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A defective response to Hedgehog signaling in disorders of cholesterol biosynthesis

Nature Genetics volume 33, pages 508–513 (2003)Cite this article

A Corrigendum to this article was published on 01 May 2003

Abstract

Smith–Lemli–Opitz syndrome (SLOS), desmosterolosis and lathosterolosis are human syndromes caused by defects in the final stages of cholesterol biosynthesis. Many of the developmental malformations in these syndromes occur in tissues and structures whose embryonic patterning depends on signaling by the Hedgehog (Hh) family of secreted proteins. Here we report that response to the Hh signal is compromised in mutant cells from mouse models of SLOS and lathosterolosis and in normal cells pharmacologically depleted of sterols. We show that decreasing levels of cellular sterols correlate with diminishing responsiveness to the Hh signal. This diminished response occurs at sterol levels sufficient for normal autoprocessing of Hh protein, which requires cholesterol as cofactor and covalent adduct. We further find that sterol depletion affects the activity of Smoothened (Smo), an essential component of the Hh signal transduction apparatus.

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Figure 1: Cholesterol biosynthesis and Hh pathway.
Figure 2: Cyclodextrin treatment inhibited Shh signaling.
Figure 3: Cells defective in cholesterol biosynthesis did not respond to Shh.
Figure 4: Inhibition of Hh signal response by sterol depletion in cells with intact cholesterol biosynthesis.
Figure 5: Sterol depletion inhibited Shh signaling downstream of Ptch at the level of Smo.

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References

  1. Porter, J.A., Young, K.E. & Beachy, P.A. Cholesterol modification of hedgehog signaling proteins in animal development. Science 274, 255–259 (1996).

    Article  CAS  Google Scholar 

  2. Beachy, P.A. et al. Multiple roles of cholesterol in hedgehog protein biogenesis and signaling. Cold Spring Harb. Symp. Quant. Biol. 62, 191–204 (1997).

    Article  CAS  Google Scholar 

  3. Cooper, M.K., Porter, J.A., Young, K.E. & Beachy, P.A. Teratogen-mediated inhibition of target tissue response to Shh signaling. Science 280, 1603–1607 (1998).

    Article  CAS  Google Scholar 

  4. Chen, J.K., Taipale, J., Cooper, M.K. & Beachy, P.A. Inhibition of Hedgehog signaling by direct binding of cyclopamine to Smoothened. Genes Dev. 16, 2743–2748 (2002).

    Article  CAS  Google Scholar 

  5. Incardona, J.P. et al. Cyclopamine inhibition of Sonic hedgehog signal transduction is not mediated through effects on cholesterol transport. Dev. Biol. 224, 440–452 (2000).

    Article  CAS  Google Scholar 

  6. Ohtani, Y., Irie, T., Uekama, K., Fukunaga, K. & Pitha, J. Differential effects of α-, β- and γ-cyclodextrins on human erythrocytes. Eur. J. Biochem. 186, 17–22 (1989).

    Article  CAS  Google Scholar 

  7. Kilsdonk, E.P. et al. Cellular cholesterol efflux mediated by cyclodextrins. J. Biol. Chem. 270, 17250–17256 (1995).

    Article  CAS  Google Scholar 

  8. Chiang, C. et al. Cyclopia and defective axial patterning in mice lacking Sonic hedgehog gene function. Nature 383, 407–413 (1996).

    Article  CAS  Google Scholar 

  9. Liem, K.F. Jr., Tremml, G., Roelink, H. & Jessell, T.M. Dorsal differentiation of neural plate cells induced by BMP-mediated signals from epidermal ectoderm. Cell 82, 969–979 (1995).

    Article  CAS  Google Scholar 

  10. Guy, R.K. Inhibition of sonic hedgehog autoprocessing in cultured mammalian cells by sterol deprivation. Proc. Natl. Acad. Sci. USA 97, 7307–7312 (2000).

    Article  CAS  Google Scholar 

  11. Wassif, C.A. et al. Mutations in the human sterol δ7-reductase gene at 11q12–13 cause Smith–Lemli–Opitz syndrome. Am. J. Hum. Genet. 63, 55–62 (1998).

    Article  CAS  Google Scholar 

  12. Kelley, R.I. Inborn errors of cholesterol biosynthesis. Adv. Pediatr. 47, 1–53 (2000).

    CAS  PubMed  Google Scholar 

  13. Brown, M.S., Faust, J.R. & Goldstein, J.L. Induction of 3-hydroxy-3-methylglutaryl coenzyme A reductase activity in human fibroblasts incubated with compactin (ML-236B), a competitive inhibitor of the reductase. J. Biol. Chem. 253, 1121–1128 (1978).

    CAS  PubMed  Google Scholar 

  14. Bittman, R. & Fischkoff, S.A. Fluorescence studies of the binding of the polyene antibiotics filipin 3, amphotericin B, nystatin, and lagosin to cholesterol. Proc. Natl. Acad. Sci. USA 69, 3795–3799 (1972).

    Article  CAS  Google Scholar 

  15. Goodrich, L.V., Milenkovic, L., Higgins, K.M. & Scott, M.P. Altered neural cell fates and medulloblastoma in mouse patched mutants. Science 277, 1109–1113 (1997).

