WEE1 Kinase Targeting Combined with DNA-Damaging Cancer Therapy Catalyzes Mitotic Catastrophe

WEE1 kinase is a key molecule in maintaining G2–cell-cycle checkpoint arrest for premitotic DNA repair. Whereas normal cells repair damaged DNA during G1-arrest, cancer cells often have a deficient G1-arrest and largely depend on G2-arrest. The molecular switch for the G2–M transition is held by WEE1 and is pushed forward by CDC25. WEE1 is overexpressed in various cancer types, including glioblastoma and breast cancer. Preclinical studies with cancer cell lines and animal models showed decreased cancer cell viability, reduced tumor burden, and improved survival after WEE1 inhibition by siRNA or small molecule inhibitors, which is enhanced by combination with conventional DNA-damaging therapy, such as radiotherapy and/or cytostatics. Mitotic catastrophe results from premature entry into mitosis with unrepaired lethal DNA damage. As such, cancer cells become sensitized to conventional therapy by WEE1 inhibition, in particular those with insufficient G1-arrest due to deficient p53 signaling, like glioblastoma cells. One WEE1 inhibitor has now reached clinical phase I studies. Dose-limiting toxicity consisted of hematologic events, nausea and/or vomiting, and fatigue. The combination of DNA-damaging cancer therapy with WEE1 inhibition seems to be a rational approach to push cancer cells in mitotic catastrophe. Its safety and efficacy are being evaluated in clinical studies. Clin Cancer Res; 17(13); 4200–7. ©2011 AACR.

[1]  P. Russell,et al.  Cell cycle regulation of human WEE1. , 1995, The EMBO journal.

[2]  Tsuyoshi Arai,et al.  Small-molecule inhibition of Wee1 kinase by MK-1775 selectively sensitizes p53-deficient tumor cells to DNA-damaging agents , 2009, Molecular Cancer Therapeutics.

[3]  J. Janetka,et al.  Keeping checkpoint kinases in line: new selective inhibitors in clinical trials. , 2008, Expert opinion on investigational drugs.

[4]  Daniel F Ortwine,et al.  4-Phenylpyrrolo[3,4-c]carbazole-1,3(2H,6H)-dione inhibitors of the checkpoint kinase Wee1. Structure-activity relationships for chromophore modification and phenyl ring substitution. , 2006, Journal of medicinal chemistry.

[5]  M. Gonen,et al.  Phase I trial of the cyclin-dependent kinase inhibitor and protein kinase C inhibitor 7-hydroxystaurosporine in combination with Fluorouracil in patients with advanced solid tumors. , 2005, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[6]  C. Hudis,et al.  Combination of trastuzumab and tanespimycin (17-AAG, KOS-953) is safe and active in trastuzumab-refractory HER-2 overexpressing breast cancer: a phase I dose-escalation study. , 2007, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[7]  H. Koga,et al.  Cell cycle regulation by the Wee1 Inhibitor PD0166285, Pyrido [2,3-d] pyimidine, in the B16 mouse melanoma cell line , 2006, BMC Cancer.

[8]  R. Mirimanoff,et al.  MGMT gene silencing and benefit from temozolomide in glioblastoma. , 2005, The New England journal of medicine.

[9]  Daniela S Krause,et al.  Tyrosine kinases as targets for cancer therapy. , 2005, The New England journal of medicine.

[10]  E. Sausville,et al.  Phase I trial of 72-hour continuous infusion UCN-01 in patients with refractory neoplasms. , 2001, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[11]  S. Yamaguchi,et al.  Control Mechanism of the Circadian Clock for Timing of Cell Division in Vivo , 2003, Science.

[12]  M. Yanagida,et al.  HIV-1 Vpr induces cell cycle G2 arrest in fission yeast (Schizosaccharomyces pombe) through a pathway involving regulatory and catalytic subunits of PP2A and acting on both Wee1 and Cdc25. , 2001, Virology.

[13]  Z. Szallasi,et al.  A signature of chromosomal instability inferred from gene expression profiles predicts clinical outcome in multiple human cancers , 2006, Nature Genetics.

[14]  S. Davidson,et al.  A phase I and pharmacokinetic study of short infusions of UCN-01 in patients with refractory solid tumors. , 2005, Clinical cancer research : an official journal of the American Association for Cancer Research.

[15]  Paul Russell,et al.  Negative regulation of mitosis by wee1 +, a gene encoding a protein kinase homolog , 1987, Cell.

