals. We thank W.-T. Kuo of Harvard Medical School for assistance with tissue immunostaining. We thank M. Zhou of NGM Biopharmaceuticals for assistance with animal experiments. This work was supported by grants from the National Institutes of Health (no. GM131753 to T.J.M) and the American Heart Association (no. 8POST34080251 to J.-H.W.).

Author information

Authors and Affiliations

  1. Department of Systems Biology, Blavatnik Institute, Harvard Medical School, Boston, MA, USA

    Jui-Hsia Weng, Peter David Koch, Kenichi Shimada, Ruomu Jiang & Timothy J. Mitchison

  2. Center for Systems Biology, Massachusetts General Hospital, Boston, MA, USA

    Peter David Koch

  3. NGM Biopharmaceuticals, South San Francisco, CA, USA

    Harding H. Luan, Iris Ngan & Richard Ventura

  4. Alnylam Pharmaceuticals, Inc., Cambridge, MA, USA

    Ho-Chou Tu

Authors
  1. Jui-Hsia Weng
    View author publications

    Search author on:PubMed Google Scholar

  2. Peter David Koch
    View author publications

    Search author on:PubMed Google Scholar

  3. Harding H. Luan
    View author publications

    Search author on:PubMed Google Scholar

  4. Ho-Chou Tu
    View author publications

    Search author on:PubMed Google Scholar

  5. Kenichi Shimada
    View author publications

    Search author on:PubMed Google Scholar

  6. Iris Ngan
    View author publications

    Search author on:PubMed Google Scholar

  7. Richard Ventura
    View author publications

    Search author on:PubMed Google Scholar

  8. Ruomu Jiang
    View author publications

    Search author on:PubMed Google Scholar

  9. Timothy J. Mitchison
    View author publications

    Search author on:PubMed Google Scholar

Contributions

J.-H.W. designed and conducted the experiments and wrote the manuscript. P.D.K. performed PK-PD modelling and image quantification. H.H.L. and I.N. examined GFRAL neutralization with the peritonitis assay. H.-C.T. generated hepatocyte-specific siRNAs. K.S. and R.J. examined the Open TG-GATEs database. R.V. tested the activity of recombinant GDF15. T.J.M. oversaw the execution of the project and wrote the manuscript.

Corresponding authors

Correspondence to Jui-Hsia Weng or Timothy J. Mitchison.

Ethics declarations

Competing interests

H.-C.T. was an employee of Alnylam Pharmaceuticals when the work was done. H.H.L., I.N. and R.V. were employees of NGM Biopharmaceuticals when the work was done. The other authors declare no competing interests.

Additional information

Peer review information Primary Handling Editor: Christoph Schmitt.

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Extended data

Extended Data Fig. 1 Selection of colchicine doses for mouse treatment.

a, Dose translation from humans to animal studies based on clinical guidelines. The colchicine doses for mouse via the oral or i.p. routes were calculated according to the guidelines from the Food and Drug Administration and the European League Against Rheumatism, and were listed in the table. i.p., intraperitoneal. i.v., intravascular. b-f, Safety analysis of colchicine based on measurement of gut toxicity. Diarrhea is the dose-limiting toxicity in man. Mice received either vehicle, colchicine at 0.4 mg/kg, or 2.4 mg/kg. b, Intestines were harvested 6 hours after vehicle or colchicine treatment. Scale bars, 1 cm. c, The physiological severity score for diarrhea. d, Colchicine at 0.4 mg/kg did not affect the region of intestine with feces, but 2.4 mg/kg reduced it. Each dot represents one mouse. e, Detection of fecal water content by measuring weight of feces before and after drying. f, Colchicine at 0.4 mg/kg did not affect the fecal water content. Data are represented as mean ± s.d. Two-sided t-tests were used for statistical analysis. A single colchicine dose of 0.4mg/mg i.p. was used in all subsequent experiments unless otherwise indicated.

Extended Data Fig. 2 Colchicine at a safe dose selectively targets hepatocytes.

Additional images related to Fig. 1g. Microtubules in the livers of mice treated with a, vehicle or b, 0.4 mg/kg colchicine. Livers were collected 6 hours after treatment. Colchicine selectively depolymerized microtubules in hepatocytes, identified by HNF4+. The boxed areas are magnified and shown at the bottom. Liver cells from 4 livers with vehicle or colchicine treatment respectively. Scale bars, 10 μm.

Extended Data Fig. 3 Colchicine at a safe dose does not damage microtubules in circulating blood cells.

