The Investigational Aurora Kinase A Inhibitor MLN8237 Induces Defects in Cell Viability and Cell-Cycle Progression in Malignant Bladder Cancer Cells In Vitro and In Vivo

Purpose: Despite more than 70,000 new cases of bladder cancer in the United States annually, patients with advanced disease have a poor prognosis due to limited treatment modalities. We evaluated Aurora kinase A, identified as an upregulated candidate molecule in bladder cancer, as a potential therapeutic target. Experimental Design: Gene expression in human bladder cancer samples was evaluated using RNA microarray and quantitative reverse transcriptase PCR. Effects of the Aurora kinase A inhibitor MLN8237 (Millennium) on cell dynamics in malignant T24 and UM-UC-3 and papilloma-derived RT4 bladder cells were evaluated in vitro and in vivo in a mouse xenograft model. Results: A set of 13 genes involved in the mitotic spindle checkpoint, including Aurora kinases A and B, were upregulated in human urothelial carcinoma compared with normal urothelium. The Aurora kinase A inhibitor MLN8237 induced cell-cycle arrest, aneuploidy, mitotic spindle failure, and apoptosis in the human bladder cancer cell lines T24 and UM-UC-3. MLN8237 also arrested tumor growth when administered orally over 4 weeks in a mouse bladder cancer xenograft model. Finally, in vitro sequential administration of MLN8237 with either paclitaxel or gemcitabine resulted in synergistic cytotoxic effects in T24 cells. Conclusions: Mitotic spindle checkpoint dysfunction is a common characteristic of human urothelial carcinoma and can be exploited with pharmacologic Aurora A inhibition. Given our demonstration of the ability of the Aurora A inhibitor MLN8237 to inhibit growth of bladder cancer in vitro and in vivo, we conclude that Aurora kinase inhibitors warrant further therapeutic investigation in bladder cancer. Clin Cancer Res; 19(7); 1717–28. ©2013 AACR.

[1]  H. Katayama,et al.  Mitotic kinase expression and colorectal cancer progression. , 1999, Journal of the National Cancer Institute.

[2]  R. Dreicer,et al.  Systemic chemotherapy for advanced bladder cancer: update and controversies. , 2006, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[3]  R. Vibhakar,et al.  Targeting Aurora Kinase A enhances radiation sensitivity of atypical teratoid rhabdoid tumor cells , 2012, Journal of Neuro-Oncology.

[4]  Jeffrey Ecsedy,et al.  The Aurora Kinase A Inhibitor MLN8237 Enhances Cisplatin-Induced Cell Death in Esophageal Adenocarcinoma Cells , 2012, Molecular Cancer Therapeutics.

[5]  H. Katayama,et al.  Increased mitotic phosphorylation of histone H3 attributable to AIM-1/Aurora-B overexpression contributes to chromosome number instability. , 2002, Cancer research.

[6]  L. Rimsza,et al.  Aurora inhibitor MLN8237 in combination with docetaxel enhances apoptosis and anti-tumor activity in mantle cell lymphoma. , 2011, Biochemical pharmacology.

[7]  Shuji Ogino,et al.  Aurora-A expression is independently associated with chromosomal instability in colorectal cancer. , 2009, Neoplasia.

[8]  W. Earnshaw,et al.  The cellular geography of Aurora kinases , 2003, Nature Reviews Molecular Cell Biology.

[9]  Benjamin M. Bolstad,et al.  affy - analysis of Affymetrix GeneChip data at the probe level , 2004, Bioinform..

[10]  S. Schwartz,et al.  p53 mutations in bladder carcinoma cell lines. , 1994, Oncology research.

[11]  P. Albers,et al.  Gemcitabine and cisplatin versus methotrexate, vinblastine, doxorubicin, and cisplatin in advanced or metastatic bladder cancer: results of a large, randomized, multinational, multicenter, phase III study. , 2000, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[12]  Alex E. Lash,et al.  Gene Expression Omnibus: NCBI gene expression and hybridization array data repository , 2002, Nucleic Acids Res..

[13]  L. Collette,et al.  Seven year update of an EORTC phase III trial of high-dose intensity M-VAC chemotherapy and G-CSF versus classic M-VAC in advanced urothelial tract tumours. , 2006, European journal of cancer.

[14]  R. Myers,et al.  Evolving gene/transcript definitions significantly alter the interpretation of GeneChip data , 2005, Nucleic acids research.

[15]  Takuya Kumazawa,et al.  Critical role of the nucleolus in activation of the p53-dependent postmitotic checkpoint. , 2011, Biochemical and biophysical research communications.

[16]  Shipra Agrawal,et al.  Biology of Aurora A kinase: Implications in cancer manifestation and therapy , 2011, Medicinal research reviews.

[17]  Luigi Cormio,et al.  Expression of mitotic kinases phospho-aurora A and aurora B correlates with clinical and pathological parameters in bladder neoplasms. , 2010, Histology and histopathology.

[18]  J. Berlin,et al.  Aurora Kinase Inhibitors - Rising Stars in Cancer Therapeutics? , 2010, Molecular Cancer Therapeutics.

[19]  A. Dar,et al.  Aurora kinase A inhibition leads to p73-dependent apoptosis in p53-deficient cancer cells. , 2008, Cancer research.

[20]  Terence P. Speed,et al.  A comparison of normalization methods for high density oligonucleotide array data based on variance and bias , 2003, Bioinform..

[21]  Olivier Cussenot,et al.  Gene expression study of Aurora-A reveals implication during bladder carcinogenesis and increasing values in invasive urothelial cancer. , 2008, Urology.

