title: Multi-kinase inhibitor C1 triggers mitotic catastrophe of glioma stem cells mainly through MELK kinase inhibition.
abstract: Glioblastoma multiforme (GBM) is a highly lethal brain tumor. Due to resistance to current therapies, patient prognosis remains poor and development of novel and effective GBM therapy is crucial. Glioma stem cells (GSCs) have gained attention as a therapeutic target in GBM due to their relative resistance to current therapies and potent tumor-initiating ability. Previously, we identified that the mitotic kinase maternal embryonic leucine-zipper kinase (MELK) is highly expressed in GBM tissues, specifically in GSCs, and its expression is inversely correlated with the post-surgical survival period of GBM patients. In addition, patient-derived GSCs depend on MELK for their survival and growth both in vitro and in vivo. Here, we demonstrate evidence that the role of MELK in the GSC survival is specifically dependent on its kinase activity. With in silico structure-based analysis for protein-compound interaction, we identified the small molecule Compound 1 (C1) is predicted to bind to the kinase-active site of MELK protein. Elimination of MELK kinase activity was confirmed by in vitro kinase assay in nano-molar concentrations. When patient-derived GSCs were treated with C1, they underwent mitotic arrest and subsequent cellular apoptosis in vitro, a phenotype identical to that observed with shRNA-mediated MELK knockdown. In addition, C1 treatment strongly induced tumor cell apoptosis in slice cultures of GBM surgical specimens and attenuated growth of mouse intracranial tumors derived from GSCs in a dose-dependent manner. Lastly, C1 treatment sensitizes GSCs to radiation treatment. Collectively, these data indicate that targeting MELK kinase activity is a promising approach to attenuate GBM growth by eliminating GSCs in tumors.
authors.affiliation: Department of Neurological Surgery, The Ohio State University Medical Center, Columbus, Ohio, United States of America.
authors.forename: Mutsuko
authors.initials: M
authors.lastname: Minata
authors.affiliation: Department of Neurological Surgery, The Ohio State University Medical Center, Columbus, Ohio, United States of America; Departments of Neurosurgery, Beijing Sanbo Brain Hospital, Capital Medical University, Beijing, China.
authors.forename: Chunyu
authors.initials: C
authors.lastname: Gu
authors.affiliation: Department of Neurological Surgery, The Ohio State University Medical Center, Columbus, Ohio, United States of America.
authors.forename: Kaushal
authors.initials: K
authors.lastname: Joshi
authors.affiliation: Department of Internal Medicine, The Ohio State University Medical Center, Columbus, Ohio, United States of America.
authors.forename: Mariko
authors.initials: M
authors.lastname: Nakano-Okuno
authors.affiliation: Department of Neurological Surgery, The Ohio State University Medical Center, Columbus, Ohio, United States of America.
authors.forename: Christopher
authors.initials: C
authors.lastname: Hong
authors.affiliation: Pharmaco-chemistry, UMR 176 CNRS-Institut Curie, Orsay, France.
authors.forename: Chi-Hung
authors.initials: CH
authors.lastname: Nguyen
authors.affiliation: Departments of Psychiatry and Molecular and Medical Pharmacology, University of California Los Angeles, Los Angeles, California, United States of America.
authors.forename: Harley I
authors.initials: HI
authors.lastname: Kornblum
authors.affiliation: Institut Albert Bonniot, Université Joseph Fourier, Grenoble Cedex 9, France.
authors.forename: Annie
authors.initials: A
authors.lastname: Molla
authors.affiliation: Department of Neurological Surgery, The Ohio State University Medical Center, Columbus, Ohio, United States of America; James Comprehensive Cancer Center, The Ohio State University Medical Center, Columbus, Ohio, United States of America.