Six hours after treatment of vehicle or colchicine (0.4 mg/kg or 2.4 mg/kg), the whole blood samples were collected and fixed directly. Microtubules were visualized by immunofluorescence and imaged by confocal microscopy. a, Representative images of microtubules in circulating blood cells. b, Quantification of microtubule staining intensity. Individual cells exhibited a wide range of microtubule intensities that were similar following vehicle or 0.4 mg/kg colchicine (the safe dose). Microtubules were depolymerized in cells from mice receiving 2.4 mg/kg (the toxic dose). AU, arbitrary unit. Scale bar, 5 μm.

Extended Data Fig. 4 Colchicine at a toxic dose targets all cell types in the liver.

Microtubules in a, vehicle- or b, 2.4 mg/kg colchicine-treated livers were visualized, and c, intensities of microtubules were quantified. Microtubules were depolymerized in all cell types after treatment of colchicine at 2.4 mg/kg. The boxed areas are magnified and shown at the bottom. AU, arbitrary unit. Scale bars, 10 μm.

Extended Data Fig. 5 Modeling of human pharmacokinetics-pharmacodynamics (PK-PD) of colchicine.

a, Simulation diagram of two-compartment PK-PD model, based on human pharmacology data from Thomas, G. et al. Two blue boxes represent the central and peripheral compartments, which correspond to plasma and myeloid cell respectively. Light green boxes correspond to reaction species (colchicine concentration in plasma or myeloid cell), and yellow circles correspond to reactions (absorption, urinary elimination, central elimination, and transfer between plasma and myeloid cell). Colchicine absorption takes place from t = 0.22 to t = 1 hour. Tubulin-colchicine binding biochemistry was ignored and all colchicine in the myeloid cells was assumed to bind to tubulin. Fraction bound was calculated based on equations listed in Methods. b, Constants for modeling. c, Human PK-PD modeling of colchicine supported the indirect action of colchicine. The blue line represents colchicine concentrations in plasma over time after colchicine dosing. Magenta lines show the fraction of colchicine-bound tubulin in myeloid cells. Dashed and dotted magenta lines show unbound colchicine concentrated across the cell membrane by a factor of 10 and 100 respectively. The green line marks the 2% threshold of site occupancy for colchicine action.

Extended Data Fig. 6 GDF15 is a novel hepatokine induced by colchicine.

a, Volcano plot for the gene expression changes of rat livers comparing 3 hours after vehicle to colchicine treatment. Biological triplicate data are from the Open TG-GATEs database. Red and gray dots represent genes encoding secreted and non-secreted proteins respectively. GDF15 is highlighted in blue. P, probability value. b, Time course of GDF15 mRNA expression in the rat liver. Data are from Open TG-GATEs. 3 Mice per condition. c, GDF15 was induced in the liver but not kidney collected from colchicine-treated mice. d, ELISA analysis for measurement of plasma GDF15 in both mouse sexes. Induction was reproducibly stronger in males in the C57BL/6J mouse strain. Each dot represents one mouse. 5 Mice per condition. 3 Biological replicates. Data are represented as mean ± s.d. Two-sided t-tests were used for statistical analysis.

Source data

Extended Data Fig. 7 GDF15 is a novel anti-inflammatory hepatokine induced by Nrf2 signaling triggered by colchicine.

a, ELISA analysis for measurement of plasma GDF15 in wild-type and Nrf2 germline knockout (Nrf2−/−) mice. Each dot represents one mouse. b, Immunoblotting analysis revealed that induction of GDF15 in livers was blocked by treatment of hepatocyte-specific siRNA (GalNAc-siRNA) against Nrf2. Luc, luciferase. 3 Biological replicates. c, Activation of GDF15 promoter activity by Nrf2. HEK 293T cells were co-transfected with the luciferase reporter driven by the GDF15 promoter and the Nrf2 plasmid. Luciferase activities were determined from three samples. Data from three independent experiments. d, Immunoblotting analysis for detection of CHOP expression in livers collected at the indicated times after colchicine or Tunicamycin treatment. e, The temporal mRNA expression pattern of the integrated stress response regulators in the rat liver. Data are from Open TG-GATEs. 3 Mice per condition. f, The anti-inflammatory effect of colchicine in the MSU (monosodium urate crystals) peritonitis model depended on GDF15 induction in hepatocytes. Mice were challenged with 1 mg MSU. GalNAc-siRNA, hepatocyte-specific siRNA. Luc, luciferase. Each dot represents one mouse. Data are represented as mean ± s.d. Two-sided t-tests were used for statistical analysis.

Source data

Extended Data Fig. 8 Colchicine-induced GDF15 inhibits pro-IL-1β expression by activating the immuno-inhibitory signal of SHP-1 in primary neutrophils.

