680 Gene Therapy of Chondroitin Sulfate Coated Ternary GM CSF Plasmid Complex for Ovarian Cancer Molecular Therapy Volume 20, Supplement 1, May 2012 Copyright © The American Society of Gene & Cell The[.]
CANCER – IMMUNOTHERAPY III important therapeutic functions, such as cytokine production, cellular phenotype, and antigen processing capabilities, are tested in vitro and compared to unlabeled control In vivo results demonstrate the ability of 19F MRI to detect and monitor the location of the cells postinjection Since there is little to no native fluorine in biological tissues, any 19F signal exceeding noise threshold can be directly attributed to the cellular label Using conventional MRI, the anatomical position of the cells can be precisely determined in three dimensions, yielding the ability to calculate a quantitative measure of therapeutic cell density in known anatomical location Using contralateral control as well as DC’s dual-labeled with luciferase, details of contrast agent effects on cell viability in vivo, the anatomical location of cell death, and migration to the draining lymph node are examined This dataset in its entirety provides persuasive evidence of the safety and effectiveness of cell tracking in a clinical setting in which patients with advanced CRC showing disease control after first-line therapy with standard chemotherapy regimen are included The study treatment (60 mg MGN1703 or placebo) is administered subcutaneously twice weekly in a ratio 2:1 The therapy will be continued until tumor progression, intolerable toxicity, exclusion criteria or withdrawal of consent The study is conducted in Germany, Austria, France, UK, Czech Republic and Russia, and 129 patients have to be recruited into the study The efficacy and safety of the study treatment will be evaluated based on extensive immunological tests, radiological assessment, safety laboratory results and assessments of the quality of life Results and discussions: The patients received up to 132 treatment administrations (mean: 32.1) The majority of adverse events (84%) reported in the study so far have been assessed as not drug-related by the investigator The remaining AEs (16%) include mild fever, injection site itching, muscle aching, arthralgia, fatigue, paresthesia, rash, and moderate subfebril temperature and increased ANA in single patients Currently, nine serious adverse events (SAEs) have been reported of which only one was assessed as probably drug-related (“atypical pneumonia”) Local reactions reported in single patients include such symptoms as mild redness and swelling at injection site No laboratory or clinical signs of doselimiting toxicities have been reported, so far Conclusions: These preliminary safety results of the ongoing clinical study in patients with advanced CRC show that treatment with MGN1703 at the dosage of 60 mg is well tolerated and safe Reported AEs which were assessed as possibly drug-related belong to expected study drug reactions known for immune modulating drugs 680 Gene Therapy of Chondroitin SulfateCoated Ternary GM-CSF Plasmid Complex for Ovarian Cancer Katsuyuki Hamada,1 Chieko Yoshihara,2 Tomoko Ito,3 Yoshiyuki Koyama,2 Hiroshi Ito,3 Katsuko Takagi,1 Akihiro Nawa.1 Obstetrics and Gynecology, Ehime University, Toon, Ehime, Japan; 2Textile Science, Otsuma Women’s University, Chiyoda–ku, Tokyo, Japan; 3Pharmaceutical Sciences, Musashino University, Nishi, Tokyo, Japan; 4Animal Medical Center, Tokyo University of Agriculture and Technology, Fuchu, Tokyo, Japan Figure 19F MRI of DC migration to draining lymph nodes after foot-pad injection in a mouse model 679 Preliminary Safety Results from the IMPACT Study – A Phase 2-3 Clinical Study in Patients with Advanced Colorectal Carcincoma with the Immunomodulator MGN1703 Manuel Schmidt,1 Hans-Joachim Schmoll,2 Jorge RieraKnorrenschild,3 Avgust Garin,4 Mikhail Byakhov,5 Gunter Schuch,6 Dieter Nitsche,7 Ekaterina Weith,1 Matthias Schroff,1 Maxim Krikov,1 Marina Tschaika,1 Burghardt Wittig.8 Mologen AG, Berlin, Germany; 2University Clinic Halle (Saale), Halle, Germany; 3Universitätsklinikum Giessen und Marburg, Marburg, Germany; 4Russian Scientific Oncology Center RAMN, Moscow, Russian Federation; 5Central Clinical Hospital No RZHD, Moscow, Russian Federation; 6Hämatologischeonkologische Praxis Altona, Hamburg, Germany; 7Barmherzige Schwestern