Inhibition of the spindle assembly checkpoint kinase TTK enhances the efficacy of docetaxel in a triple-negative breast cancer model.

BACKGROUND Triple-negative breast cancers (TNBC) are considered the most aggressive type of breast cancer, for which no targeted therapy exists at the moment. These tumors are characterized by having a high degree of chromosome instability and often overexpress the spindle assembly checkpoint kinase TTK. To explore the potential of TTK inhibition as a targeted therapy in TNBC, we developed a highly potent and selective small molecule inhibitor of TTK, NTRC 0066-0. RESULTS AND CONCLUSIONS The compound is characterized by long residence time on the target and inhibits the proliferation of a wide variety of human cancer cell lines with potency in the same range as marketed cytotoxic agents. In cell lines and in mice, NTRC 0066-0 inhibits the phosphorylation of a TTK substrate and induces chromosome missegregation. NTRC 0066-0 inhibits tumor growth in MDA-MB-231 xenografts as a single agent after oral application. To address the effect of the inhibitor in breast cancer, we used a well-defined mouse model that spontaneously develops breast tumors that share key morphologic and molecular features with human TNBC. Our studies show that combination of NTRC 0066-0 with a therapeutic dose of docetaxel resulted in doubling of mouse survival and extended tumor remission, without toxicity. Furthermore, we observed that treatment efficacy is only achieved upon co-administration of the two compounds, which suggests a synergistic in vivo effect. Therefore, we propose TTK inhibition as a novel therapeutic target for neoadjuvant therapy in TNBC.

[1]  Chao Zhang,et al.  Mps1 directs the assembly of Cdc20 inhibitory complexes during interphase and mitosis to control M phase timing and spindle checkpoint signaling , 2010, The Journal of cell biology.

[2]  O. Kallioniemi,et al.  Analysis of Kinase Gene Expression Patterns across 5681 Human Tissue Samples Reveals Functional Genomic Taxonomy of the Kinome , 2010, PloS one.

[3]  R. Houlston,et al.  Association between chromosomal instability and prognosis in colorectal cancer: a meta-analysis , 2008, Gut.

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

[5]  R. Medema,et al.  Intravital FRET Imaging of Tumor Cell Viability and Mitosis during Chemotherapy , 2013, PloS one.

[6]  R. Medema,et al.  Elevating the frequency of chromosome mis-segregation as a strategy to kill tumor cells , 2009, Proceedings of the National Academy of Sciences.

[7]  Antonella Isacchi,et al.  Targeting the mitotic checkpoint for cancer therapy with NMS-P715, an inhibitor of MPS1 kinase. , 2010, Cancer research.

[8]  Emmanuel Barillot,et al.  TTK/hMPS1 Is an Attractive Therapeutic Target for Triple-Negative Breast Cancer , 2013, PloS one.

[9]  Xuedong Liu,et al.  The MPS1 family of protein kinases. , 2012, Annual review of biochemistry.

[10]  G. Mills,et al.  Expression of TTK, a novel human protein kinase, is associated with cell proliferation. , 1992, The Journal of biological chemistry.

[11]  Lajos Pusztai,et al.  Gene-expression signatures in breast cancer. , 2009, The New England journal of medicine.

[12]  E. Gabrielson,et al.  High levels of the Mps1 checkpoint protein are protective of aneuploidy in breast cancer cells , 2011, Proceedings of the National Academy of Sciences of the United States of America.

[13]  K. Yoshino,et al.  Comparison of the Cancer Gene Targeting and Biochemical Selectivities of All Targeted Kinase Inhibitors Approved for Clinical Use , 2014, PloS one.

[14]  J. Reis-Filho,et al.  Taxane benefit in breast cancer—a role for grade and chromosomal stability , 2013, Nature Reviews Clinical Oncology.

[15]  Mark E. Burkard,et al.  Cytotoxicity of Paclitaxel in Breast Cancer Is due to Chromosome Missegregation on Multipolar Spindles , 2014, Science Translational Medicine.

[16]  S. Wacholder,et al.  Gene Expression Signature of Cigarette Smoking and Its Role in Lung Adenocarcinoma Development and Survival , 2008, PloS one.

[17]  Junjie Chen,et al.  The mitotic checkpoint in cancer and aging: what have mice taught us? , 2005, Current opinion in cell biology.

[18]  L. Galluzzi,et al.  Characterization of novel MPS1 inhibitors with preclinical anticancer activity , 2013, Cell Death and Differentiation.

[19]  Andrea Musacchio,et al.  Dissecting the role of MPS1 in chromosome biorientation and the spindle checkpoint through the small molecule inhibitor reversine , 2010, The Journal of cell biology.

[20]  Mindy I. Davis,et al.  Comprehensive analysis of kinase inhibitor selectivity , 2011, Nature Biotechnology.

[21]  J Verweij,et al.  Clinical characteristics of severe peripheral neuropathy induced by docetaxel (Taxotere). , 1997, Annals of oncology : official journal of the European Society for Medical Oncology.

[22]  Shridar Ganesan,et al.  Loss of 53BP1 causes PARP inhibitor resistance in Brca1-mutated mouse mammary tumors. , 2013, Cancer discovery.

