DOCSLIB.ORG

Chromosomal)Instability)In)Glioma)Stem)Cell) Lines)From)Glioblastoma)Multiforme:)) Implications)For)New)Therapeutic)Strategies) ' ' ' ' ' '

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Chromosomal)Instability)In)Glioma)Stem)Cell) Lines)From)Glioblastoma)Multiforme:)) Implications)For)New)Therapeutic)Strategies) ' ' ' ' ' ' SCUOLA'DI'DOTTORATO' UNIVERSITÀ'DEGLI'STUDI'DI'MILANO2BICOCCA' ' ' School'of'Medicine'and'Surgery' ' PhD'program'in'Neuroscience'' Cycle'XXIX' ' ' ' ' ' ' ' CHROMOSOMAL)INSTABILITY)IN)GLIOMA)STEM)CELL) LINES)FROM)GLIOBLASTOMA)MULTIFORME:)) IMPLICATIONS)FOR)NEW)THERAPEUTIC)STRATEGIES) ' ' ' ' ' ' Surname:'CILIBRASI''''Name:'CHIARA' Registration'number:'709467' ' ' ' Tutor:'Dr.'ANGELA'BENTIVEGNA' Coordinator:'Prof.'GUIDO'ANGELO'CAVALETTI' ' ' ' ' ' Academic'Year'2015/2016 TABLE)OF)CONTENT) Summary)...............................................................................................................)IV)) Abbreviations)........................................................................................................)VI)) Figures)...................................................................................................................)IX)) Tables)....................................................................................................................)XI) ' CHapter)1:)Introduction).........................................................................................)1) ' 1.1'Gliomas'..............................................................................................................'2' 1.2'Glioblastoma'multiforme'....................................................................................'5' 1.2.1'Glioblastoma'standard'of'care'.................................................................'12' 1.3'Cancer'stem'cells'theory'.................................................................................'16' 1.3.1'Glioma'stem'cells'.....................................................................................'18' 1.3.1.1'Glioma'stem'cells'markers'................................................................'19' 1.3.1.2'Glioma'stem'cells'implicated'signalling'pathways'............................'22' 1.3.1.3'Glioma'stem'cells'microenvironment'................................................'24' 1.3.1.4'Glioma'stem'cells2dependent'therapeutic'resistance'.......................'28' 1.4'Symmetric'and'asymmetric'cell'division'..........................................................'31' 1.4.1'Symmetric'and'asymmetric'cell'division'in'D.Melanogaster'stem2like'cells' '..........................................................................................................................'31' 1.4.2'Cell'division'mode'in'mammalian'neural'stem'cells''................................'35' 1.4.3'Evidence'of'GSCs'division'mode''............................................................'37' 1.5'Chromosomal'instability'(CIN)'.........................................................................'40' 1.5.1'Methods'for'the'analysis'of'CIN'...............................................................'41' 1.5.2'Causes'of'CIN'..........................................................................................'43' 1.5.3'CIN'and'tumorigenesis'.............................................................................'50' 1.5.3.1'Aurora'kinases'and'cancer'...............................................................'51' 1.6'CIN'as'a'therapeutic'target'..............................................................................'57' 1.6.1'Aurora'kinases'inhibitors'..........................................................................'58' 1.6.1.1'Danusertib'.........................................................................................'59' ' CHapter)2:)Aim)of)the)work).................................................................................)66) ' CHapter)3:)Effect)of)Aurora)kinases)inHibition)on)GSCs).................................)68) Materials)and)Methods)........................................................................................)69) ' 3.1'Materials'..........................................................................................................'70' 3.1.1'Cell'lines'...................................................................................................'70' 3.1.2'Drugs'........................................................................................................'70' 3.1.3'Solutions'and'Buffers'...............................................................................'71' 