title: Insulin Receptor and GPCR Crosstalk Stimulates YAP via PI3K and PKD in Pancreatic Cancer Cells.
abstract: We examined the impact of crosstalk between the insulin receptor and G protein-coupled receptor (GPCR) signaling pathways on the regulation of Yes-associated protein (YAP) localization, phosphorylation, and transcriptional activity in the context of human pancreatic ductal adenocarcinoma (PDAC). Stimulation of PANC-1 or MiaPaCa-2 cells with insulin and neurotensin, a potent mitogenic combination of agonists for these cells, promoted striking YAP nuclear localization and decreased YAP phosphorylation at Ser127and Ser397Challenging PDAC cells with either insulin or neurotensin alone modestly induced the expression of YAP/TEAD-regulated genes, including connective tissue growth factor (CTGF), cysteine-rich angiogenic inducer 61 (CYR61), andCXCL5, whereas the combination of neurotensin and insulin induced a marked increase in the level of expression of these genes. In addition, siRNA-mediated knockdown of YAP/TAZ prevented the increase in the expression of these genes. A small-molecule inhibitor (A66), selective for the p110α subunit of PI3K, abrogated the increase in phosphatidylinositol 3,4,5-trisphosphate production and the expression ofCTGF, CYR61, andCXCL5induced by neurotensin and insulin. Furthermore, treatment of PDAC cells with protein kinase D (PKD) family inhibitors (CRT0066101 or kb NB 142-70) or with siRNAs targeting the PKD family prevented the increase of CTGF, CYR61, and CXCL5 mRNA levels in response to insulin and neurotensin stimulation. Thus, PI3K and PKD mediate YAP activation in response to insulin and neurotensin in pancreatic cancer cells.Implications:Inhibitors of PI3K or PKD disrupt crosstalk between insulin receptor and GPCR signaling systems by blocking YAP/TEAD-regulated gene expression in pancreatic cancer cells.Mol Cancer Res; 15(7); 929-41. ©2017 AACR.
authors.affiliation: Tianjin Medical University, Tianjin, China.|Division of Digestive Disease, Department of Medicine, David Geffen School of Medicine, Los Angeles, California.
authors.forename: Fang
authors.initials: F
authors.lastname: Hao
authors.affiliation: Division of Digestive Disease, Department of Medicine, David Geffen School of Medicine, Los Angeles, California.|Xi'an Jiaotong University, Xi'an, China.
authors.forename: Qinhong
authors.initials: Q
authors.lastname: Xu
authors.affiliation: Division of Digestive Disease, Department of Medicine, David Geffen School of Medicine, Los Angeles, California.|Sichuan University, Chengdu, China.
authors.forename: Yinglan
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Division of Digestive Disease, Department of Medicine, David Geffen School of Medicine, Los Angeles, California.
authors.forename: Jan V
authors.initials: JV
authors.lastname: Stevens
authors.affiliation: Division of Digestive Disease, Department of Medicine, David Geffen School of Medicine, Los Angeles, California.|CURE: Digestive Disease Research Center, Los Angeles, California.|VA Greater Los Angeles Health Care System, Los Angeles, California.
authors.forename: Steven H
authors.initials: SH
authors.lastname: Young
authors.affiliation: Division of Digestive Disease, Department of Medicine, David Geffen School of Medicine, Los Angeles, California.|CURE: Digestive Disease Research Center, Los Angeles, California.|VA Greater Los Angeles Health Care System, Los Angeles, California.
authors.forename: James
authors.initials: J
authors.lastname: Sinnett-Smith
authors.affiliation: Division of Digestive Disease, Department of Medicine, David Geffen School of Medicine, Los Angeles, California. erozengurt@mednet.ucla.edu.|CURE: Digestive Disease Research Center, Los Angeles, California.|VA Greater Los Angeles Health Care System, Los Angeles, California.|Molecular Biology Institute, University of California at Los Angeles, Los Angeles, California.
authors.forename: Enrique
authors.initials: E
authors.lastname: Rozengurt
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics*; D015703:Antigens, CD / Q000235:genetics; D021441:Carcinoma, Pancreatic Ductal / Q000188:drug therapy / Q000235:genetics* / Q000473:pathology; D045744:Cell Line, Tumor; D054365:Chemokine CXCL5 / Q000235:genetics; D058534:Class I Phosphatidylinositol 3-Kinases / Q000235:genetics*; D055513:Connective Tissue Growth Factor / Q000235:genetics; D055515:Cysteine-Rich Protein 61 / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D007328:Insulin / Q000008:administration & dosage / Q000378:metabolism; D009496:Neurotensin / Q000008:administration & dosage / Q000378:metabolism; D010190:Pancreatic Neoplasms / Q000188:drug therapy / Q000235:genetics* / Q000473:pathology; D010750:Phosphoproteins / Q000235:genetics*; D010766:Phosphorylation / Q000187:drug effects; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000235:genetics*; D011743:Pyrimidines / Q000008:administration & dosage; D011972:Receptor, Insulin / Q000235:genetics; D043562:Receptors, G-Protein-Coupled / Q000235:genetics; D014157:Transcription Factors; D000091102:YAP-Signaling Proteins
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D015703:Antigens, CD; C501848:CCN1 protein, human; C526317:CCN2 protein, human; C551536:CRT 0066101; C486263:CXCL5 protein, human; D054365:Chemokine CXCL5; D055515:Cysteine-Rich Protein 61; D007328:Insulin; D010750:Phosphoproteins; D011743:Pyrimidines; D043562:Receptors, G-Protein-Coupled; D014157:Transcription Factors; D000091102:YAP-Signaling Proteins; C088374:YAP1 protein, human; D055513:Connective Tissue Growth Factor; D009496:Neurotensin; D058534:Class I Phosphatidylinositol 3-Kinases; C484760:PIK3CA protein, human; C089212:protein kinase D; C435941:INSR protein, human; D011972:Receptor, Insulin; D011493:Protein Kinase C