Subjects

Abstract

Fructose is a major component of dietary sugar and its overconsumption exacerbates key pathological features of metabolic syndrome. The central fructose-metabolising enzyme is ketohexokinase (KHK), which exists in two isoforms: KHK-A and KHK-C, generated through mutually exclusive alternative splicing of KHK pre-mRNAs. KHK-C displays superior affinity for fructose compared with KHK-A and is produced primarily in the liver, thus restricting fructose metabolism almost exclusively to this organ. Here we show that myocardial hypoxia actuates fructose metabolism in human and mouse models of pathological cardiac hypertrophy through hypoxia-inducible factor 1α (HIF1α) activation of SF3B1 and SF3B1-mediated splice switching of KHK-A to KHK-C. Heart-specific depletion of SF3B1 or genetic ablation of Khk, but not Khk-A alone, in mice, suppresses pathological stress-induced fructose metabolism, growth and contractile dysfunction, thus defining signalling components and molecular underpinnings of a fructose metabolism regulatory system crucial for pathological growth.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    , , & Hypertension and heart failure: a dysfunction of systole, diastole or both? J. Hum. Hypertens. 23, 295–306 (2009)

  2. 2.

    et al. Disruption of coordinated cardiac hypertrophy and angiogenesis contributes to the transition to heart failure. J. Clin. Invest. 115, 2108–2118 (2005)

  3. 3.

    & Oxygen sensing by metazoans: the central role of the HIF hydroxylase pathway. Mol. Cell 30, 393–402 (2008)

  4. 4.

    Oxygen sensing, hypoxia-inducible factors, and disease pathophysiology. Annu. Rev. Pathol. 9, 47–71 (2014)

  5. 5.

    & Cardiac plasticity. N. Engl. J. Med. 358, 1370–1380 (2008)

  6. 6.

    et al. Tissue-specific splicing factor gene expression signatures. Nucleic Acids Res. 36, 4823–4832 (2008)

  7. 7.

    , & Systematic genome-wide annotation of spliceosomal proteins reveals differential gene family expansion. Genome Res. 16, 66–77 (2006)

  8. 8.

    , , & Large-scale proteomic analysis of the human spliceosome. Genome Res. 12, 1231–1245 (2002)

  9. 9.

    et al. Ketohexokinase: expression and localization of the principal fructose-metabolizing enzyme. J. Histochem. Cytochem. 57, 763–774 (2009)

  10. 10.

    et al. Activation of a HIF1α-PPARγ axis underlies the integration of glycolytic and lipid anabolic pathways in pathologic cardiac hypertrophy. Cell Metab. 9, 512–524 (2009)

  11. 11.

    et al. Phosphorylation of spliceosomal protein SAP 155 coupled with splicing catalysis. Genes Dev. 12, 1409–1414 (1998)

  12. 12.

    , & The spliceosome as a target of novel antitumour drugs. Nature Rev. Drug Discov. 11, 847–859 (2012)

  13. 13.

    , & Reduced fidelity of branch point recognition and alternative splicing induced by the anti-tumor drug spliceostatin A. Genes Dev. 25, 445–459 (2011)

  14. 14.

    et al. Disruption of SF3B1 results in deregulated expression and splicing of key genes and pathways in myelodysplastic syndrome hematopoietic stem and progenitor cells. Leukemia 29, 1092–1103 (2015)

  15. 15.

    & Aldose reductase and cardiovascular diseases, creating human-like diabetic complications in an experimental model. Circ. Res. 106, 1449–1458 (2010)

  16. 16.

    et al. Tracing compartmentalized NADPH metabolism in the cytosol and mitochondria of mammalian cells. Mol. Cell 55, 253–263 (2014)

  17. 17.

    et al. Ketohexokinase-dependent metabolism of fructose induces proinflammatory mediators in proximal tubular cells. J. Am. Soc. Nephrol. 20, 545–553 (2009)

  18. 18.

    et al. The ER UDPase ENTPD5 promotes protein N-glycosylation, the Warburg effect, and proliferation in the PTEN pathway. Cell 143, 711–724 (2010)

  19. 19.

    et al. Evidence for an alternative glycolytic pathway in rapidly proliferating cells. Science 329, 1492–1499 (2010)

  20. 20.

    et al. Intramyocardial lipid accumulation in the failing human heart resembles the lipotoxic rat heart. FASEB J. 18, 1692–1700 (2004)

  21. 21.

    et al. Both isoforms of ketohexokinase are dispensable for normal growth and development. Physiol. Genomics 42A, 235–243 (2010)

  22. 22.