3.1.4'Other'Reagents'........................................................................................'71' 3.2'Methods'...........................................................................................................'75' 3.2.1'Cell'culture'conditions'..............................................................................'75' 3.2.2'RNA'extraction'.........................................................................................'75' 3.2.3'TaqMan'gene'expression'assay'..............................................................'75' I' 3.2.4'MTT'assay'................................................................................................'76' 3.2.5'Trypan'blue'dye'exclusion'assay'.............................................................'76' 3.2.6'Conventional'cytogenetics'.......................................................................'77' 3.2.7'Fluorescence'in.situ'hybridization'............................................................'77' 3.2.8'Neurosphere'formation'assay'..................................................................'78' 3.2.9'Cell'morphology'analysis'.........................................................................'78' 3.2.10'Giemsa'staining'......................................................................................'78' 3.2.11'Western'Blot'...........................................................................................'79' 3.2.12'Cell'cycle'profile'analysis'.......................................................................'79' 3.2.13'Immunofluorescence'..............................................................................'80' 3.2.14'Live'cell'imaging'.....................................................................................'80' 3.2.15'DNA'extraction'.......................................................................................'81' 3.2.16'TP53'Sanger'sequencing'.......................................................................'81' 3.2.17'Microsatellites'analysis'...........................................................................'82' 3.2.18'DNA'integrity'evaluation'.........................................................................'82' 3.2.19'β'galactosidase'staining'.........................................................................'83' 3.2.20''Statistics'................................................................................................'83' Results)..................................................................................................................)84) ' 3.3'Preliminary'results:'GSCs'are'characterized'by'enhanced'CIN'and'AURK' CNAs'.....................................................................................................................'85' 3.4'Aurora'kinases'A'and'B'are'overexpressed'in'GSC'lines'...............................'86' 3.5'Aurora'kinases'protein'levels'...........................................................................'87' 3.6'Sensitivity'and'resistance'to'Danusertib'measured'by'metabolic'activity'........'88' 3.7'Danusertib'effect'on'GSCs'viability'.................................................................'90' 3.8'Danusertib'induces'strong'changes'in'cell'and'nuclei'morphology'in'sensitive' GSCs'.....................................................................................................................'91' 3.9'Danusertib'induces'ploidy'increase'in'GSC'lines'............................................'93' 3.10'Danusertib'effect'on'GSCs'proliferation'........................................................'95' 3.11'Danusertib'effect'on'GSCs'clonogenic'potential'...........................................'96' 3.12'Danusertib'induces'a'decrease'in'phospho2Aurora'kinases'levels'in'GSC' lines'.......................................................................................................................'97' 3.13'Live'imaging'analysis'of'GSCs'fate'after'Danusertib'exposure'.....................'98' 3.14'Different'sensitivity'of'GSCs'to'Aurora'inhibition'is'not'dependent'on'TP53' status'...................................................................................................................'100' 3.15'TP53'loss'of'heterozygosity'(LOH)'analysis'................................................'102' 3.16'Sensitive'and'resistant'GSCs'have'a'different'cell'cycle'distribution'..........'103' 3.17'Danusertib'does'not'induce'any'DNA'fragmentation'...................................'106' 3.18'Aurora'inhibition'induces'an'increase'in'senescence'in'sensitive'GSCs'....'106' 