    Article  CAS  Google Scholar 

  16. Taipale, J. et al. Effects of oncogenic mutations in Smoothened and Patched can be reversed by cyclopamine. Nature 406, 1005–1009 (2000).

    Article  CAS  Google Scholar 

  17. Gimpl, G., Burger, K. & Fahrenholz, F. Cholesterol as modulator of receptor function. Biochemistry 36, 10959–10974 (1997).

    Article  CAS  Google Scholar 

  18. Simons, K. & Ikonen, E. Functional rafts in cell membranes. Nature 387, 569–572 (1997).

    Article  CAS  Google Scholar 

  19. Nowaczyk, M.J., McCaughey, D., Whelan, D.T. & Porter, F.D. Incidence of Smith–Lemli–Opitz syndrome in Ontario, Canada. Am. J. Med. Genet. 102, 18–20 (2001).

    Article  CAS  Google Scholar 

  20. Matsunaga, E. & Shiota, K. Holoprosencephaly in human embryos: epidemiologic studies of 150 cases. Teratology 16, 261–272 (1977).

    Article  CAS  Google Scholar 

  21. Roelink, H. et al. Floor plate and motor neuron induction by different concentrations of the amino-terminal cleavage product of sonic hedgehog autoproteolysis. Cell 81, 445–455 (1995).

    Article  CAS  Google Scholar 

  22. Sasaki, H., Hui, C., Nakafuku, M. & Kondoh, H. A binding site for Gli proteins is essential for HNF-3β floor plate enhancer activity in transgenics and can respond to Shh in vitro. Development 124, 1313–1322 (1997).

    CAS  PubMed  Google Scholar 

  23. Gibson, K.M. et al. 3-Hydroxy-3-methylglutaryl coenzyme A reductase activity in cultured fibroblasts from patients with mevalonate kinase deficiency: differential response to lipid supplied by fetal bovine serum in tissue culture medium. J. Lipid Res. 31, 515–521 (1990).

    CAS  PubMed  Google Scholar 

  24. Kelley, R.I. Diagnosis of Smith–Lemli–Opitz syndrome by gas chromatography/mass spectrometry of 7-dehydrocholesterol in plasma, amniotic fluid and cultured skin fibroblasts. Clin. Chim. Acta 236, 45–58 (1995).

    Article  CAS  Google Scholar 

  25. Donnai, D., Burn, J. & Hughes, H. Smith–Lemli–Opitz syndromes: do they include the Pallister–Hall syndrome? Am. J. Med. Genet. 28, 741–743 (1987).

    Article  CAS  Google Scholar 

  26. Parnes, S., Hunter, A.G., Jimenez, C., Carpenter, B.F. & MacDonald, I. Apparent Smith–Lemli–Opitz syndrome in a child with a previously undescribed form of mucolipidosis not involving the neurons. Am. J. Med. Genet. 35, 397–405 (1990).

    Article  CAS  Google Scholar 

  27. Clayton, P., Mills, K., Keeling, J. & FitzPatrick, D. Desmosterolosis: a new inborn error of cholesterol biosynthesis. Lancet 348, 404 (1996).

    Article  CAS  Google Scholar 

  28. Ingham, P.W. Hedgehog signaling: a tale of two lipids. Science 294, 1879–1881 (2001).

    Article  CAS  Google Scholar 

  29. Wang, B., Fallon, J.F. & Beachy, P.A. Hedgehog-regulated processing of Gli3 produces an anterior/posterior repressor gradient in the developing vertebrate limb. Cell 100, 423–434 (2000).

    Article  CAS  Google Scholar 

  30. Chamoun, Z. et al. Skinny hedgehog, an acyltransferase required for palmitoylation and activity of the hedgehog signal. Science 293, 2080–2084 (2001).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank R.K. Mann and D. Valle for critical review of this manuscript. M.K.C was supported by a career development award from the Burroughs Wellcome Fund and a US National Institutes of Health K08 award from the National Institute of Neurological Disorders and Stroke. This work was supported in part by a grant from the US National Institutes of Health (P.A.B.). P.A.B. is an investigator of the Howard Hughes Medical Institute.

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Author notes

  1. Michael K. Cooper

    Present address: Department of Neurology, Vanderbilt University Medical Center, Nashville, Tennessee, 37232, USA

Authors and Affiliations

  1. Department of Molecular Biology and Genetics, Howard Hughes Medical Institute, Johns Hopkins University School of Medicine, Baltimore, 21205, Maryland, USA

    Michael K. Cooper, Jussi Taipale, Ruoyu Gong & Philip A. Beachy

  2. Department of Neurology, Howard Hughes Medical Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland, 21205, USA

    Michael K. Cooper

  3. Heritable Disorders Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, 20892-1830, Maryland, USA

    Christopher A. Wassif, Patrycja A. Krakowiak & Forbes D. Porter

  4. Kennedy Krieger Institute, Baltimore, 21205, Maryland, USA

    Richard I. Kelley

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Correspondence to Philip A. Beachy.

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Cooper, M., Wassif, C., Krakowiak, P. et al. A defective response to Hedgehog signaling in disorders of cholesterol biosynthesis. Nat Genet 33, 508–513 (2003). https://doi.org/10.1038/ng1134

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