    , , & Regulation of HPV16 E6 and MCL1 by SF3B1 inhibitor in head and neck cancer cells. Sci. Rep. 4, 6098 (2014)

  23. 23.

    , , , & HnRNP proteins controlled by c-Myc deregulate pyruvate kinase mRNA splicing in cancer. Nature 463, 364–368 (2010)

  24. 24.

    , & Understanding the Warburg effect: the metabolic requirements of cell proliferation. Science 324, 1029–1033 (2009)

  25. 25.

    , , , & Small amounts of fructose markedly augment net hepatic glucose uptake in the conscious dog. Diabetes 47, 867–873 (1998)

  26. 26.

    et al. Pyruvate Kinase M2 Is a PHD3-Stimulated Coactivator for Hypoxia-Inducible Factor 1. Cell 145, 732–744 (2011)

  27. 27.

    et al. Diabetes mellitus worsens diastolic left ventricular dysfunction in aortic stenosis through altered myocardial structure and cardiomyocyte stiffness. Circulation 124, 1151–1159 (2011)

  28. 28.

    , & Increased fructose concentrations in blood and urine in patients with diabetes. Diabetes Care 25, 353–357 (2002)

  29. 29.

    et al. Myocardial structure and function differ in systolic and diastolic heart failure. Circulation 113, 1966–1973 (2006)

  30. 30.

    et al. Combination of tumor necrosis factor-α ablation and matrix metalloproteinase inhibition prevents heart failure after pressure overload in tissue inhibitor of metalloproteinase-3 knock-out mice. Circ. Res. 97, 380–390 (2005)

  31. 31.

    et al. The Notch pathway controls fibrotic and regenerative repair in the adult heart. Eur. Heart J. 21, 2174–2185 (2014)

  32. 32.

    et al. Connexin43-dependent mechanism modulates renin secretion and hypertension. J. Clin. Invest. 116, 405–413 (2006)

  33. 33.

    , , & Simplified lentivirus vector production in protein-free media using polyethylenimine-mediated transfection. J. Virol. Methods 157, 113–121 (2009)

  34. 34.

    et al. Cre-lox-regulated conditional RNA interference from transgenes. Proc. Natl Acad. Sci. USA 101, 10380–10385 (2004)

  35. 35.

    , , & Regulation of hypoxia-inducible factor 1α is mediated by an O2-dependent degradation domain via the ubiquitin-proteasome pathway. Proc. Natl Acad. Sci. USA 95, 7987–7992 (1998)

  36. 36.

    et al. HIF1α deubiquitination by USP8 is essential for ciliogenesis in normoxia. EMBO Rep. 15, 77–85 (2014)

  37. 37.

    et al. A new L1446P mutation is responsible for impaired von Willebrand factor synthesis, structure, and function. J. Lab. Clin. Med. 144, 254–259 (2004)

  38. 38.

    et al. Dietary obesity-associated Hif1 activation in adipocytes restricts fatty acid oxidation and energy expenditure via suppression of the Sirt2-NAD+ system. Genes Dev. 26, 259–270 (2012)

  39. 39.

    et al. iCLIP reveals the function of hnRNP particles in splicing at individual nucleotide resolution. Nature Struct. Mol. Biol. 17, 909–915 (2010)

  40. 40.

    , , & Establishment of cardiac cytoarchitecture in the developing mouse heart. Dev. Biol. 289, 430–441 (2006)

  41. 41.

    et al. Opposing effects of fructokinase C and A isoforms on fructose-induced metabolic syndrome in mice. Proc. Natl Acad. Sci. USA 109, 4320–4325 (2012)

  42. 42.

    , , & Ultrahigh performance liquid chromatography-tandem mass spectrometry method for fast and robust quantification of anionic and aromatic metabolites. Anal. Chem. 82, 4403–4412 (2010)

  43. 43.

    , , & High-throughput, accurate mass metabolome profiling of cellular extracts by flow injection-time-of-flight mass spectrometry. Anal. Chem. 83, 7074–7080 (2011)

  44. 44.

    et al. Cardiotrophin-1 increases angiotensinogen mRNA in rat cardiac myocytes through STAT3: an autocrine loop for hypertrophy. Hypertension 35, 1191–1196 (2000)

  45. 45.

    et al. Cardiomyocyte aldose reductase causes heart failure and impairs recovery from ischemia. PLoS ONE 7, e46549 (2012)

  46. 46.

    Extraction of tissue lipids with a solvent of low toxicity. Methods Enzymol. 72, 5–7 (1981)

  47. 47.