[16]  H. Hirai,et al.  MK-1775, a small molecule Wee1 inhibitor, enhances anti-tumor efficacy of various DNA-damaging agents, including 5-fluorouracil , 2010, Cancer biology & therapy.

[17]  P. Nurse,et al.  Chk1 is a wee1 kinase in the G2 DNA damage checkpoint inhibiting cdc2 by Y15 phosphorylation , 1997, The EMBO journal.

[18]  T. Lawrence,et al.  Wild-Type TP53 Inhibits G2-Phase Checkpoint Abrogation and Radiosensitization Induced by PD0166285, a WEE1 Kinase Inhibitor , 2002, Radiation research.

[19]  E. Small,et al.  Time to disease progression to evaluate a novel protein kinase C inhibitor, UCN‐01, in renal cell carcinoma , 2004, Cancer.

[20]  Tak-Hong Cheung,et al.  Transcriptional repression of WEE1 by Kruppel-like factor 2 is involved in DNA damage-induced apoptosis , 2005, Oncogene.

[21]  M. Barbacid,et al.  Cell cycle, CDKs and cancer: a changing paradigm , 2009, Nature Reviews Cancer.

[22]  Y. Wang,et al.  Binding of 14-3-3beta to the carboxyl terminus of Wee1 increases Wee1 stability, kinase activity, and G2-M cell population. , 2000, Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research.

[23]  A. Kumagai,et al.  Positive regulation of Wee1 by Chk1 and 14-3-3 proteins. , 2001, Molecular biology of the cell.

[24]  A. Doherty,et al.  In vitro pharmacological characterization of PD 166285, a new nanomolar potent and broadly active protein tyrosine kinase inhibitor. , 1997, The Journal of pharmacology and experimental therapeutics.

[25]  W. Denny,et al.  Synthesis and structure-activity relationships of N-6 substituted analogues of 9-hydroxy-4-phenylpyrrolo[3,4-c]carbazole-1,3(2H,6H)-diones as inhibitors of Wee1 and Chk1 checkpoint kinases. , 2008, European journal of medicinal chemistry.

[26]  S. Kornbluth,et al.  Cdc25 and Wee1: analogous opposites? , 2007, Cell Division.

[27]  T. Chou,et al.  90-kDa Heat Shock Protein Inhibition Abrogates the Topoisomerase I Poison-Induced G2/M Checkpoint in p53-Null Tumor Cells by Depleting Chk1 and Wee1 , 2009, Molecular Pharmacology.

[28]  P. C. de Witt Hamer Small molecule kinase inhibitors in glioblastoma: a systematic review of clinical studies. , 2010, Neuro-oncology.

[29]  J. Sebolt-Leopold,et al.  Knockdown of Chk1, Wee1 and Myt1 by RNA Interference Abrogates G2 Checkpoint and Induces Apoptosis , 2004, Cancer biology & therapy.

[30]  E. Baker,et al.  Structure and inhibition of the human cell cycle checkpoint kinase, Wee1A kinase: an atypical tyrosine kinase with a key role in CDK1 regulation. , 2005, Structure.

[31]  Guido Kroemer,et al.  Cell death by mitotic catastrophe: a molecular definition , 2004, Oncogene.

[32]  J. Levine,et al.  Surfing the p53 network , 2000, Nature.

[33]  R. Medema,et al.  Human prostate epithelium lacks Wee1A-mediated DNA damage-induced checkpoint enforcement , 2007, Proceedings of the National Academy of Sciences.

[34]  P. Richardson,et al.  Tanespimycin monotherapy in relapsed multiple myeloma: results of a phase 1 dose‐escalation study , 2010, British journal of haematology.

[35]  Thomas Dandekar,et al.  Explorative data analysis of MCL reveals gene expression networks implicated in survival and prognosis supported by explorative CGH analysis , 2008, BMC Cancer.

[36]  W. Denny,et al.  Structure-activity relationships for 2-anilino-6-phenylpyrido[2,3-d]pyrimidin-7(8H)-ones as inhibitors of the cellular checkpoint kinase Wee1. , 2005, Bioorganic & medicinal chemistry letters.

[37]  S. Ramalingam,et al.  A Phase I Study of 17-Allylamino-17-Demethoxygeldanamycin Combined with Paclitaxel in Patients with Advanced Solid Malignancies , 2008, Clinical Cancer Research.