Ex vivo activation of neutrophils evaluated by the levels of (a-b) pro-IL-1β induction or (c-d) phosphorylated SHP-1. Cells were pre-incubated with plasma from the indicated mice and then activated with PMA. a, Induction of GDF15 specifically in hepatocytes was required for the anti-inflammatory activity of post-colchicine plasma, as assayed by pro-IL-1β induction. GalNAc-siLuc, hepatocyte-specific siRNA against luciferase. GalNAc-siGDF15, hepatocyte-specific siRNA against GDF15. b, The anti-inflammatory activity of post-colchicine plasma was blocked by two different clones of antibodies (#2 and #3) against GDF15. ctr, control antibody. veh plasma, plasma from vehicle-treated mice. Colc plasma, plasma from colchicine-treated mice. c, GDF15 neutralization by two clones of anti-GDF15 antibodies (#2 and #3) blocked plasma activity on stabilizing SHP-1 activation. d, Expression of colchicine-induced GDF15 specifically in hepatocytes was required for activating SHP-1. 3 Biological replicates.

Source data

Extended Data Fig. 9 Colchicine-induced GDF15 inhibits pro-IL-1β expression and mature IL-1β secretion from macrophages.

Ex vivo activation of immortalized BMDMs evaluated by the levels of (a-b) pro-IL-1β expression and (d-e) IL-1β secretion. a-b, Cells were pre-incubated with plasma from the indicated mice and then activated with Pam3CSK4. Expression of colchicine-induced GDF15 in hepatocytes was required for plasma activity on blocking pro-IL-1β expression. GalNAc-siLuc, hepatocyte-specific siRNA against luciferase. GalNAc-siGDF15, hepatocyte-specific siRNA against GDF15. c, Activation methods for measuring IL-1β production. d, Colchicine did not alter IL-1β production assayed with two activation methods. 5 Samples per condition over 3 biological replicates e, Colchicine-induced GDF15 was required for plasma activity on reducing IL-1β production. 4 Samples per condition over 3 biological replicates Data are represented as mean ± s.d. Two-sided t-tests were used for statistical analysis.

Source data

Extended Data Fig. 10 Canonical mature GDF15-GFRAL-RET signaling is not involved in the anti-inflammatory actions of colchicine.

a-c, and f, Ex vivo activation of neutrophils evaluated by the levels of pro-IL-1β expression. Inhibition of the GFRAL-RET signaling by a, RET inhibitor, SPP86 (SPP) and BBT594 (BTT), or b, a GFRAL neutralizing antibody (α-GFRAL) did not alter post-colchicine plasma activity on blocking pro-IL-1β expression. c, Post-colchicine plasma from wild-type and GFRAL germline knockout mice blocked pro-IL-1β expression. d, GFRAL inhibition by a GFRAL neutralizing antibody did not affect the in vivo anti-inflammatory effect of colchicine in the zymosan peritonitis assay. e, Efficacy of recombinant mature GDF15 (rGDF15) measured by the levels of phosphorylated ERK in a GFRAL-RET reporter cell model. 3 Samples per condition. Recombinant GDF15 was a disulfide-linked mature homodimer. Recombinant mature GDF15 did not affect f, ex vivo pro-IL-1β expression, and did not inhibit g, in vivo zymosan-induced neutrophil recruitment as colchicine. Each dot represents one mouse. Data are represented as mean ± s.d. Two-sided t-tests were used for statistical analysis.

Source data

Supplementary information

Source data

Source Data Fig. 1

Unprocessed immunoblots.

Source Data Fig. 1

Statistical source data.

Source Data Fig. 2

Unprocessed immunoblots.

Source Data Fig. 2

Statistical source data.

Source Data Fig. 3

Source data.

Source Data Fig. 4

Unprocessed immunoblots.

Source Data Fig. 4

Statistical source data.

Source Data Fig. 5

Unprocessed immunoblots.

Source Data Fig. 5

Statistical source data.

Source Data Extended Data Fig. 6

Unprocessed immunoblots.

Source Data Extended Data Fig. 7

Unprocessed immunoblots.

Source Data Extended Data Fig. 8

Unprocessed immunoblots.

Source Data Extended Data Fig. 9

Unprocessed immunoblots.

Source Data Extended Data Fig. 10

Unprocessed western blots.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Weng, JH., Koch, P.D., Luan, H.H. et al. Colchicine acts selectively in the liver to induce hepatokines that inhibit myeloid cell activation. Nat Metab 3, 513–522 (2021). https://doi.org/10.1038/s42255-021-00366-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue date:

  • DOI: https://doi.org/10.1038/s42255-021-00366-y

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing