title: An in vivo chemical genetic screen identifies phosphodiesterase 4 as a pharmacological target for hedgehog signaling inhibition.
abstract: Hedgehog (Hh) signaling plays an integral role in vertebrate development, and its dysregulation has been accepted widely as a driver of numerous malignancies. While a variety of small molecules target Smoothened (Smo) as a strategy for Hh inhibition, Smo gain-of-function mutations have limited their clinical implementation. Modulation of targets downstream of Smo could define a paradigm for treatment of Hh-dependent cancers. Here, we describe eggmanone, a small molecule identified from a chemical genetic zebrafish screen, which induced an Hh-null phenotype. Eggmanone exerts its Hh-inhibitory effects through selective antagonism of phosphodiesterase 4 (PDE4), leading to protein kinase A activation and subsequent Hh blockade. Our study implicates PDE4 as a target for Hh inhibition, suggests an improved strategy for Hh-dependent cancer therapy, and identifies a unique probe of downstream-of-Smo Hh modulation.
authors.affiliation: Division of Cardiovascular Medicine, Department of Medicine, Vanderbilt University School of Medicine, Nashville, TN 37232, USA; Vanderbilt Institute of Chemical Biology, Vanderbilt University School of Medicine, Nashville, TN 37232, USA; Cell and Developmental Biology, Vanderbilt University School of Medicine, Nashville, TN 37232, USA.
authors.forename: Charles H
authors.initials: CH
authors.lastname: Williams
authors.affiliation: Division of Cardiovascular Medicine, Department of Medicine, Vanderbilt University School of Medicine, Nashville, TN 37232, USA; Vanderbilt Institute of Chemical Biology, Vanderbilt University School of Medicine, Nashville, TN 37232, USA.
authors.forename: Jonathan E
authors.initials: JE
authors.lastname: Hempel
authors.affiliation: Division of Cardiovascular Medicine, Department of Medicine, Vanderbilt University School of Medicine, Nashville, TN 37232, USA; College of Veterinary Medicine, Western University of Health Sciences, Pomona, CA 91766, USA.
authors.forename: Jijun
authors.initials: J
authors.lastname: Hao
authors.affiliation: Division of Cardiovascular Medicine, Department of Medicine, Vanderbilt University School of Medicine, Nashville, TN 37232, USA.
authors.forename: Audrey Y
authors.initials: AY
authors.lastname: Frist
authors.affiliation: Division of Cardiovascular Medicine, Department of Medicine, Vanderbilt University School of Medicine, Nashville, TN 37232, USA.
authors.forename: Michelle M
authors.initials: MM
authors.lastname: Williams
authors.affiliation: Cell and Developmental Biology, Vanderbilt University School of Medicine, Nashville, TN 37232, USA.
authors.forename: Jonathan T
authors.initials: JT
authors.lastname: Fleming
authors.affiliation: Vanderbilt Institute of Chemical Biology, Vanderbilt University School of Medicine, Nashville, TN 37232, USA; Department of Chemistry, Vanderbilt University, Nashville, TN 37235, USA.
authors.forename: Gary A
authors.initials: GA
authors.lastname: Sulikowski
authors.affiliation: Department of Neurology, Vanderbilt University School of Medicine, Nashville, TN 37232, USA; Research Medicine, Veterans Affairs Tennessee Valley Healthcare System, Nashville, TN 37212, USA.
authors.forename: Michael K
authors.initials: MK
authors.lastname: Cooper
authors.affiliation: Cell and Developmental Biology, Vanderbilt University School of Medicine, Nashville, TN 37232, USA.
authors.forename: Chin
authors.initials: C
authors.lastname: Chiang
authors.affiliation: Division of Cardiovascular Medicine, Department of Medicine, Vanderbilt University School of Medicine, Nashville, TN 37232, USA; Vanderbilt Institute of Chemical Biology, Vanderbilt University School of Medicine, Nashville, TN 37232, USA; Cell and Developmental Biology, Vanderbilt University School of Medicine, Nashville, TN 37232, USA; Department of Pharmacology, Vanderbilt University School of Medicine, Nashville, TN 37232, USA; Research Medicine, Veterans Affairs Tennessee Valley Healthcare System, Nashville, TN 37212, USA. Electronic address: charles.c.hong@vanderbilt.edu.
