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　orMiaPaCa-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　Ser(127)　and　Ser(397).　Challenging　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),　and　CXCL5,　whereas　the　combination　of　neurotensin　and　insulin　induced　a　marked　increase　in　the　level　of　expression　of　these　genes.　In　addition,　siRNAmediated　knockdown　of　YAP/TAZ　prevented　the　increase　in　the　expression　of　these　genes.　A　small-molecule　inhibitor　(A66),　selective　for　the　p110　alpha　subunit　of　PI3K,　abrogated　the　increase　in　phosphatidylinositol　3,4,5-trisphosphate　production　and　the　expression　of　CTGF,　CYR61,　and　CXCL5　induced　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.