3.19'Multiple'rounds'of'Aurora'inhibition'induce'an'increase'in'ploidy'and' senescence'also'in'resistant'GSCs'.....................................................................'107' ' CHapter)4:)CHaracterization)of)GSCs)division)mode).....................................)110) Materials)and)Methods)......................................................................................)111) ' 4.1'Materials'........................................................................................................'112' 4.1.1'Cell'lines'.................................................................................................'112' 4.1.2'Solutions'and'Buffers'.............................................................................'112'
Load more

Recommended publications
  • Aurora Kinase a in Gastrointestinal Cancers: Time to Target Ahmed Katsha1, Abbes Belkhiri1, Laura Goff3 and Wael El-Rifai1,2,4*
    Katsha et al. Molecular Cancer (2015) 14:106 DOI 10.1186/s12943-015-0375-4 REVIEW Open Access Aurora kinase A in gastrointestinal cancers: time to target Ahmed Katsha1, Abbes Belkhiri1, Laura Goff3 and Wael El-Rifai1,2,4* Abstract Gastrointestinal (GI) cancers are a major cause of cancer-related deaths. During the last two decades, several studies have shown amplification and overexpression of Aurora kinase A (AURKA) in several GI malignancies. These studies demonstrated that AURKA not only plays a role in regulating cell cycle and mitosis, but also regulates a number of key oncogenic signaling pathways. Although AURKA inhibitors have moved to phase III clinical trials in lymphomas, there has been slower progress in GI cancers and solid tumors. Ongoing clinical trials testing AURKA inhibitors as a single agent or in combination with conventional chemotherapies are expected to provide important clinical information for targeting AURKA in GI cancers. It is, therefore, imperative to consider investigations of molecular determinants of response and resistance to this class of inhibitors. This will improve evaluation of the efficacy of these drugs and establish biomarker based strategies for enrollment into clinical trials, which hold the future direction for personalized cancer therapy. In this review, we will discuss the available data on AURKA in GI cancers. We will also summarize the major AURKA inhibitors that have been developed and tested in pre-clinical and clinical settings. Keywords: Aurora kinases, Therapy, AURKA inhibitors, MNL8237, Alisertib, Gastrointestinal, Cancer, Signaling pathways Introduction stage [9-11]. Furthermore, AURKA is critical for Mitotic kinases are the main proteins that coordinate ac- bipolar-spindle assembly where it interacts with Ran- curate mitotic processing [1].
    [Show full text]
  • Structure-Based Discovery and Bioactivity Evaluation of Novel
    molecules Article Structure-Based Discovery and Bioactivity Evaluation of Novel Aurora-A Kinase Inhibitors as Anticancer Agents via Docking-Based Comparative Intermolecular Contacts Analysis (dbCICA) Majd S. Hijjawi 1 , Reem Fawaz Abutayeh 2 and Mutasem O. Taha 3,* 1 Department of Pharmacology, Faculty of Medicine, The University of Jordan, Amman 11942, Jordan; [email protected] 2 Department of Pharmaceutical Chemistry and Pharmacognosy, Faculty of Pharmacy, Applied Science Private University, Amman 11931, Jordan; [email protected] 3 Department of Pharmaceutical Sciences, Faculty of Pharmacy, University of Jordan, Amman 11942, Jordan * Correspondence: [email protected]; Tel.: +962-6-535-5000 Academic Editors: Helen Osborn, Mohammad Najlah, Jean Jacques Vanden Eynde, Annie Mayence and Tien L. Huang Received: 15 October 2020; Accepted: 11 December 2020; Published: 18 December 2020 Abstract: Aurora-A kinase plays a central role in mitosis, where aberrant activation contributes to cancer by promoting cell cycle progression, genomic instability, epithelial-mesenchymal transition, and cancer stemness. Aurora-A kinase inhibitors have shown encouraging results in clinical trials but have not gained Food and Drug Administration (FDA) approval. An innovative computational workflow named Docking-based Comparative Intermolecular Contacts Analysis (dbCICA) was applied—aiming to identify novel Aurora-A kinase inhibitors—using seventy-nine reported Aurora-A kinase inhibitors to specify the best possible docking settings needed to fit into the active-site binding pocket of Aurora-A kinase crystal structure, in a process that only potent ligands contact critical binding-site spots, distinct from those occupied by less-active ligands. Optimal dbCICA models were transformed into two corresponding pharmacophores.