authors.forename: Charles C
authors.initials: CC
authors.lastname: Hong
mesh_terms: D000818:Animals; D017868:Cyclic AMP-Dependent Protein Kinases / Q000096:biosynthesis* / Q000235:genetics; D054703:Cyclic Nucleotide Phosphodiesterases, Type 4 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D053823:Hedgehog Proteins / Q000037:antagonists & inhibitors / Q000235:genetics; D058988:Phosphodiesterase 4 Inhibitors / Q000008:administration & dosage* / Q000737:chemistry / Q000302:isolation & purification; D011744:Pyrimidinones / Q000737:chemistry*; D043562:Receptors, G-Protein-Coupled / Q000037:antagonists & inhibitors / Q000235:genetics*; D015398:Signal Transduction / Q000187:drug effects; D054852:Small Molecule Libraries / Q000008:administration & dosage / Q000737:chemistry; D000072086:Smoothened Receptor; D013876:Thiophenes / Q000737:chemistry*; D015533:Transcriptional Activation / Q000187:drug effects; D015027:Zebrafish; D029961:Zebrafish Proteins / Q000037:antagonists & inhibitors / Q000235:genetics*
chemical_list: D053823:Hedgehog Proteins; D058988:Phosphodiesterase 4 Inhibitors; D011744:Pyrimidinones; D043562:Receptors, G-Protein-Coupled; D054852:Small Molecule Libraries; D000072086:Smoothened Receptor; D013876:Thiophenes; D029961:Zebrafish Proteins; C000603950:eggmanone; C491021:smo protein, zebrafish; D017868:Cyclic AMP-Dependent Protein Kinases; D054703:Cyclic Nucleotide Phosphodiesterases, Type 4
references.citation: Genes Dev. 1999 Nov 1;13(21):2828-37
references.pmid: 10557210
references.citation: Dev Biol. 2009 Feb 1;326(1):177-89
references.pmid: 19056373
references.citation: Nature. 2000 Aug 31;406(6799):1005-9
references.pmid: 10984056
references.citation: Proc Natl Acad Sci U S A. 2002 Oct 29;99(22):14071-6
references.pmid: 12391318
references.citation: Genes Dev. 2002 Nov 1;16(21):2743-8
references.pmid: 12414725
references.citation: J Biol. 2002 Nov 6;1(2):10
references.pmid: 12437772
references.citation: Dev Biol. 2004 Nov 1;275(1):143-57
references.pmid: 15464578
references.citation: Development. 1996 Sep;122(9):2835-46
references.pmid: 8787757
references.citation: Development. 1996 Dec;123:153-64
references.pmid: 9007237
references.citation: Development. 1996 Dec;123:255-62
references.pmid: 9007245
references.citation: Development. 2005 Jul;132(13):3103-11
references.pmid: 15930098
references.citation: Development. 2005 Sep;132(17):3977-88
references.pmid: 16049113
references.citation: Nature. 2005 Oct 13;437(7061):1018-21
references.pmid: 16136078
references.citation: PLoS Genet. 2005 Oct;1(4):e53
references.pmid: 16254602
references.citation: Nat Chem Biol. 2006 Jan;2(1):29-30
references.pmid: 16408088
references.citation: Methods Mol Biol. 2009;486:43-55
references.pmid: 19347615
references.citation: Genes Dev. 2009 Aug 15;23(16):1910-28
references.pmid: 19684112
references.citation: Proc Natl Acad Sci U S A. 2009 Aug 18;106(33):14132-7
references.pmid: 19666565
references.citation: Nat Med. 2009 Sep;15(9):1055-61
references.pmid: 19701205
references.citation: Bioorg Med Chem Lett. 2009 Oct 1;19(19):5576-81
references.pmid: 19716296
references.citation: N Engl J Med. 2009 Sep 17;361(12):1164-72
references.pmid: 19726763
references.citation: Science. 2009 Oct 23;326(5952):572-4
references.pmid: 19726788
references.citation: J Cell Sci. 2010 Jan 1;123(Pt 1):62-9
references.pmid: 20016067
references.citation: Genes Dev. 2010 Apr 1;24(7):670-82
references.pmid: 20360384
references.citation: J Cell Biol. 2010 Oct 18;191(2):415-28
references.pmid: 20956384
references.citation: PLoS One. 2010;5(12):e15900
references.pmid: 21209912
references.citation: J Vis Exp. 2010;(46). pii: 2243. doi: 10.3791/2243
references.pmid: 21248690
references.citation: Trends Pharmacol Sci. 2011 Jun;32(6):337-44
references.pmid: 21450351
references.citation: Nat Rev Cancer. 2011 Jul;11(7):493-501
references.pmid: 21614026
references.citation: Development. 2000 May;127(10):2189-99
references.pmid: 10769242
references.citation: Genes Dev. 2014 Nov 15;28(22):2547-63
references.pmid: 25403183
references.citation: Nat Med. 2014 Sep;20(9):1035-42
references.pmid: 25150496
references.citation: Cell Rep. 2014 Jan 16;6(1):168-81
references.pmid: 24373970
references.citation: Oncogene. 2013 Dec 12;32(50):5574-81
references.pmid: 23995793
references.citation: Cell Signal. 2013 Nov;25(11):2222-30
references.pmid: 23872071
references.citation: BMC Biol. 2013;11:37
references.pmid: 23587183
references.citation: J Cell Biol. 2012 Aug 20;198(4):607-21
references.pmid: 22908311
references.citation: Exp Cell Res. 2012 Oct 1;318(16):1959-72
references.pmid: 22659135
references.citation: J Biol Chem. 2012 May 25;287(22):17905-13
references.pmid: 22474285
references.citation: Annu Rev Cell Dev Biol. 2011;27:513-37
references.pmid: 21801010
references.citation: Development. 2011 Nov;138(22):4921-30
references.pmid: 22007132
references.citation: Nature. 2012 Apr 5;484(7392):125-9
references.pmid: 22425997
references.citation: Mol Cancer Res. 2015 Jan;13(1):149-60
references.pmid: 25149359
references.citation: Clin Cancer Res. 2008 Dec 1;14(23):7717-25
references.pmid: 19047098
references.citation: Science. 2007 Jul 20;317(5836):372-6
references.pmid: 17641202