    , & Optimisation of oil red O staining permits combination with immunofluorescence and automated quantification of lipids. Histochem. Cell Biol. 116, 63–68 (2001)

  48. 48.

    et al. Parkin is a lipid-responsive regulator of fat uptake in mice and mutant human cells. J. Clin. Invest. 121, 3701–3712 (2011)

Download references

Acknowledgements

We thank S. Georgiev, T. Simka, S. Xu, C. Bischoff, J. M. Dominguez, W. Kovacs and M. Piontek and other members of the Krek laboratory for discussions, help and technical assistance. We are grateful to M. Stoffel for performing tail vein injections. K. Chien, A. Asipu and R. J. Johnson provided mouse lines. This work was supported by grants from Sinergia (Swiss National Science Foundation) to W.K., T.P. and J. U. and the Swiss Heart Foundation to W.K.

Author information

Author notes

    • Jaya Krishnan
    • , Fiona Grimm
    • , Melis Kayikci
    •  & Jernej Ule

    Present addresses: MRC Clinical Sciences Centre London, Imperial College London, Hammersmith Hospital, Du Cane Road, London W12 ONN, UK (J.K.); MRC National Institute for Medical Research, The Ridgeway, London NW7 1AA, UK (F.G.); MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge Biomedical Campus, Cambridge CB2 0QH, UK (M.K.); Department of Molecular Neuroscience, UCL Institute of Neurology, Queen Square, London WC1N 3BG, UK (J.U.).

    • Peter Mirtschink
    •  & Jaya Krishnan

    These authors contributed equally to this work.

Affiliations

  1. Institute of Molecular Health Sciences, ETH Zurich, 8093 Zürich, Switzerland

    • Peter Mirtschink
    • , Jaya Krishnan
    • , Fiona Grimm
    • , Niklaus Fankhauser
    • , Yann Christinat
    • , Cédric Cortijo
    • , Owen Feehan
    • , Ana Vukolic
    •  & Wilhelm Krek
  2. Department of Medicine, University of Lausanne, 1011 Lausanne, Switzerland

    • Alexandre Sarre
    •  & Thierry Pedrazzini
  3. Institute of Molecular Systems Biology, ETH Zurich, 8093 Zürich, Switzerland

    • Manuel Hörl
    •  & Nicola Zamboni
  4. MRC-Laboratory of Molecular Biology, Hills Road, Cambridge CB2 0QH, UK

    • Melis Kayikci
    •  & Jernej Ule
  5. Universitätsmedizin Göttingen, Klinik für Kardiologie und Pneumologie, D-37075 Göttingen, and DZHK (German Centre for Cardiovascular Research), Partner Site Göttingen, Germany

    • Samuel Sossalla
  6. Department of Anesthesiology and Critical Care Medicine, University Hospital Jena, 07747 Jena, Germany

    • Sebastian N. Stehr

Authors

  1. Search for Peter Mirtschink in:

  2. Search for Jaya Krishnan in:

  3. Search for Fiona Grimm in:

  4. Search for Alexandre Sarre in:

  5. Search for Manuel Hörl in:

  6. Search for Melis Kayikci in:

  7. Search for Niklaus Fankhauser in:

  8. Search for Yann Christinat in:

  9. Search for Cédric Cortijo in:

  10. Search for Owen Feehan in:

  11. Search for Ana Vukolic in:

  12. Search for Samuel Sossalla in:

  13. Search for Sebastian N. Stehr in:

  14. Search for Jernej Ule in:

  15. Search for Nicola Zamboni in:

  16. Search for Thierry Pedrazzini in:

  17. Search for Wilhelm Krek in:

Contributions

P.M, J.K. and W.K. designed and P.M. executed most experiments. F.G. performed ketohexokinase assays, ex vivo glucose/fructose uptake and lipid loading studies. A.S. performed mouse surgeries, echocardiography and necropsy analysis under the supervision of T.P. M.H. and N.Z. performed metabolomic analysis. J.U. and M.K. performed splice junction microarrays and initial data analysis. C.C. performed ATP measurements and Sf3b1-rescue experiments, O.F. quantified lipids and A.V. performed biodistribution experiments. N.F. and Y.C. generated the splice factor list and analysed splice junction microarray data. S.S. and S.N.S. provided human left ventricular biopsies. P.M. and W.K, with help from J. K., wrote the paper.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Wilhelm Krek.

Extended data

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    This file contains Supplementary Figures 1-7, Supplementary Table 4, Supplementary Data 1 and Supplementary Western Blots and gels.

Excel files

  1. 1.

    Supplementary Information

    This file contains Supplementary Table 1.

  2. 2.

    Supplementary Information

    This file contains Supplementary Table 2.

  3. 3.

    Supplementary Information

    This file contains Supplementary Table 3.

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/nature14508

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.