    [Show full text]
  • Spatiotemporal Dynamics of Aurora B-PLK1-MCAK Signaling Axis
    www.nature.com/scientificreports OPEN Spatiotemporal dynamics of Aurora B-PLK1-MCAK signaling axis orchestrates kinetochore Received: 21 October 2014 Accepted: 13 May 2015 bi-orientation and faithful Published: 24 July 2015 chromosome segregation Hengyi Shao1, Yuejia Huang2,3, Liangyu Zhang1,3, Kai Yuan1, Youjun Chu1,3, Zhen Dou1,2, Changjiang Jin1, Minerva Garcia-Barrio3, Xing Liu1,2,3 & Xuebiao Yao1 Chromosome segregation in mitosis is orchestrated by the dynamic interactions between the kinetochore and spindle microtubules. The microtubule depolymerase mitotic centromere-associated kinesin (MCAK) is a key regulator for an accurate kinetochore-microtubule attachment. However, the regulatory mechanism underlying precise MCAK depolymerase activity control during mitosis remains elusive. Here, we describe a novel pathway involving an Aurora B-PLK1 axis for regulation of MCAK activity in mitosis. Aurora B phosphorylates PLK1 on Thr210 to activate its kinase activity at the kinetochores during mitosis. Aurora B-orchestrated PLK1 kinase activity was examined in real- time mitosis using a fluorescence resonance energy transfer-based reporter and quantitative analysis of native PLK1 substrate phosphorylation. Active PLK1, in turn, phosphorylates MCAK at Ser715 which promotes its microtubule depolymerase activity essential for faithful chromosome segregation. Importantly, inhibition of PLK1 kinase activity or expression of a non-phosphorylatable MCAK mutant prevents correct kinetochore-microtubule attachment, resulting in abnormal anaphase with chromosome bridges. We reason that the Aurora B-PLK1 signaling at the kinetochore orchestrates MCAK activity, which is essential for timely correction of aberrant kinetochore attachment to ensure accurate chromosome segregation during mitosis. During cell division, accurate chromosome segregation requires dynamic interactions between kineto- chores and spindle microtubules (MTs), which results in accurate chromosome bi-orientation1–4.
    [Show full text]
  • The Aurora B Kinase Activity Is Required for the Maintenance of the Differentiated State of Murine Myoblasts
    Cell Death and Differentiation (2009) 16, 321–330 & 2009 Macmillan Publishers Limited All rights reserved 1350-9047/09 $32.00 www.nature.com/cdd The Aurora B kinase activity is required for the maintenance of the differentiated state of murine myoblasts G Amabile1,2, AM D’Alise1, M Iovino1, P Jones3, S Santaguida4, A Musacchio4, S Taylor5 and R Cortese*,1 Reversine is a synthetic molecule capable of inducing dedifferentiation of C2C12, a murine myoblast cell line, into multipotent progenitor cells, which can be redirected to differentiate in nonmuscle cell types under appropriate conditions. Reversine is also a potent inhibitor of Aurora B, a protein kinase required for mitotic chromosome segregation, spindle checkpoint function, cytokinesis and histone H3 phosphorylation, raising the possibility that the dedifferentiation capability of reversine is mediated through the inhibition of Aurora B. Indeed, here we show that several other well-characterized Aurora B inhibitors are capable of dedifferentiating C2C12 myoblasts. Significantly, expressing drug-resistant Aurora B mutants, which are insensitive to reversine block the dedifferentiation process, indicating that Aurora B kinase activity is required to maintain the differentiated state. We show that the inhibition of the spindle checkpoint or cytokinesis per se is not sufficient for dedifferentiation. Rather, our data support a model whereby changes in histone H3 phosphorylation result in chromatin remodeling, which in turn restores the multipotent state. Cell Death and Differentiation (2009) 16, 321–330; doi:10.1038/cdd.2008.156; published online 31 October 2008 Lineage-restricted cells can be reprogramed to a state Evidence is emerging that the role of Aurora B is not of pluripotency by several different manipulations, including restricted to mitosis and cell division.