[38]  Valerie Cavett,et al.  Activation Domain-dependent Degradation of Somatic Wee1 Kinase* , 2009, The Journal of Biological Chemistry.

[39]  L. Neckers,et al.  Hsp90 phosphorylation, Wee1 and the cell cycle , 2010, Cell cycle.

[40]  H. Kuwano,et al.  The clinical significance of Cyclin B1 and Wee1 expression in non-small-cell lung cancer. , 2004, Annals of oncology : official journal of the European Society for Medical Oncology.

[41]  J. Aten,et al.  In silico analysis of kinase expression identifies WEE1 as a gatekeeper against mitotic catastrophe in glioblastoma. , 2010, Cancer cell.

[42]  F. McKeon,et al.  Human wee1 maintains mitotic timing by protecting the nucleus from cytoplasmically activated cdc2 kinase , 1993, Cell.

[43]  Rong Li,et al.  Mitotic progression becomes irreversible in prometaphase and collapses when Wee1 and Cdc25 are inhibited , 2011, Molecular biology of the cell.

[44]  Pierre Dubus,et al.  Cdk1 is sufficient to drive the mammalian cell cycle , 2007, Nature.

[45]  T. Dougherty,et al.  Anti-Angiogenic Activity of Selected Receptor Tyrosine Kinase Inhibitors, PD166285 and PD173074: Implications for Combination Treatment with Photodynamic Therapy , 2004, Investigational New Drugs.

[46]  U. Mansmann,et al.  Differential gene expression in colon carcinoma cells and tissues detected with a cDNA array , 1999, International journal of cancer.

[47]  A. Maitra,et al.  MK-1775, a Potent Wee1 Inhibitor, Synergizes with Gemcitabine to Achieve Tumor Regressions, Selectively in p53-Deficient Pancreatic Cancer Xenografts , 2011, Clinical Cancer Research.

[48]  H. Hirai,et al.  Discovery of gene expression-based pharmacodynamic biomarker for a p53 context-specific anti-tumor drug Wee1 inhibitor , 2009, Molecular Cancer.

[49]  Jun Yu Li,et al.  Radiosensitization of p53 mutant cells by PD0166285, a novel G(2) checkpoint abrogator. , 2001, Cancer research.

[50]  Yoshiaki Miyauchi,et al.  Cyclins and cyclin‐dependent kinases: Comparative study of hepatocellular carcinoma versus cirrhosis , 2003, Hepatology.

[51]  T. Tsuruo,et al.  Akt/Protein Kinase B-Dependent Phosphorylation and Inactivation of WEE1Hu Promote Cell Cycle Progression at G2/M Transition , 2005, Molecular and Cellular Biology.

[52]  Jiri Bartek,et al.  Targeting the checkpoint kinases: chemosensitization versus chemoprotection , 2004, Nature Reviews Cancer.

[53]  E. Sausville,et al.  Safety, Efficacy, Pharmacokinetics, and Pharmacodynamics of the Combination of Sorafenib and Tanespimycin , 2010, Clinical Cancer Research.

[54]  Elizabeth Iorns,et al.  Integrated Functional, Gene Expression and Genomic Analysis for the Identification of Cancer Targets , 2009, PloS one.

[55]  C. Britten,et al.  G2 checkpoint abrogation and checkpoint kinase-1 targeting in the treatment of cancer , 2008, British Journal of Cancer.

[56]  Paul Nurse,et al.  Genetic control of cell size at cell division in yeast , 1975, Nature.

[57]  L. Schwartz,et al.  A phase II trial of 17-(Allylamino)-17-demethoxygeldanamycin in patients with papillary and clear cell renal cell carcinoma , 2006, Investigational New Drugs.

[58]  H. Koga,et al.  Inhibition of proteasome‐dependent degradation of Wee1 in G2‐arrested Hep3B cells by TGFβ1 , 2003, Molecular carcinogenesis.

[59]  Joshua M. Korn,et al.  Comprehensive genomic characterization defines human glioblastoma genes and core pathways , 2008, Nature.

[60]  J. Wolchok,et al.  Phase II Trial of 17-Allylamino-17-Demethoxygeldanamycin in Patients with Metastatic Melanoma , 2008, Clinical Cancer Research.

[61]  M. Igarashi,et al.  Wee1 +-like gene in human cells , 1991, Nature.

