The tyrosine kinase inhibitor nintedanib activates SHP-1 and induces apoptosis in triple-negative breast cancer cells

Triple-negative breast cancer (TNBC) remains difficult to treat and urgently needs new therapeutic options. Nintedanib, a multikinase inhibitor, has exhibited efficacy in early clinical trials for HER2-negative breast cancer. In this study, we examined a new molecular mechanism of nintedanib in TNBC. The results demonstrated that nintedanib enhanced TNBC cell apoptosis, which was accompanied by a reduction of p-STAT3 and its downstream proteins. STAT3 overexpression suppressed nintedanib-mediated apoptosis and further increased the activity of purified SHP-1 protein. Moreover, treatment with either a specific inhibitor of SHP-1 or SHP-1-targeted siRNA reduced the apoptotic effects of nintedanib, which validates the role of SHP-1 in nintedanib-mediated apoptosis. Furthermore, nintedanib-induced apoptosis was attenuated in TNBC cells expressing SHP-1 mutants with constantly open conformations, suggesting that the autoinhibitory mechanism of SHP-1 attenuated the effects of nintedanib. Importantly, nintedanib significantly inhibited tumor growth via the SHP-1/p-STAT3 pathway. Clinically, SHP-1 levels were downregulated, whereas p-STAT3 was upregulated in tumor tissues, and SHP-1 transcripts were associated with improved disease-free survival in TNBC patients. Our findings revealed that nintedanib induces TNBC apoptosis by acting as a SHP-1 agonist, suggesting that targeting STAT3 by enhancing SHP-1 expression could be a viable therapeutic strategy against TNBC.

[1]  J. Rodríguez-Ubreva,et al.  SHP-1 in cell-cycle regulation. , 2011, Anti-cancer agents in medicinal chemistry.

[2]  Balazs Györffy,et al.  Meta-analysis of gene expression profiles related to relapse-free survival in 1,079 breast cancer patients , 2009, Breast Cancer Research and Treatment.

[3]  B. Goh,et al.  Targeting transcription factor STAT3 for cancer prevention and therapy. , 2016, Pharmacology & therapeutics.

[4]  Z. Zhao,et al.  Crystal Structure of Human Protein-tyrosine Phosphatase SHP-1* , 2003, The Journal of Biological Chemistry.

[5]  K. Siminovitch,et al.  Roles of the SHP-1 tyrosine phosphatase in the negative regulation of cell signalling. , 2000, Seminars in immunology.

[6]  R. Brekken,et al.  Nintedanib, a triple angiokinase inhibitor, enhances cytotoxic therapy response in pancreatic cancer. , 2015, Cancer letters.

[7]  A. Poole,et al.  A SHPing tale: perspectives on the regulation of SHP-1 and SHP-2 tyrosine phosphatases by the C-terminal tail. , 2005, Cellular signalling.

[8]  X. Chen,et al.  Identification of human triple-negative breast cancer subtypes and preclinical models for selection of targeted therapies. , 2011, The Journal of clinical investigation.

[9]  D. Frank,et al.  Distinct roles of STAT3 and STAT5 in the pathogenesis and targeted therapy of breast cancer , 2014, Molecular and Cellular Endocrinology.

[10]  Kuen-Feng Chen,et al.  SHP-1 is a target of regorafenib in colorectal cancer , 2014, Oncotarget.

[11]  H. Seo,et al.  Trichosanthes kirilowii Ethanol Extract and Cucurbitacin D Inhibit Cell Growth and Induce Apoptosis through Inhibition of STAT3 Activity in Breast Cancer Cells , 2013, Evidence-based complementary and alternative medicine : eCAM.

[12]  Kuen-Feng Chen,et al.  Novel sorafenib analogues induce apoptosis through SHP-1 dependent STAT3 inactivation in human breast cancer cells , 2013, Breast Cancer Research.

[13]  Z. Zhao,et al.  Crystal Structure of the Catalytic Domain of Protein-tyrosine Phosphatase SHP-1* , 1998, The Journal of Biological Chemistry.

[14]  N. Vaysse,et al.  sst2 Somatostatin Receptor Mediates Negative Regulation of Insulin Receptor Signaling through the Tyrosine Phosphatase SHP-1* , 1998, The Journal of Biological Chemistry.

[15]  Pei-Yi Chu,et al.  Disrupting VEGF-A paracrine and autocrine loops by targeting SHP-1 suppresses triple negative breast cancer metastasis , 2016, Scientific Reports.

[16]  B. Yadav,et al.  Biomarkers in triple negative breast cancer: A review. , 2015, World journal of clinical oncology.

[17]  Kuen-Feng Chen,et al.  RFX1–dependent activation of SHP-1 induces autophagy by a novel obatoclax derivative in hepatocellular carcinoma cells , 2014, Oncotarget.

[18]  Shishinn Sun,et al.  PTEN is a negative regulator of STAT3 activation in human papillomavirus-infected cells. , 2002, The Journal of general virology.

[19]  Yiling Lu,et al.  SHP-1 Regulates Lck-induced Phosphatidylinositol 3-Kinase Phosphorylation and Activity* , 1999, The Journal of Biological Chemistry.

[20]  A. Masamune,et al.  Activation of JAK-STAT pathway is required for platelet-derived growth factor-induced proliferation of pancreatic stellate cells. , 2005, World journal of gastroenterology.

[21]  D. Gerber,et al.  BIBF 1120 (Nintedanib), a Triple Angiokinase Inhibitor, Induces Hypoxia but not EMT and Blocks Progression of Preclinical Models of Lung and Pancreatic Cancer , 2013, Molecular Cancer Therapeutics.

[22]  M. Reck.  Nintedanib: examining the development and mechanism of action of a novel triple angiokinase inhibitor , 2015, Expert review of anticancer therapy.