    [Show full text]
  • Quantitative Conformational Profiling of Kinase Inhibitors Reveals Origins of Selectivity for Aurora Kinase Activation States
    Quantitative conformational profiling of kinase inhibitors reveals origins of selectivity for Aurora kinase activation states Eric W. Lakea, Joseph M. Murettab, Andrew R. Thompsonb, Damien M. Rasmussenb, Abir Majumdara, Erik B. Faberc, Emily F. Ruffa, David D. Thomasb, and Nicholas M. Levinsona,d,1 aDepartment of Pharmacology, University of Minnesota, Minneapolis, MN 55455; bDepartment of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis, MN 55455; cDepartment of Medicinal Chemistry, University of Minnesota, Minneapolis, MN 55455; and dMasonic Cancer Center, University of Minnesota, Minneapolis, MN 55455 Edited by Kevan M. Shokat, University of California, San Francisco, CA, and approved November 7, 2018 (received for review June 28, 2018) Protein kinases undergo large-scale structural changes that tightly lytically important Asp–Phe–Gly (DFG) motif, located on the regulate function and control recognition by small-molecule in- flexible activation loop of the kinase, is flipped relative to its hibitors. Methods for quantifying the conformational effects of orientation in the active state (referred to as “DFG-out,” in inhibitors and linking them to an understanding of selectivity contrast to the active “DFG-in” state). The observation that the patterns have long been elusive. We have developed an ultrafast inactive DFG-out states of kinases are more divergent than the time-resolved fluorescence methodology that tracks structural catalytically competent DFG-in state has led to a focus on type II movements of the kinase activation loop in solution with inhibitors as a potential answer to the selectivity problem (8, 9). angstrom-level precision, and can resolve multiple structural states However, kinome-wide profiling of kinase inhibitors has revealed and quantify conformational shifts between states.
    [Show full text]
  • Functional Interplay Between NF-Κb-Inducing Kinase and C-Abl
    Plasma Cell Disorders ARTICLE Functional interplay between NF- B-inducing kinase and c-Abl kinases limits responseκ to Ferrata Storti Foundation Aurora inhibitors in multiple myeloma Laura Mazzera,1,2 Manuela Abeltino,1 Guerino Lombardi,2 Anna Maria Cantoni,3 Roberto Ria,4 Micaela Ricca,2 Ilaria Saltarella,4 Valeria Naponelli,1 Federica Maria Angela Rizzi,1,5 Roberto Perris,5,6 Attilio Corradi,3 Angelo Vacca,4 Antonio Bonati1,5 and Paolo Lunghi5,6 1 2 Department of Medicine and Surgery, University of Parma, Parma; Istituto Zooprofilattico Haematologica 2019 Sperimentale della Lombardia e dell’Emilia Romagna “Bruno Ubertini,” Brescia; 3Department of Veterinary Science, University of Parma, Parma; 4Department of Volume 104(12):2465-2481 Biomedical Sciences and Human Oncology, Section of Internal Medicine and Clinical Oncology, University of Bari "Aldo Moro" Medical School, Bari; 5Center for Molecular and Translational Oncology, University of Parma, Parma and 6Department of Chemistry, Life Sciences and Environmental Sustainability, University of Parma, Parma, Italy ABSTRACT onsidering that Aurora kinase inhibitors are currently under clinical investigation in hematologic cancers, the identification of molecular Cevents that limit the response to such agents is essential for enhanc- ing clinical outcomes. Here, we discover a NF-κB-inducing kinase (NIK)-c- Abl-STAT3 signaling-centered feedback loop that restrains the efficacy of Aurora inhibitors in multiple myeloma. Mechanistically, we demonstrate that Aurora inhibition promotes NIK protein stabilization via downregula- tion of its negative regulator TRAF2. Accumulated NIK converts c-Abl tyro- sine kinase from a nuclear proapoptotic into a cytoplasmic antiapoptotic effector by inducing its phosphorylation at Thr735, Tyr245 and Tyr412 residues, and, by entering into a trimeric complex formation with c-Abl and Correspondence: STAT3, increases both the transcriptional activity of STAT3 and expression PAOLO LUNGHI of the antiapoptotic STAT3 target genes PIM1 and PIM2.
    [Show full text]
  • BRAF Testing in Advanced Colorectal Cancer: Is It Ready for Prime Time?
