180 induction of anti tumor immunity by the baculovirus autographa californica multiple nuclear polyhedrosis virus

180 Induction of Anti Tumor Immunity by the Baculovirus Autographa Californica Multiple Nuclear Polyhedrosis Virus 178 Spontaneous Glioblastoma in Dogs Preclinical Model for High Capacity Adenoviral V[.]

178 Spontaneous Glioblastoma in Dogs: Preclinical Model for High Capacity Adenoviral Vector-Mediated Experimental Gene Therapy M Candolfi,' J Curtin,' W S Nichols,' A K M O Muham- 179 A Phase I Clinical Trial of LiposomeMediated Interferon-beta Gene Therapy for HighGrade Gliomas Toshihiko Wakabayashi, I Masaaki Mizuno,' Atsushi Natsume,' M O Castro.' 'Center or Genetic and Regenerative Medicine Nagoya University Hospital Nagoya Aichi, Japan ; "Neurosurgely, Nagoya University School ofMedicine, Nagoya Aichi, Japan mad,' M Puntel,' O D King,' W Xiong,' K M Kroeger,1 C Liu,' O E Pluhar,' E A Mc Nicl,2 r, R Ohlfcst,' A B Freese,' P F Moore,' D Palmer;" P Ng,6 J D Young," P R Lowenstein,' 'Gene Therapeutics Research Institute, Cedars-Sinai Medical Center-UCLA, Los Angeles, CA; 2Dept ofPathology and LaboratoryMedicine, Cedars-Sinai Medical Center; Los Angeles CA; 'Dept of Veterinary Clinical Sciences University ofMinnesota, Saint Paul MN; 4DepII of Neurosurgery, University ofMinnesota, Saint Paul, MN; :;Pathology Microbiology & Immunology School of Veterinary Medicine University ofCalifornia Davis, CA; "Dept ofMolecular and Human Genetics Baylor College ofMedicine, Houston, T){; 'Dept ofComparative Medicine Cedars-Sinai Medical Center; Los Angeles, CA Wcevaluated the dog spontaneous GBM as a large animal model for translational gene therapy strategies We determined the neuropathology of these tumors and the extent to which it resembles human OBM We also determined the feasibility of infecting dog normal brain tissue by injectingadenoviral vectors(Ads) expressing reporter and therapeutic transgenes in the cortex of healthy Beagle dogs Since human patients usually exhibit preexisting immunity against Ads, we determined the anti-Ad immune status of outbreed dogs Wefound that dog OBMs exhibit characteristic features of the humanGBM, includingnecrosis,pseudopalizading, neovascularization,endothelial proliferationand invasionintonon-neoplasticbrain In the brain of healthy Beagle dogs we administered Ad expressing the ~-Oalactosidase , or the therapeutic transgenes: TK, which kills glioma cells in the presenceofganciclovir (GCV), and the immunestimulatoryFIt3L, which attractsantigen presentingcells to the brain and the tumor mass Transgene expression was detected in neurons and astrocytes days post-injection without adverse clinical or neuropathologicalside effects.Weassessed the presenceofneutralizing antibodies in serum from 17 outbred dogs recruitedfor our study.We found that 60% exhibited neutralizingantibodies againstAds These data suggest that OBM-bearing dogs would also mimic the immune status of human patients in clinical trials Our results suggest that high capacity-Ads are excellent candidates for treating dog OBM, since their expression is not inhibitedby anti-Ad immune responses Thus, we assessed the ability ofHC-Ad vectors expressing TK and Flt3L to infect dog OBM cells in culture HC-Ads were effective at transducing dog GBM cells HC-Ad-TK elicited strong cytotoxic effects when combined with OCV.Our data indicate that dogs bearing spontaneous OBM constitute an attractive animal model for testing novel therapeutic approaches in a spontaneous tumor in the context of a larger brain Supported by: NIHININDS Grants IROI NS44556.01 MinoritySllpplementNS445561; I R21 NS054143-01 and I R03 TW006273-01 to M.G.C.; NIHININDSGrallfs I ROI NS 42893.01; U54 NS045309-01, and I R2 I NS047298-01 to PRL; the Bram and Elaine Goldsmith and the Medallions Group Endowed Chairs in Gene Therapeutics to PRL and MGC respectively and The Linda Tallen & David Paul Kane Foundation and the Board ofGovernors at CSMC S68 Masazumi Fujii,"Jun Yoshida." High-grade glioms are highly lethal neoplasms that represent about 20% of all intracranial tumors Cationic liposome-rnediated interferon-beta (IFN-~) gene transfer has been found to induce regression ofcxperimental glioma Weperformed a pilot clinical trial to evaluatesafetyand effectivenessofthis IFN-~ gene therapyin five patients with high-grade glioma Two patients showed more than 50% reductionand others had stabledisease 10weeks after the treatment initiation [Genetic analysis] In order to identify alterations in gene expression in brain tumors weeks after gene therapy trial, we uscd microarray technology and MetaCore analysis Interestingly, using hierarchical clustering and principal component analysis, series of gene therapy trial were classified according to response to I FNgene therapy There were significantchanges ingene expression which is related to immunoresponse and apoptosis, It confirms the validity of the methods Moreover, novel patterns of altered gene expression,such as inhibition of'neo-vascularization were identified, suggesting the involvement of pathways not previously described as involved [Brain autopsy] Autopsy revealed that tumor tissues showed dramatic changes after therapy in all patients Many tumor cells showednecroticchanges, and immunohistochemistry identified many CD8-positive lymphocytes and macrophages infiltrating in tumor and surrounding tissues, probably resulting from therapeutic effect Simultaneously, numberofMIB-1 positivecells was notable decreased No adverse findings associated with the clinical trial were pathologically observed [Conclusions] This study suggests gene therapy, which may become the feasibilityand safety ofIFN-~ an important treatment options for patients with malignant glioma Additional clinical trials are warranted 180 Induction of Anti-Tumor Immunity by the Baculovirus Autographa Californica Multiple Nuclear Polyhedrosis Virus Masayuki Kitajima, ' ,4 TakayukiAbc, I.' Naoko Miyano-Kurosaki,'·2 Toshinori Nakayama,4 Hiroshi Takaku.P I Department ofLife and Environmental Science, Chiba Institute ofTechnology; Narashino, Chiba , Japan ; "Hig h Technology Research, Chiba Institute ofTeelmology Narashino, Chiba, Japan; JResearch Centerfor Emerging lrfectious Diseases, Osaka University, Suita, Osaka Japan; 'Department ofImmunology, Graduate School ofMedicine Chiba University Clwo-ku Japan cnu« Background Wild type Autographa californica multiple nuclear polyhedrosis virus (AcMNPV) can infect a variety of mammalian cell types in vitro, but it does not replicate in these cells AcMNPV induces anti-viral cytokine production after injection in vivo, resulting in the protection of cells from infection with vesicular stomatitis virus and influenza virus Objective We here show that AcMNPV activates immune cells via TLR9/MyD88 signal pathway We also described the effects of AcMNPV on the induction of immune responsiveness and tumor growth in mice Results and Conclusions First, we found that AcNPV has the ability to induce innate immune system activation through a MyD88ffLR9-dependent pathway Next, we examined the effects ofAcMNPV in mice on immune responsiveness in general, and anti-tumor immunity in particular AeMNPV treatment also increased IFN-y levels in the serum and induced a marked elevation in IFN-y production Molecular Therapy Volume 15.Supplement tã.\by 2007 Copyright â The American $