The efficacy of a novel, dual PI3K/mTOR inhibitor NVP‐BEZ235 to enhance chemotherapy and antiangiogenic response in pancreatic cancer

Gemcitabine has limited clinical benefits for pancreatic ductal adenocarcinoma (PDAC). The phosphatidylinositol‐3‐kinase (PI3K)/AKT and mammalian target of rapamycin (mTOR) signaling pathways are frequently dysregulated in PDAC. We investigated the effects of NVP‐BEZ235, a novel dual PI3K/mTOR inhibitor, in combination with gemcitabine and endothelial monocyte activating polypeptide II (EMAP) in experimental PDAC. Cell proliferation and protein expression were analyzed by WST‐1 assay and Western blotting. Animal survival experiments were performed in murine xenografts. BEZ235 caused a decrease in phospho‐AKT and phospho‐mTOR expression in PDAC (AsPC‐1), endothelial (HUVECs), and fibroblast (WI‐38) cells. BEZ235 inhibited in vitro proliferation of all four PDAC cell lines tested. Additive effects on proliferation inhibition were observed in the BEZ235‐gemcitabine combination in PDAC cells and in combination of BEZ235 or EMAP with gemcitabine in HUVECs and WI‐38 cells. BEZ235, alone or in combination with gemcitabine and EMAP, induced apoptosis in AsPC‐1, HUVECs, and WI‐38 cells as observed by increased expression of cleaved poly (ADP‐ribose) polymerase‐1 (PARP‐1) and caspase‐3 proteins. Compared to controls (median survival: 16 days), animal survival increased after BEZ235 and EMAP therapy alone (both 21 days) and gemcitabine monotherapy (28 days). Further increases in survival occurred in combination therapy groups BEZ235 + gemcitabine (30 days, P = 0.007), BEZ235 + EMAP (27 days, P = 0.02), gemcitabine + EMAP (31 days, P = 0.001), and BEZ235 + gemcitabine + EMAP (33 days, P = 0.004). BEZ235 has experimental PDAC antitumor activity in vitro and in vivo that is further enhanced by combination of gemcitabine and EMAP. These findings demonstrate advantages of combination therapy strategies targeting multiple pathways in pancreatic cancer treatment. J. Cell. Biochem. 113: 784–791, 2012. © 2011 Wiley Periodicals, Inc.

[1]  M. Lippman,et al.  Anti-vascular therapy: a new approach to cancer treatment. , 2000, The Western journal of medicine.

[2]  A. Gazdar,et al.  Dual phosphoinositide 3-kinase/mammalian target of rapamycin blockade is an effective radiosensitizing strategy for the treatment of non-small cell lung cancer harboring K-RAS mutations. , 2009, Cancer research.

[3]  Daniela Gabriel,et al.  Identification and characterization of NVP-BEZ235, a new orally available dual phosphatidylinositol 3-kinase/mammalian target of rapamycin inhibitor with potent in vivo antitumor activity , 2008, Molecular Cancer Therapeutics.

[4]  C. Sawyers,et al.  The phosphatidylinositol 3-Kinase–AKT pathway in human cancer , 2002, Nature Reviews Cancer.

[5]  M. Mackenzie.  Molecular therapy in pancreatic adenocarcinoma. , 2004, The Lancet. Oncology.

[6]  M. Schwarz,et al.  In vivo therapy of local tumor progression by targeting vascular endothelium with EMAP-II. , 2004, The Journal of surgical research.

[7]  Yixin Yao,et al.  The PI 3-kinase/Akt signaling pathway is activated due to aberrant Pten expression and targets transcription factors NF-κB and c-Myc in pancreatic cancer cells , 2004, Oncogene.

[8]  R. DePinho,et al.  Pancreatic cancer biology and genetics , 2002, Nature Reviews Cancer.

[9]  D. Ito,et al.  In vivo antitumor effect of the mTOR inhibitor CCI‐779 and gemcitabine in xenograft models of human pancreatic cancer , 2006, International journal of cancer.

[10]  P. Lollini,et al.  NVP-BEZ235 as a New Therapeutic Option for Sarcomas , 2010, Clinical Cancer Research.

[11]  N. Awasthi,et al.  Antitumor effects of EMAP II against pancreatic cancer through inhibition of fibronectin-dependent proliferation , 2010, Cancer biology & therapy.

[12]  R. Schwarz,et al.  An orthotopic in vivo model of human pancreatic cancer. , 1999, Surgery.

[13]  Raghu Kalluri,et al.  Fibroblasts in cancer , 2006, Nature Reviews Cancer.

[14]  E. Kohn,et al.  Endothelial monocyte activating polypeptide II induces endothelial cell apoptosis and may inhibit tumor angiogenesis. , 2000, Microvascular research.