    BRAF Testing in Advanced Colorectal Cancer: Is It Ready for Prime Time? Brooke Phillips, MD, Matthew Kalady, MD, and Richard Kim, MD Dr. Phillips is a fellow in the Department Abstract: Given that KRAS mutant colorectal tumors do not respond of Oncology at the Taussig Cancer Institute to anti-EGFR monoclonal antibodies such as cetuximab or panitu- at the Cleveland Clinic, where Dr. Kim is mumab, it is now standard that all patients with metastatic colorectal a Clinical Assistant Professor. Dr. Kalady is cancer who are candidates for these therapies undergo KRAS testing. staff colorectal surgeon in the Department of Colorectal Surgery at the Cleveland BRAF encodes a protein kinase, which is involved in intracellular Clinic, Cleveland, Ohio. signaling and cell growth and is a principal downstream effector of KRAS. BRAF is now increasingly being investigated in metastatic colorectal carcinoma. BRAF mutations occur in less than 10–15% of tumors and appear to be poor prognostic markers. However the predictive nature of this biomarker is yet undefined. This article will Address correspondence to: review the evidence behind both KRAS and BRAF testing in metastatic Richard Kim, MD colorectal cancer. Taussig Cancer Institute Cleveland Clinic, R35 9500 Euclid Ave. Cleveland, OH 44195 Phone: 216-444-0293 Introduction Fax: 216-444-9464 E-mail: [email protected] The advent of target-specific cancer therapeutics has remarkably improved the outcomes of patients with metastatic colorectal cancer (mCRC). The 3 monoclonal antibodies that are approved in treatment of mCRC include cetuximab (Erbitux, ImClone) and panitumumab (Vectibix, Amgen), which are monoclonal antibodies against epidermal growth factor receptor (EGFR), and bevacizumab (Avastin, Genentech), which is a monoclonal antibody against vascular endothelial growth (VEGF) receptor.
    [Show full text]
  • Inhibition of Aurora a and Aurora B Is Required for the Sensitivity of HPV
    Published OnlineFirst May 18, 2017; DOI: 10.1158/1535-7163.MCT-17-0159 Cancer Biology and Signal Transduction Molecular Cancer Therapeutics Inhibition of Aurora A and Aurora B Is Required for the Sensitivity of HPV-Driven Cervical Cancers to Aurora Kinase Inhibitors David Martin1,2, Sora Fallaha3, Martina Proctor1, Alexander Stevenson1, Lewis Perrin4, Nigel McMillan3, and Brian Gabrielli1 Abstract The activity and efficacy of Aurora inhibitors have been the inhibition of Aurora A and B that provides the selectivity and reported in a wide range of cancer types. The most prominent efficacy of this drug in vivo in this disease setting. We also present Aurora inhibitor is alisertib, an investigational Aurora inhibitor formal evidence that alisertib requires progression through mito- that has been the subject of more than 30 clinical trials. Alisertib sis for its efficacy, and that it is unlikely to combine with drugs that has inhibitory activity against both Aurora A and B, although it is promote a G2 DNA damage checkpoint response. This work considered to be primarily an Aurora A inhibitor in vivo. Here, we demonstrates that inhibition of Aurora A and B is required for show that alisertib inhibits both Aurora A and B in vivo in effective control of HPV-driven cancers by Aurora kinase inhibi- preclinical models of HPV-driven cervical cancer, and that it is tors. Mol Cancer Ther; 16(9); 1–8. Ó2017 AACR. Introduction alisertib displayed selectivity for the HPV-dependent cervical cancers in vitro and in xenograft models (9). Alisertib has been Aurora kinases are critical regulators of entry into and exit from reported to be primarily an Aurora A inhibitor, although it mitosis.