[62]  A. Oza,et al.  A phase I pharmacological and pharmacodynamic study of MK-1775, a Wee1 tyrosine kinase inhibitor, in monotherapy and combination with gemcitabine, cisplatin, or carboplatin in patients with advanced solid tumors. , 2010 .

[63]  N Watanabe,et al.  Regulation of the human WEE1Hu CDK tyrosine 15‐kinase during the cell cycle. , 1995, The EMBO journal.

[64]  R. Medema,et al.  Polo-like kinase-1 controls recovery from a G2 DNA damage-induced arrest in mammalian cells. , 2004, Molecular cell.

[65]  C. Deng,et al.  Murine Wee1 Plays a Critical Role in Cell Cycle Regulation and Pre-Implantation Stages of Embryonic Development , 2006, International journal of biological sciences.

[66]  H. Dixon,et al.  Therapeutic Exploitation of Checkpoint Defects in Cancer Cells Lacking p53 Function , 2002, Cell cycle.

[67]  F. Sarkar,et al.  A phase II trial of 17-allylamino-17- demethoxygeldanamycin in patients with hormone-refractory metastatic prostate cancer. , 2005, Clinical prostate cancer.

[68]  M. Fallahi,et al.  Redundant Ubiquitin Ligase Activities Regulate Wee1 Degradation and Mitotic Entry , 2007, Cell cycle.

[69]  H. Yoshida,et al.  Akt inhibits Myt1 in the signalling pathway that leads to meiotic G2/M-phase transition , 2002, Nature Cell Biology.

[70]  H. Piwnica-Worms,et al.  Inactivation of the p34cdc2-cyclin B complex by the human WEE1 tyrosine kinase. , 1992, Science.

[71]  C. Erlichman Tanespimycin: the opportunities and challenges of targeting heat shock protein 90 , 2009, Expert opinion on investigational drugs.

[72]  N. Caplen,et al.  Identification of WEE1 as a potential molecular target in cancer cells by RNAi screening of the human tyrosine kinome , 2010, Breast Cancer Research and Treatment.

[73]  Yi Sun,et al.  Binding of 14-3-3 b to the Carboxyl Terminus of Wee 1 Increases Wee 1 Stability , Kinase Activity , and G 2M Cell Population , 2000 .

[74]  M. Ernstoff,et al.  Modulation of Cell Cycle Progression in Human Tumors: A Pharmacokinetic and Tumor Molecular Pharmacodynamic Study of Cisplatin Plus the Chk1 Inhibitor UCN-01 (NSC 638850) , 2006, Clinical Cancer Research.

[75]  S. Taviaux,et al.  Localization of human cell cycle regulatory genes CDC25C to 5q31 and WEE1 to 11p15.3-11p15.1 by fluorescence in situ hybridization. , 1993, Genomics.

[76]  Mark W. Dewhirst,et al.  Glioma stem cells promote radioresistance by preferential activation of the DNA damage response , 2006, Nature.

[77]  Bernard Ducommun,et al.  CDC25 phosphatases in cancer cells: key players? Good targets? , 2007, Nature Reviews Cancer.

[78]  W. Hait,et al.  Reversal of Stathmin-Mediated Resistance to Paclitaxel and Vinblastine in Human Breast Carcinoma Cells , 2007, Molecular Pharmacology.

[79]  James E. Ferrell,et al.  Substrate Competition as a Source of Ultrasensitivity in the Inactivation of Wee1 , 2007, Cell.

[80]  Philip C. De Witt Hamer,et al.  Small molecule kinase inhibitors in glioblastoma: a systematic review of clinical studies , 2010 .

[81]  J. Ruderman,et al.  Changes in regulatory phosphorylation of Cdc25C Ser287 and Wee1 Ser549 during normal cell cycle progression and checkpoint arrests. , 2005, Molecular biology of the cell.

[82]  Hiroyuki Osada,et al.  M-phase kinases induce phospho-dependent ubiquitination of somatic Wee1 by SCFbeta-TrCP. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[83]  A. Levine,et al.  Surfing the p53 network , 2000, Nature.

[84]  W. Denny,et al.  Synthesis and structure-activity relationships of soluble 8-substituted 4-(2-chlorophenyl)-9-hydroxypyrrolo[3,4-c]carbazole-1,3(2H,6H)-diones as inhibitors of the Wee1 and Chk1 checkpoint kinases. , 2008, Bioorganic & medicinal chemistry letters.

[85]  T. Kawabe G2 checkpoint abrogators as anticancer drugs. , 2004, Molecular cancer therapeutics.