[15]  M. Reni,et al.  Gemcitabine versus cisplatin, epirubicin, fluorouracil, and gemcitabine in advanced pancreatic cancer: a randomised controlled multicentre phase III trial. , 2005, The Lancet. Oncology.

[16]  D. Warburton,et al.  Endothelial Monocyte Activating Polypeptide II inhibits lung neovascularization and airway epithelial morphogenesis , 2000, Mechanisms of Development.

[17]  D. Hedley,et al.  Activity of a novel, dual PI3-kinase/mTor inhibitor NVP-BEZ235 against primary human pancreatic cancers grown as orthotopic xenografts , 2009, British Journal of Cancer.

[18]  R. Schmid,et al.  Pancreatic cancer: basic and clinical aspects. , 2005, Gastroenterology.

[19]  S. Corbett,et al.  Endothelial-monocyte activating polypeptide II alters fibronectin based endothelial cell adhesion and matrix assembly via alpha5 beta1 integrin. , 2005, Experimental cell research.

[20]  Huan Yang,et al.  The Akt/PKB pathway: molecular target for cancer drug discovery , 2005, Oncogene.

[21]  A. Abioye,et al.  Pancreatic carcinoma. , 2020, Journal of the National Medical Association.

[22]  N. Pellegata,et al.  K-ras and p53 gene mutations in pancreatic cancer: ductal and nonductal tumors progress through different genetic lesions. , 1994, Cancer research.

[23]  D. Bar-Sagi,et al.  Oncogenic K-ras Drives Cell Cycle Progression and Phenotypic Conversion of Primary Pancreatic Duct Epithelial Cells , 2004, Cancer Research.

[24]  M. Tsao,et al.  Wortmannin inhibits pkb/akt phosphorylation and promotes gemcitabine antitumor activity in orthotopic human pancreatic cancer xenografts in immunodeficient mice. , 2001, Clinical cancer research : an official journal of the American Association for Cancer Research.

[25]  A. Bergner,et al.  The dual PI3K/mTOR inhibitor BEZ235 is effective in lung cancer cell lines. , 2011, Anticancer research.

[26]  N. Awasthi,et al.  EMAP II-Based Antiangiogenic-Antiendothelial In Vivo Combination Therapy of Pancreatic Cancer , 2010, Annals of Surgical Oncology.

[27]  R. Andersson,et al.  Analysis of MUC4 expression in human pancreatic cancer xenografts in immunodeficient mice. , 2014, Anticancer research.

[28]  E. Dühmke,et al.  Combined chemoradiotherapy for isolated local recurrence after primary resection of pancreatic cancer. , 2005, JOP : Journal of the pancreas.

[29]  Jun Li,et al.  Endothelial-Monocyte Activating Polypeptide Ii, a Novel Antitumor Cytokine That Suppresses Primary and Metastatic Tumor Growth and Induces Apoptosis in Growing Endothelial Cells , 1999, The Journal of experimental medicine.

[30]  T. Khoury,et al.  Activated Akt and Erk Expression and Survival After Surgery in Pancreatic Carcinoma , 2006, Annals of Surgical Oncology.

[31]  N. Awasthi,et al.  Endothelial monocyte activating polypeptide II interferes with VEGF-induced proangiogenic signaling , 2009, Laboratory Investigation.

[32]  J. Baselga,et al.  NVP-BEZ235, a dual PI3K/mTOR inhibitor, prevents PI3K signaling and inhibits the growth of cancer cells with activating PI3K mutations. , 2008, Cancer research.

[33]  N. Awasthi,et al.  An antiendothelial combination therapy strategy to increase survival in experimental pancreatic cancer. , 2009, Surgery.

[34]  M. Tsao,et al.  Inhibition of phosphatidylinositide 3-kinase enhances gemcitabine-induced apoptosis in human pancreatic cancer cells. , 2000, Cancer research.

[35]  L. Tanoue.  Cancer Statistics, 2009 , 2010 .

[36]  K. Yamaguchi,et al.  Frequent expression of genes for receptor tyrosine kinases and their ligands in human pancreatic cancer cells , 1995, International journal of pancreatology : official journal of the International Association of Pancreatology.

[37]  M. Brennan.  Adjuvant therapy following resection for pancreatic adenocarcinoma. , 2004, Surgical oncology clinics of North America.

[38]  K. M. Chen,et al.  Dual Targeting of Phosphoinositide 3-Kinase and Mammalian Target of Rapamycin Using NVP-BEZ235 as a Novel Therapeutic Approach in Human Ovarian Carcinoma , 2011, Clinical Cancer Research.

[39]  H. Reber,et al.  Influence of hypoxia and neoangiogenesis on the growth of pancreatic cancer , 2003, Molecular Cancer.