    [Show full text]
  • Dual PDK1/Aurora Kinase a Inhibitors Reduce Pancreatic Cancer Cell Proliferation and Colony Formation
    cancers Article Dual PDK1/Aurora Kinase A Inhibitors Reduce Pancreatic Cancer Cell Proliferation and Colony Formation 1, 1, 2 3 3 Ilaria Casari y, Alice Domenichini y, Simona Sestito , Emily Capone , Gianluca Sala , Simona Rapposelli 2 and Marco Falasca 1,* 1 Metabolic Signalling Group, School of Pharmacy and Biomedical Sciences, Curtin Health Innovation Research Institute, Curtin University, Bentley 6102, Australia; [email protected] (I.C.); [email protected] (A.D.) 2 Department of Pharmacy, University of Pisa, Via Bonanno, 6, 56126 Pisa, Italy; [email protected] (S.S.); [email protected] (S.R.) 3 Dipartimento di Scienze Mediche, Orali e Biotecnologiche, University “G. d’Annunzio” di Chieti-Pescara, Center for Advanced Studies and Technology (CAST), 66100 Chieti, Italy; [email protected] (E.C.); [email protected] (G.S.) * Correspondence: [email protected]; Tel.: +61-8-92669712 These authors contributed equally to this work. y Received: 8 October 2019; Accepted: 28 October 2019; Published: 31 October 2019 Abstract: Deregulation of different intracellular signaling pathways is a common feature in cancer. Numerous studies indicate that persistent activation of the phosphoinositide 3-kinase (PI3K) pathway is often observed in cancer cells. 3-phosphoinositide dependent protein kinase-1 (PDK1), a transducer protein that functions downstream of PI3K, is responsible for the regulation of cell proliferation and migration and it also has been found to play a key role in different cancers, pancreatic and breast cancer amongst others. As PI3K is being described to be aberrantly expressed in several cancer types, designing inhibitors targeting various downstream molecules of PI3K has been the focus of anticancer agent development for a long time.
    [Show full text]
  • Inhibition of Aurora B Kinase Sensitizes a Subset of Human Glioma Cells to TRAIL Concomitant with Induction of TRAIL-R2
    Cell Death and Differentiation (2009) 16, 498–511 & 2009 Macmillan Publishers Limited All rights reserved 1350-9047/09 $32.00 www.nature.com/cdd Inhibition of Aurora B kinase sensitizes a subset of human glioma cells to TRAIL concomitant with induction of TRAIL-R2 JLi1, MG Anderson2, LA Tucker1, Y Shen1, KB Glaser1 and OJ Shah*,1 Small-molecule inhibitors of the Aurora A and B kinases interfere with mitotic centrosome function and disrupt the mitotic spindle assembly checkpoint resulting in polyploidization and apoptosis of proliferating cells. As such, several Aurora kinase inhibitors are at various stages of clinical development as anticancer agents. To identify candidate apoptosis-sensitizing genes that could be exploited in combination with Aurora kinase inhibitors in malignant glioma, we have carried out global gene expression analysis in a D54MG glioma cell derivative treated with three Aurora kinase inhibitors chosen for their distinctive selectivities: MLN8054 (Aurora A-selective), AZD1152 (Aurora B-selective), and VX-680 (Aurora A/B). The modulation of apoptotic gene expression by p53 under these conditions was ascertained, as p53 expression can be toggled on and off in this D54MG derivative by virtue of a stable, inducible, p53-targeting short hairpin RNA (D54MGshp53). This analysis identified the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) death receptor, TRAIL receptor 2 (TRAIL-R2), as an apoptosis- sensitizing gene induced selectively following inhibition of Aurora B. In glioma cell lines where TRAIL-R2 was induced following polyploidization, the sensitivity, kinetics, and magnitude of TRAIL-mediated apoptosis were enhanced. Our data shed light on the apoptotic program induced during polyploidization and suggest that TRAIL-R2 activation is a putative point of therapeutic intervention in combination with inhibitors of Aurora B.