authors.forename: Ichiro
authors.initials: I
authors.lastname: Nakano
mesh_terms: D000818:Animals; D002470:Cell Survival; D050718:Complement C1 Inhibitor Protein / Q000494:pharmacology*; D004249:DNA Damage; D004260:DNA Repair; D015536:Down-Regulation; D005910:Glioma / Q000378:metabolism*; D064593:Heterografts / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D016867:Immunocompromised Host; D007150:Immunohistochemistry; D051379:Mice; D008958:Models, Molecular; D014411:Neoplastic Stem Cells / Q000378:metabolism*; D010455:Peptides / Q000494:pharmacology; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D014407:Tumor Cells, Cultured
chemical_list: D050718:Complement C1 Inhibitor Protein; D010455:Peptides; D047428:Protein Kinase Inhibitors; C024711:siomycin A; C484610:MELK protein, human; D017346:Protein Serine-Threonine Kinases
references.citation: Cancer Res. 2008 Sep 15;68(18):7270-7
references.pmid: 18794113
references.citation: Mol Cancer Ther. 2011 Oct;10(10):1818-28
references.pmid: 21859839
references.citation: J Clin Invest. 2008 Sep;118(9):3109-22
references.pmid: 18677408
references.citation: J Biol Chem. 2008 Sep 5;283(36):24848-59
references.pmid: 18611854
references.citation: Sci Signal. 2010 Nov 09;3(147):ra81
references.pmid: 21062993
references.citation: Proc Natl Acad Sci U S A. 2013 May 21;110(21):8644-9
references.pmid: 23650391
references.citation: Cell Cycle. 2009 Nov 1;8(21):3608-9
references.pmid: 19838060
references.citation: Mol Cancer Res. 2009 Mar;7(3):383-92
references.pmid: 19276180
references.citation: Breast Cancer Res. 2007;9(1):R17
references.pmid: 17280616
references.citation: Nat Rev Mol Cell Biol. 2011 Jun;12(6):385-92
references.pmid: 21527953
references.citation: Cancer Res. 2010 Nov 1;70(21):8863-73
references.pmid: 20861186
references.citation: J Neurosci Res. 2008 Jan;86(1):48-60
references.pmid: 17722061
references.citation: FASEB J. 1995 May;9(8):576-96
references.pmid: 7768349
references.citation: Nature. 2006 Dec 7;444(7120):756-60
references.pmid: 17051156
references.citation: Int J Radiat Biol. 1997 Oct;72(4):409-21
references.pmid: 9343106
references.citation: J Cell Biol. 2005 Aug 1;170(3):413-27
references.pmid: 16061694
references.citation: Stem Cells. 2013 May;31(5):870-81
references.pmid: 23339114
references.citation: Cell Cycle. 2005 Jun;4(6):806-11
references.pmid: 15908796
references.citation: Brain Pathol. 2012 Sep;22(5):677-88
references.pmid: 22257080
references.citation: J Vis Exp. 2011 Jul 29;(53):
references.pmid: 21841761
references.citation: Mol Cancer Res. 2009 Jul;7(7):989-99
references.pmid: 19609002
references.citation: J Chem Inf Model. 2012 May 25;52(5):1345-55
references.pmid: 22540736
references.citation: ChemMedChem. 2013 Feb;8(2):289-96
references.pmid: 23281044
references.citation: Cancer Res. 2008 Jul 15;68(14):5706-15
references.pmid: 18632623
references.citation: Cancer Res. 2010 Mar 1;70(5):2030-40
references.pmid: 20145155
references.citation: Oncogene. 2002 Oct 31;21(50):7630-41
references.pmid: 12400006
references.citation: J Mol Med (Berl). 2013 Feb;91(2):237-48
references.pmid: 22945237
references.citation: Cancer Cell. 2008 Jan;13(1):69-80
references.pmid: 18167341
references.citation: Acta Neuropathol. 2007 Aug;114(2):97-109
references.pmid: 17618441
references.citation: J Neurosurg. 2010 Aug;113(2):310-8
references.pmid: 20205512
references.citation: Biol Cell. 2006 Apr;98(4):253-63
references.pmid: 16159311
references.citation: Curr Probl Cancer. 2008 May-Jun;32(3):124-42
references.pmid: 18501775
references.citation: Clin Cancer Res. 2010 May 15;16(10):2890-8
references.pmid: 20460474
references.citation: Lancet Oncol. 2009 May;10(5):459-66
references.pmid: 19269895
references.citation: Clin Cancer Res. 2012 Mar 1;18(5):1268-80
references.pmid: 22230766
references.citation: Genes Dev. 2007 Nov 1;21(21):2683-710
references.pmid: 17974913
references.citation: Neuro Oncol. 2011 Jun;13(6):622-34
references.pmid: 21558073
references.citation: Radiat Res. 2011 Jul;176(1):49-61
references.pmid: 21361780
references.citation: Cancer Res. 2005 Nov 1;65(21):9751-61
references.pmid: 16266996
references.citation: Cell Cycle. 2009 Mar 1;8(5):765-72
references.pmid: 19221479
references.citation: Proc Natl Acad Sci U S A. 2003 Dec 9;100(25):15178-83
references.pmid: 14645703
references.citation: Cell Cycle. 2012 Mar 15;11(6):1167-73
references.pmid: 22370485
references.citation: J Biol Chem. 2008 Dec 12;283(50):34541-53
references.pmid: 18948261
references.citation: Stem Cell Rev. 2008 Sep;4(3):203-10
references.pmid: 18595010
references.citation: Stem Cells. 2013 Jun;31(6):1051-63
references.pmid: 23404835