[23]  M. Quinn,et al.  Targeted Disruption of the JAK2/STAT3 Pathway in Combination with Systemic Administration of Paclitaxel Inhibits the Priming of Ovarian Cancer Stem Cells Leading to a Reduced Tumor Burden , 2014, Front. Oncol..

[24]  Kuen-Feng Chen,et al.  Synthesis and biological activity of obatoclax derivatives as novel and potent SHP-1 agonists. , 2012, European journal of medicinal chemistry.

[25]  A. Urruticoechea,et al.  Phase I clinical trial of nintedanib plus paclitaxel in early HER-2-negative breast cancer (CNIO-BR-01-2010/GEICAM-2010-10 study) , 2014, British Journal of Cancer.

[26]  W. Sommergruber,et al.  BIBF 1120: triple angiokinase inhibitor with sustained receptor blockade and good antitumor efficacy. , 2008, Cancer research.

[27]  Kuen-Feng Chen,et al.  A novel obatoclax derivative, SC-2001, induces apoptosis in hepatocellular carcinoma cells through SHP-1-dependent STAT3 inactivation. , 2012, Cancer letters.

[28]  Alexander Pautsch,et al.  Mode of action of nintedanib in the treatment of idiopathic pulmonary fibrosis , 2015, European Respiratory Journal.

[29]  Debra L Winkeljohn.  Triple-negative breast cancer. , 2008, Clinical journal of oncology nursing.

[30]  Kuen-Feng Chen,et al.  Signal transducer and activator of transcription 3 is a major kinase-independent target of sorafenib in hepatocellular carcinoma. , 2011, Journal of hepatology.

[31]  L. Platanias,et al.  Direct association with and dephosphorylation of Jak2 kinase by the SH2-domain-containing protein tyrosine phosphatase SHP-1 , 1996, Molecular and cellular biology.

[32]  D. Zopf,et al.  Regorafenib (BAY 73‐4506): A new oral multikinase inhibitor of angiogenic, stromal and oncogenic receptor tyrosine kinases with potent preclinical antitumor activity , 2011, International journal of cancer.

[33]  Soo-Jin Jeong,et al.  Oral administration of penta-O-galloyl-β-D-glucose suppresses triple-negative breast cancer xenograft growth and metastasis in strong association with JAK1-STAT3 inhibition. , 2011, Carcinogenesis.

[34]  Jan Budczies,et al.  Online Survival Analysis Software to Assess the Prognostic Value of Biomarkers Using Transcriptomic Data in Non-Small-Cell Lung Cancer , 2013, PloS one.

[35]  Z. Szallasi,et al.  An online survival analysis tool to rapidly assess the effect of 22,277 genes on breast cancer prognosis using microarray data of 1,809 patients , 2010, Breast Cancer Research and Treatment.

[36]  R. Montironi,et al.  Investigational therapies targeting signal transducer and activator of transcription 3 for the treatment of cancer , 2015, Expert opinion on investigational drugs.

[37]  Hua Yu,et al.  Constitutive activation of Stat3 by the Src and JAK tyrosine kinases participates in growth regulation of human breast carcinoma cells , 2001, Oncogene.

[38]  B. Ryffel,et al.  Antifibrotic and Anti-inflammatory Activity of the Tyrosine Kinase Inhibitor Nintedanib in Experimental Models of Lung Fibrosis , 2014, The Journal of Pharmacology and Experimental Therapeutics.

[39]  D. Backos,et al.  Strategies and Approaches of Targeting STAT3 for Cancer Treatment. , 2016, ACS chemical biology.

[40]  H. Baumann,et al.  Protein Tyrosine Phosphatase 2 (SHP-2) Moderates Signaling by gp130 but Is Not Required for the Induction of Acute-Phase Plasma Protein Genes in Hepatic Cells , 1998, Molecular and Cellular Biology.

[41]  Preclinical S Tudy.  Meta-analysis of gene expression profiles related to relapse-free survival in 1,079 breast cancer patients , 2009 .

[42]  W. Stetler-Stevenson,et al.  TIMP1 induces CD44 expression and the activation and nuclear translocation of SHP1 during the late centrocyte/post-germinal center B cell differentiation. , 2008, Cancer letters.

[43]  C. Wu,et al.  SHP‐1 suppresses cancer cell growth by promoting degradation of JAK kinases , 2003, Journal of cellular biochemistry.

[44]  Kuen-Feng Chen,et al.  Nintedanib (BIBF-1120) inhibits hepatocellular carcinoma growth independent of angiokinase activity. , 2014, Journal of hepatology.

[45]  Vandana G. Abramson,et al.  New Strategies for Triple-Negative Breast Cancer—Deciphering the Heterogeneity , 2014, Clinical Cancer Research.

[46]  Kuen-Feng Chen,et al.  Dovitinib sensitizes hepatocellular carcinoma cells to TRAIL and tigatuzumab, a novel anti-DR5 antibody, through SHP-1-dependent inhibition of STAT3. , 2012, Biochemical pharmacology.

[47]  Kuen-Feng Chen,et al.  Dovitinib Induces Apoptosis and Overcomes Sorafenib Resistance in Hepatocellular Carcinoma through SHP-1–Mediated Inhibition of STAT3 , 2011, Molecular Cancer Therapeutics.

[48]  Kuen-Feng Chen,et al.  SC-2001 Overcomes STAT3-mediated Sorafenib Resistance through RFX-1/SHP-1 Activation in Hepatocellular Carcinoma , 2014, Neoplasia.

[49]  M. Marra,et al.  The PPAR-γ agonist troglitazone antagonizes survival pathways induced by STAT-3 in recombinant interferon-β treated pancreatic cancer cells. , 2012, Biotechnology advances.