8A) and measuring new particle production by titration of media on BHK-21 cells (Fig. levels of IFN-stimulated genes (ISGs) were resistant to apoptosis under most experimental conditions, even when VSV replication levels were dramatically increased by Jak inhibitor I treatment. Two of these cell lines also poorly activated apoptosis when treated with Fas activating antibody, suggesting a general defect in apoptosis. INTRODUCTION Oncolytic virus (OV) therapy is an innovative anticancer approach utilizing replication-competent viruses that preferentially infect and kill cancer cells [reviewed in (Russell et al., 2012)]. Vesicular stomatitis virus (VSV), a prototypic non-segmented negative-strand RNA virus (order em JNJ-40411813 Mononegavirales /em , family em Rhabdoviridae /em ), is a promising oncolytic virus against various malignancies [reviewed in (Barber, 2004; Hastie and Grdzelishvili, 2012)], and a phase I clinical trial using VSV against hepatocellular carcinoma is in progress (http://clinicaltrials.gov, trial “type”:”clinical-trial”,”attrs”:”text”:”NCT01628640″,”term_id”:”NCT01628640″NCT01628640). While wild type (wt) VSV cannot be utilized as an OV due to its unacceptable neurotoxicity, numerous VSV-based recombinants with significantly decreased neurotoxicity and improved oncoselectivity have been generated [reviewed in (Hastie and Grdzelishvili, 2012)]. One of the best performing oncolytic VSVs is VSV with replacement or deletion of the methionine at amino acid position 51 (M51) of the VSV matrix (M) protein. The oncoselectivity (and safety) of VSV M51 mutants is largely based on their inability to evade type I interferon (IFN) mediated antiviral responses in non-malignant cells (Ahmed et al., 2003; JNJ-40411813 Brown et al., 2009; Ebert O et al., 2005; Stojdl DF et al., 2003; Trottier et al., 2007; Wollmann G et al., 2010). However, cancer cells often have defects in type I IFN signaling, which can provide a growth advantage to uninfected cells, but impairs their ability to inhibit VSV infection and replication [reviewed in (Barber, 2005; Hastie et al., 2013; Lichty BD et al., 2004)]. Pancreatic cancer is one of the most lethal abdominal malignancies with annual deaths closely matching the annual incidence of the disease [reviewed in (Farrow B et al., 2008)]. About 95% of pancreatic cancers are pancreatic ductal adenocarcinomas (PDAC), which are highly invasive with aggressive local growth and rapid metastases to surrounding tissues [reviewed in (Stathis A and Moore, 2010)]. Our recent studies demonstrated that VSV is very effective against the majority of human PDAC cell lines, both in vitro and in vivo, but that some cell lines are resistant to VSV replication and oncolysis (Moerdyk-Schauwecker et al., 2013; Murphy et al., 2012). All cell lines resistant to VSV retained functional type I IFN responses (Moerdyk-Schauwecker et al., 2013; Murphy et al., 2012) and displayed constitutive high-level expression of JNJ-40411813 the IFN-stimulated antiviral genes MxA and OAS (Moerdyk-Schauwecker et al., 2013; Murphy et al., 2012)). Inhibition of JAK/STAT signaling by Jak inhibitor I (Jak Inh. I) decreased levels of MxA and OAS and JNJ-40411813 increased VSV replication (Moerdyk-Schauwecker et al., 2013). Effective oncolytic virus (OV) therapy depends not only on the ability of OVs to infect and replicate in cancer cells, but also to kill them. VSV kills infected cells primarily via induction of apoptosis (Balachandran et al., 2001; Balachandran et al., 2000; Cary et al., 2011; Gadaleta et al., 2005; Gaddy DF and and Lyles, 2005; Gaddy DF, 2007; Kopecky and Lyles, 2003; Kopecky et al., 2001). The specific mechanism of apoptosis CASP3 in response to VSV infection depends on both virus and cell type, and apoptosis induction has never been studied in any pancreatic cancer cells in response to JNJ-40411813 VSV. Thus, the goals of this study were (1) to investigate the mechanism of apoptosis induction in PDAC cell lines by three different viruses: wt-like VSV (VSV-GFP) and VSV attenuated by M dependent and independent mechanisms (VSV-M51-GFP and VSV-P1-GFP respectively; and (2) to examine whether dysregulation of apoptosis, a hallmark of PDACs as well as other cancers [reviewed in (Hamacher et al., 2008; Neesse et al., 2012; Roder et al., 2011)], contributes to the resistance of some PDACs to VSV-mediated oncolysis. For example, in chronic lymphocytic leukemia (CLL) cells overexpressing the anti-apoptotic protein Bcl-2, VSV-M51R (M51R substitution in M protein) was unable to induce apoptosis and consequently the CLL cells were resistant to VSV-induced killing (Tumilasci et al., 2008). The use of a VSV recombinant with the M51 deletion in the M protein (unable to evade type I IFN responses), and.