    [Show full text]
  • A Framework for Identification of Actionable Cancer Genome
    A framework for identification of actionable cancer genome dependencies in small cell lung cancer Martin L. Sosa,b,c,d,1,2, Felix Dietleina,b,1, Martin Peifera,b, Jakob Schöttlea,b, Hyatt Balke-Wanta,b, Christian Müllera,b, Mirjam Kokera,b, André Richterse,f, Stefanie Heyncka,b, Florian Malchersa,b, Johannes M. Heuckmanna,b, Danila Seidela,b, Patrick A. Eyersg, Roland T. Ullrichb, Andrey P. Antonchickh, Viktor V. Vintonyakh, Peter M. Schneideri, Takashi Ninomiyaj, Herbert Waldmanne,h, Reinhard Büttnerk, Daniel Rauhe,f, Lukas C. Heukampk, and Roman K. Thomasa,b,k,2 aDepartment of Translational Genomics, University of Cologne, 50931 Cologne, Germany; bMax Planck Institute for Neurological Research, 50931 Cologne, Germany; cHoward Hughes Medical Institute, dDepartment of Cellular and Molecular Pharmacology, University of California, San Francisco, CA 94158; eChemical Genomics Center of the Max Planck Society, 44227 Dortmund, Germany; fTechnical University Dortmund, D-44221 Dortmund, Germany; gYorkshire Cancer Research (YCR) Institute for Cancer Studies, Cancer Research United Kingdom (CR-UK)/YCR Sheffield Cancer Research Centre, Department of Oncology, University of Sheffield, Sheffield S10 2RX, United Kingdom; hMax Planck Institute of Molecular Physiology, D-44227 Dortmund, Germany; iInstitute of Forensic Medicine, University of Cologne, 50823 Cologne, Germany; jDepartment of Hematology, Oncology, and Respiratory Medicine, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, 700-8558 Okayama, Japan; and kInstitute of Pathology, University of Cologne, 50924 Cologne, Germany Edited by Peter K. Vogt, The Scripps Research Institute, La Jolla, CA, and approved September 11, 2012 (received for review April 30, 2012) Small cell lung cancer (SCLC) accounts for about 15% of all lung and “synthetic lethality” have emerged (15–19).
    [Show full text]
  • (12) United States Patent (10) Patent No.: US 8,633,210 B2 Us M
    USOO863321 OB2 (12) United States Patent (10) Patent No.: US 8,633,210 B2 Charrier et al. (45) Date of Patent: *Jan. 21, 2014 (54) TRIAZOLE COMPOUNDS USEFUL AS 5,710, 158 A 1/1998 Myers et al. PROTEINKNASE INHIBITORS (Continued) (75) Inventors: Jean-Damien Charrier, Wantage (GB); FOREIGN PATENT DOCUMENTS Pan Li, Lexington, MA (US); Ronald Knegtel, Abingdon (GB); Julian DE 2458965 6, 1976 Marian Charles Golec, Faringdon E. 09: k l 3. (GB); David Bebbington, Newbury EP O3O2312 2, 1989 (GB); Hayley Marie Binch, Encinitas, GB 205.2487 1, 1981 CA (US) JP 10-130 150 5, 1998 JP 2000-026421 1, 2000 (73) Assignee: Vertex Pharmaceuticals Incorporated, wo E. gig. Cambridge, MA (US) WO 9322681 11, 1993 (*) Notice: Subject to any disclaimer, the term of this (Continued) patent is extended or adjusted under 35 U.S.C. 154(b) by 0 days. OTHER PUBLICATIONS This patent is Subject to a terminal dis- Wolff et, al., “Burger's Medicinal Chemistry and Drug Discovery.” claimer. 5th Ed. Part 1, pp.975-977 (1995).* Banker, et. al., (1996), Modern Pharmaceuticals, p. 596.* (21) Appl. No.: 13/094, 183 (1980),Ivashchenko, (12), 1673-7.*et al. Khimiya Geterotsiklicheskikh Soedinenii (22) Filed: Apr. 26, 2011 (Continued) (65) Prior Publication Data Primary Examiner — Jeffrey Murray US 2012/OO71657 A1 Mar. 22, 2012 (74) Attorney, Agent, or Firm — Rory C. Stewart Related U.S. Application Data (57) ABSTRACT (62) Division of application No. 1 1/492.450, filed on Jul. This invention describes novel triazole compounds of for 25, 2006, now Pat. No. 7,951,820, which is a division mula IX: of application No.
    [Show full text]