190 Development of an HLA A*0201 Restricted T Cell Receptor That Recognizes Peptide Epitopes from MAGE A3 and MAGE A12 for Targeted Adoptive T Cell Immunotherapy of Multiple Cancer Types Molecular The[.]
CANCER - IMMUNOTHERAPY I its aim, we attempted to present interleukin (IL)-12 on the surface of HVJ-E We constructed a mouse IgG constant region (Fc)-displayed HVJ-E without HN (Fc-DHN-HVJ-E) and produced a single-chain interleukin-12 (scIL12) fused with ZZ domain of Protein A (ZZscIL12) Fc-DHN-HVJ-E was connected with ZZ-scIL12 by binding Fc and ZZ domain, and it was possible to have present ZZ-scIL12 on the surface of Fc-DHN-HVJ-E ZZ-scIL12-presented Fc-DHNHVJ-E (scIL12-HVJ-E) could activate dendritic cells and splenocytes, and induced secretion of IFN-g from them Moreover, the anti-tumor effect of scIL12-HVJ-E was much more powerful than conventional HVJ-E in mice bearing F10 melanoma and complete eradication was achieved in approximately 30% of tumor-bearing mice by scIL12HVJ-E Systemic injection of scIL12-HVJ-E suppressed metastatic melanoma nodules in lung In this study, we succeeded in enhancing the anti-tumor effect of HVJ-E by means of the scIL12 presentation The high-performance HVJ-E will be available for cancer treatment by systemic administration in future 188 Combined Gene Therapy Using Adenovirus Expressing Interleukin-12, GranulocyteMacrophage Colony-Stimulating Factor, and Thymidine Kinase with Prodrug Ganciclovir Ji-seong Kim,1,2 Kyoung-ju Choi,1,2 Chae-Ok Yun.1,2 Brain Korea 21 Project for Medical Science, Yonsei University College of Medicine, Seoul, Republic of Korea; 2Brain Korea 21 Project for Medical Science, Yonsei University College of Medicine, Seoul, Republic of Korea Tumors exhibit immune escape properties by generating immunosuppressive tumor microenvironment and subverting host immune responses that nally promote their survival Accordingly, Immunotherapy has been investigated with various strategies involving multiple elements such as combination with several cytokines, co-stimulatory molecules and DC vaccination to improve immune system in tumor microenvironment Despite of these efforts, still cancer immunotherapy is limited by various huddles Especially, lack of tumor susceptibility is one of the focused problems To further investigate the efcient and synergistic cancer immunotherapy by improving tumor susceptibility and tumor cell killing, combined cytokine and “suicide gene” therapy can be novel approach that leads to stimulate tumor specic immune system In this study, we generated IL-12, GM-CSF and HSV-TK co-expressing oncolytic Ads for additive and synergistic anti-tumor immune responses and therapeutic effects through activating type-I immune response and inducing DC activation via enhancing tumor susceptibility In vitro assay, cell viability of RdB/IL12/GMCSF-TK Ads treated with ganciclovir remarkably decreased in viral and ganciclovir dosedependent manner Besides, combinational administration of RdB/ IL12/GMCSF-TK Ads with ganciclovir showed signicant antitumor effect and prolonged survival rate were observed compared with control virus in vivo Moreover, massive inltration of CD4+, CD8+ T cells into tumor tissues were observed and also enhanced tumorspecic CTL and increased numbers of IFN-gamma secreting T cells were induced in splenocytes of co-treated group Overall, our results indicate that combined IL-12, GMCSF and suicide gene therapy is a promising cancer immunotherapy by synergistic anti-tumor immune response S72 189 Safety Data of MGN1601, a Tumor Vaccine, Made of Allogeneic, Transfected and Irradiated Tumor Cells in Combination with an Immunomodulator for the Treatment of Metastatic Renal Cell Carcinoma Manuel Schmidt,1 Baerbel Volz,1 Matthias Schroff,1 Kerstin Kapp,1 Christiane Kleuss,1 Marina Tschaika,1 Burghardt Wittig.1,2 Mologen AG, Berlin, Germany; 2Charite Universitaetsmedizin Campus Benjamin Franklin, Institute foe Molecular Biology and Bioinformatics, Berlin, Germany Background The cell-based tumor vaccine MGN1601 for the treatment of renal cell carcinoma (RCC), consists of two active pharmaceutical ingredients: First, genetically modied allogeneic (human) cells in-vitro transiently transfected with four different MIDGE® vectors encoding IL-7, GM-CSF, CD80 and CD154 and second, the synthetic DNA-based immunomodulatory molecule dSLIM®-30L1, a TLR-9 agonist The cells originate from primary RCC material Methods Studies of single-dose toxicity were performed with MGN1601 in rats (heterologous setting) by either subcutaneous or intradermal application (up to 300-fold or 60-fold of the amount of cells for patients/kg, respectively) Additionally, a single dose subcutaneous application of the murine homologue of MGN1601 in NMRI mice was performed (allogeneic setting, up to 2.900-fold of the amount of cells for patients/kg) Studies of repeateddose toxicity (5- and 13-times, 28 days and 13 weeks, respectively) were performed in either the heterologous setting or the homologue setting Furthermore, the local tolerance of MGN1601 was tested in rabbits Results The vaccine generally showed low to no toxicity at the tested doses In all single-dose toxicity studies the NOEL was higher than the highest dose used In the repeated-dose 28 days toxicity studies (5-times) only slight hematological changes in some female rats from the high-dose group (decrease of neutrophils and increase of lymphocytes) as well as minor changes at the injection site in some animals of all dose groups were observed Conclusions The murine homologue of MGN1601 applied to mice as well as MGN1601 in the heterologoues setting in rats showed excellent safety characteristics and was extremely well