[23]  Taebo Sim,et al.  Small Molecule Kinase Inhibitors Provide Insight into Mps1 Cell Cycle Function , 2010, Nature chemical biology.

[24]  K. Kashfi,et al.  Abstract 4608: Hydrogen sulfide-releasing aspirin inhibits the growth of leukemic Jurkat cells and modulates β-catenin expression , 2011 .

[25]  K. Kinzler,et al.  Genetic instability in colorectal cancers , 1997, Nature.

[26]  M. Koltzenburg,et al.  Peripheral nerve damage associated with administration of taxanes in patients with cancer. , 2008, Critical reviews in oncology/hematology.

[27]  S. Lakhani,et al.  Meta-analysis of the global gene expression profile of triple-negative breast cancer identifies genes for the prognostication and treatment of aggressive breast cancer , 2014, Oncogenesis.

[28]  G. Kops,et al.  Negative feedback at kinetochores underlies a responsive spindle checkpoint signal , 2014, Nature Cell Biology.

[29]  Chao Zhang,et al.  Chemical Genetic Inhibition of Mps1 in Stable Human Cell Lines Reveals Novel Aspects of Mps1 Function in Mitosis , 2010, PloS one.

[30]  Jorge S Reis-Filho,et al.  Triple-negative breast cancer. , 2010, The New England journal of medicine.

[31]  E. Dı́az-Rodrı́guez,et al.  Complete loss of the tumor suppressor MAD2 causes premature cyclin B degradation and mitotic failure in human somatic cells. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[32]  Andrea Musacchio,et al.  Sustained Mps1 activity is required in mitosis to recruit O-Mad2 to the Mad1–C-Mad2 core complex , 2010, The Journal of cell biology.

[33]  Geert J P L Kops,et al.  Lethality to human cancer cells through massive chromosome loss by inhibition of the mitotic checkpoint. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[34]  M. Kris,et al.  Pulsatile Administration of the Epidermal Growth Factor Receptor Inhibitor Gefitinib Is Significantly More Effective than Continuous Dosing for Sensitizing Tumors to Paclitaxel , 2005, Clinical Cancer Research.

[35]  Eytan Domany,et al.  Association of survival and disease progression with chromosomal instability: A genomic exploration of colorectal cancer , 2009, Proceedings of the National Academy of Sciences.

[36]  Ralph Weissleder,et al.  Analysis of mitosis and antimitotic drug responses in tumors by in vivo microscopy and single-cell pharmacodynamics. , 2011, Cancer research.

[37]  Geert J P L Kops,et al.  Joined at the hip: kinetochores, microtubules, and spindle assembly checkpoint signaling. , 2015, Trends in cell biology.

[38]  J. Peterse,et al.  Somatic loss of BRCA1 and p53 in mice induces mammary tumors with features of human BRCA1-mutated basal-like breast cancer , 2007, Proceedings of the National Academy of Sciences.

[39]  Takashi Takahashi,et al.  Association between mitotic spindle checkpoint impairment and susceptibility to the induction of apoptosis by anti-microtubule agents in human lung cancers. , 2003, The American journal of pathology.

[40]  Yuya Yamagishi,et al.  MPS1/Mph1 phosphorylates the kinetochore protein KNL1/Spc7 to recruit SAC components , 2012, Nature Cell Biology.

[41]  Aaron Rogers,et al.  Characterization of the Cellular and Antitumor Effects of MPI-0479605, a Small-Molecule Inhibitor of the Mitotic Kinase Mps1 , 2011, Molecular Cancer Therapeutics.

[42]  Narendra Kumar Patel,et al.  Discovery of inhibitors of the mitotic kinase TTK based on N-(3-(3-sulfamoylphenyl)-1H-indazol-5-yl)-acetamides and carboxamides. , 2014, Bioorganic & medicinal chemistry.

[43]  M. Bittner,et al.  A cell proliferation and chromosomal instability signature in anaplastic thyroid carcinoma. , 2007, Cancer research.

[44]  Jos Jonkers,et al.  Selective induction of chemotherapy resistance of mammary tumors in a conditional mouse model for hereditary breast cancer , 2007, Proceedings of the National Academy of Sciences.

[45]  M. Korc,et al.  Selective Inhibition of Pancreatic Ductal Adenocarcinoma Cell Growth by the Mitotic MPS1 Kinase Inhibitor NMS-P715 , 2013, Molecular Cancer Therapeutics.

[46]  T. Lagerweij,et al.  Effects of the selective MPS1 inhibitor MPS1-IN-3 on glioblastoma sensitivity to antimitotic drugs. , 2013, Journal of the National Cancer Institute.

[47]  Ju-Hyung Woo,et al.  Increased Expression of Mitotic Checkpoint Genes in Breast Cancer Cells with Chromosomal Instability , 2006, Clinical Cancer Research.

[48]  W. Bornmann,et al.  Reversal of anticancer multidrug resistance by the ardeemins. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[49]  Dan-dan Wang,et al.  Expression and Function Analysis of Mitotic Checkpoint Genes Identifies TTK as a Potential Therapeutic Target for Human Hepatocellular Carcinoma , 2014, PloS one.