[22]  C. Ching,et al.  Mammalian target of rapamycin (mTOR) regulates cellular proliferation and tumor growth in urothelial carcinoma. , 2010, The American journal of pathology.

[23]  E. Sakamoto-Hojo,et al.  Cell cycle arrest and apoptosis in TP53 subtypes of bladder carcinoma cell lines treated with cisplatin and gemcitabine , 2010, Experimental biology and medicine.

[24]  Jeffrey Ecsedy,et al.  A novel Aurora-A kinase inhibitor MLN8237 induces cytotoxicity and cell-cycle arrest in multiple myeloma. , 2009, Blood.

[25]  A. Jemal,et al.  Cancer statistics, 2012 , 2012, CA: a cancer journal for clinicians.

[26]  D. Medina,et al.  Centrosome amplification and overexpression of aurora A are early events in rat mammary carcinogenesis. , 2002, Cancer research.

[27]  F. Giles,et al.  The novel Aurora A kinase inhibitor MLN8237 is active in resistant chronic myeloid leukaemia and significantly increases the efficacy of nilotinib , 2011, Journal of cellular and molecular medicine.

[28]  L. Margaritis,et al.  Thymidylate synthase inhibition induces p53-dependent and p53-independent apoptotic responses in human urinary bladder cancer cells , 2011, Journal of Cancer Research and Clinical Oncology.

[29]  Rafael A Irizarry,et al.  Exploration, normalization, and summaries of high density oligonucleotide array probe level data. , 2003, Biostatistics.

[30]  H. Katayama,et al.  Multinuclearity and increased ploidy caused by overexpression of the aurora- and Ipl1-like midbody-associated protein mitotic kinase in human cancer cells. , 1998, Cancer research.

[31]  A. Venkitaraman,et al.  AURORA-A amplification overrides the mitotic spindle assembly checkpoint, inducing resistance to Taxol. , 2003, Cancer cell.

[32]  F. Giles,et al.  Targeting Aurora A kinase activity with the investigational agent alisertib increases the efficacy of cytarabine through a FOXO‐dependent mechanism , 2012, International journal of cancer.

[33]  S. Shieh,et al.  TTK/hMps1 Mediates the p53-Dependent Postmitotic Checkpoint by Phosphorylating p53 at Thr18 , 2009, Molecular and Cellular Biology.

[34]  M. Mancini,et al.  Aurora-C kinase supports mitotic progression in the absence of Aurora-B , 2009, Cell cycle.

[35]  Geert J. P. L. Kops,et al.  On the road to cancer: aneuploidy and the mitotic checkpoint , 2005, Nature Reviews Cancer.

[36]  D. Pectasides,et al.  Current chemotherapeutic options for the treatment of advanced bladder cancer: a review. , 2013, Urologic oncology.

[37]  C. Cordon-Cardo,et al.  Inactivation of p53 and Pten promotes invasive bladder cancer. , 2009, Genes & development.

[38]  R. Fisher,et al.  Aurora A Inhibitor (MLN8237) plus Vincristine plus Rituximab Is Synthetic Lethal and a Potential Curative Therapy in Aggressive B-cell Non-Hodgkin Lymphoma , 2012, Clinical Cancer Research.

[39]  N. Dubrawsky.  Cancer statistics , 1989, CA: a cancer journal for clinicians.

[40]  R. Fucini,et al.  The Aurora kinase inhibitor SNS-314 shows broad therapeutic potential with chemotherapeutics and synergy with microtubule-targeted agents in a colon carcinoma model , 2009, Molecular Cancer Therapeutics.

[41]  H. Saya,et al.  Aurora-A — A guardian of poles , 2005, Nature Reviews Cancer.

[42]  Linjie Guo,et al.  The ATM–p53 pathway suppresses aneuploidy-induced tumorigenesis , 2010, Proceedings of the National Academy of Sciences.

[43]  N. Mori,et al.  Aurora A selective inhibitor MLN8237 suppresses the growth and survival of HTLV‐1‐infected T‐cells in vitro , 2010, Cancer science.

[44]  Sun Yan,et al.  Prognostic significance of Aurora-A expression in human bladder cancer. , 2011, Acta histochemica.

[45]  R. Cheney,et al.  Raf1, Aurora-A/STK15 and E-cadherin biomarkers expression in patients with pTa/pT1 urothelial bladder carcinoma; a retrospective TMA study of 246 patients with long-term follow-up. , 2006, European journal of surgical oncology : the journal of the European Society of Surgical Oncology and the British Association of Surgical Oncology.

[46]  Jian Kuang,et al.  Tumour amplified kinase STK15/BTAK induces centrosome amplification, aneuploidy and transformation , 1998, Nature Genetics.

[47]  C. Deng,et al.  Overexpression of aurora kinase A in mouse mammary epithelium induces genetic instability preceding mammary tumor formation , 2006, Oncogene.

[48]  T. Speed,et al.  Summaries of Affymetrix GeneChip probe level data. , 2003, Nucleic acids research.

[49]  Kaori Sasai,et al.  Aurora-C kinase is a novel chromosomal passenger protein that can complement Aurora-B kinase function in mitotic cells. , 2004, Cell motility and the cytoskeleton.

[50]  T. Chou.  Drug combination studies and their synergy quantification using the Chou-Talalay method. , 2010, Cancer research.

[51]  V. Paradis,et al.  Aurora-A/STK-15 is a predictive factor for recurrent behaviour in non-invasive bladder carcinoma: a study of 128 cases of non-invasive neoplasms , 2007, Virchows Archiv.