tolerated in all tested models up to very high multiples of the anticipated human therapeutic dose/ kg A clinical trial Phase I/II was initiated in November 2009 First patients will be included in the 1st Q/2010 The treatment duration will be months An extension treatment will last for further years In this clinical study safety and efcacy of MGN1601 in patients with metastatic RCC will be evaluated 190 Development of an HLA-A*0201 Restricted T Cell Receptor That Recognizes Peptide Epitopes from MAGE-A3 and MAGE-A12 for Targeted Adoptive T Cell Immunotherapy of Multiple Cancer Types Nachimuthu Chinnasamy,1 Jennifer A Wargo,2 Zhiya Yu,1 Timothy L Frankel,1 John P Riley,1 Maria R Parkhurst,1 Steven A Feldman,1 Nicholas P Restifo,1 Paul F Robbins,1 Steven A Rosenberg,1 Richard A Morgan.1 Surgery Branch, National Cancer Institute, Bethesda, MD; Surgery Department, Massachusetts General Hospital, Boston, MA Adoptive cell transfer therapy using peripheral blood lymphocytes (PBL) genetically engineered to express tumor antigen-specic T cell receptors (TCRs) is a promising strategy for treating patients with malignancies TCR engineered PBL targeting melanocyte differentiation antigens mediated objective tumor regression in patients with metastatic melanoma However, this therapy has also been shown to lead to severe on-target toxicity against skin, eye and ear, normal tissue that contain antigen-expressing melanocytes Molecular Therapy Volume 18, Supplement 1, May 2010 Copyright © The American Society of Gene & Cell Therapy CANCER-ONCOLYTIC VIRUSES I (Johnson LA et al, Blood, 114: 535-546, 2009) In an attempt to avoid toxicity to normal tissue, TCRs were derived that recognize cancer testis antigens (CTA) These antigens are expressed in multiple tumors with no expression in normal tissue with the exception of immunoprivileged sites such as testis and placenta that lack expression of MHC class I molecules The MAGE family of CTA represent attractive candidates for targeted adoptive immunotherapy of cancer and one of the members of this family MAGE-A3 is expressed in >70% of metastatic melanomas, as well as a wide range of nonmelanoma epithelial malignancies including lung, ovarian, bladder, head and neck carcinomas, multiple myeloma and hepatocellular carcinoma Initially, transgenic mice that expresses the human class I HLA-A*0201 molecule were immunized with either of two candidate peptides, MAGE-A3: 112-120 (KVAELVHFL) or 271-279 (FLWGPRALV) with a helper peptide (HBVc: 128-140) Murine T cell clones were generated from the splenocytes following repeated in vitro stimulation with peptides and MAGE-A3 specic TCR α and β chains were then isolated, tested for reactivity, and cloned into an MSGV1 based retroviral vector Two TCRs recognizing the epitopes were selected for detailed evaluation Expression of both TCRs in human PBL demonstrated antigen-specic reactivity in the form of tumor cell lysis and cytokine secretion against a range of melanoma and non-melanoma tumor cell targets Based on the levels of antigen specic reactivity against target cells, the TCR against MAGE-A3: 112-120 was selected for further development Peptide epitopes from both MAGE-A3 and A12 were efciently recognized by the TCR engineered PBL, thus potentially broadening the number of tumors that can be targeted by this TCR In an effort to improve the function of the TCR, single amino acid substitution variants of the CDR3 region in the α chain were generated Substitution of alanine to threonine at position 118 in the CDR3 region of the α chain of MAGE-A3: 112-120 TCR improved its function Based on these results a clinical trial is planned in which PBL will be transduced with the optimized TCR and transferred to the autologous patient in an attempt to treat MAGE-A3 and/or MAGE-A12 expressing tumors of different histologies Cancer-Oncolytic Viruses I 191 A Comparative Study of Stem Cell-Based Cell Carrier Systems for the Systemic Delivery of an Oncolytic Adenovirus for Antiglioma Therapy Matthew A Tayler,1 Atique U Ahmed,1 Ilya V Ulasov,1 Irina Y Balyasnikova,1 Maciej S Lesniak.1 The University of Chicago Brain Tumor Center, The University of Chicago, Chicago, IL CRAd-S-pk7 is a survivin promoter regulated oncolytic adenovirus, which has shown optimistic anti-glioblastoma (GBM) activity in the preclinical setting However, successful application of this approach will require enhanced delivery of the therapeutic virus in order to treat disseminated tumor burden in the brain The discovery of the inherent tumor-tropic properties of the stem cells provides a unique opportunity for targeted therapy that employs stem cells as cellular vehicles to track metastatic tumor burden and also deliver the therapeutic payload at the tumor sites without activating the host immune system Based on this, we optimized the stem cell based delivery system by comparing human neural stem cell (NSC) and mesenchymal stem cell (MSC) as a cell carrier to deliver oncolytic adenovirus systemically for antiglioma therapy We rst evaluated the direct migratory capacity of the NSCs and the MSCs to glioma both in vitro and in vivo and observed that NSCs have a signicantly greater specic migratory activity in response to two primary GBM samples and glioma cell lines (P70% of metastatic melanomas,... will be transduced with the optimized TCR and transferred to the autologous patient in an attempt to treat MAGE- A3 and/ or MAGE- A12 expressing tumors of different histologies Cancer- Oncolytic Viruses... not associated with the entry of CVB into cancer cells Flow cytometric annexin V assay demonstrated that the main oncolytic effects of CVB for cancer cells were attributed not to apoptosis but