Linz, Linz, Austria; 8Foundation Institute Molecular Biology and Bioinformatics, Freie Universität Berlin, Germany Introduction: A phase 2-3 study was initiated in patients with advanced CRC having disease control after first-line therapy using the synthetic DNA-based immunomodulator MGN1703 which acts as an agonist of toll-like receptor The objective of the study is to assess efficacy and safety of the MGN1703 treatment in comparison to placebo Material and methods: The IMPACT study is designed as a randomized double-blind placebo-controlled phase 2-3 study, S262 Although replication-competent viruses have been developed to treat cancers, their cytotoxic effects are insufficient due to infection inhibited by generation of neutralizing antibodies and systemic administration is difficult due to life-threatening serious side effects of virus-induced cytokine surge To overcome these critical problems, we devised a plasmid/polycation/polyanion complex and assessed the potential of ternary plasmid complexes coated with chondroitin sulfate in gene therapy for ovarian cancer The antitumor effects of chondroitin sulfate-coated complex as an anionic component were compared to those of hyaluronic acid on ovarian cancer Plasmid harboring the gene of murine granulocyte macrophage-colony stimulating factor (mGM-CSF) was complexed with polyethyleneimine (PEI) and hyaluronic acid or chondroitin sulfate Murine OVHM cells were injected into (C57BL/6 × C3H/He) F1 mice to prepare a subcutaneous or intraperitoneal tumor model DNA/PEI was charged positively and DNA/PEI/chondroitin sulfate or DNA/PEI/hyaluronic acid was charged negatively Plasmid-green fluorescent protein (GFP)/PEI coated with 10-kilodalton chondroitin sulfate increased transfection efficiency compared to coating with chondroitin sulfate of higher-molecular-weight or hyaluronic acid The transfection efficiency of GFP/PEI/10-kilodalton chondroitin sulfate in ovarian cancer cells was times that in normal cells Intraperitoneal injection of mGM-CSF/PEI coated with 10-kilodalton chondroitin sulfate prolonged survival compared to that coated with hyaluronic acid Intratumoral injection of mGM-CSF/PEI coated with 10-kilodalton chondroitin sulfate achieved 100% rates of mouse Molecular Therapy Volume 20, Supplement 1, May 2012 Copyright © The American Society of Gene & Cell Therapy CANCER-TARGETED GENE & CELL THERAPY: VECTOR BASED TECHNOLOGY survival but that with hyaluronic acid did not These findings suggest that GM-CSF/PEI coated with 10-kilodalton chondroitin sulfate has potential for use in gene therapy of ovarian cancer 681 Syngeneic Hematopoietic Stem Cell Transplantation Enhances the Antitumor Immunity of Intratumoral Type I Interferon Gene Transfer for Sarcoma Kenta Narumi,1 Takeshi Udagawa,1 Koji Suzuki,1 Kouichirou Aida,1 Atsushi Makimoto,2 Kazunori Aoki.1 Division of Gene and Immune Medicine, National Cancer Center Research Institute, Tokyo, Japan; 2Division of Pediatric Oncology, National Cancer Center Hospital, Tokyo, Japan Sarcoma at advanced stages remains a clinically challenging disease, and new therapeutic approaches are needed in the therapeutic strategy of malignant bone and soft tissue sarcoma Interferons (IFN) can target cancer cells by multiple antitumor activities including the induction of cancer cell death and enhancement of immune response However, the development of an effective cancer immunotherapy is often difficult because cancer generates an immunotolerant microenvironment against the host immune system Hematopoietic stem cell transplantation (HSCT) following a preconditioning is expected to introduce a fresh immune system, in which tolerance to tumor cells is not yet induced, and may present an opportunity to augment the efficacy of IFN immune therapy Recently, we found that the frequency of Foxp3+ cells per CD4+ cells is increased in the spleens, whereas the frequency is clearly decreased in the tumors after HSCT Furthermore, it has been reported recently that autologous HSCT can induce antitumor immunity by homeostatic proliferation (HP) of T cells following preconditioning-induced lymphopenia Here we examined whether a combination of autologous HSCT and IFN could induce an effective tumor-specific immune response against sarcoma First, we found that a type I IFN gene transfer significantly suppressed the cell growth of various sarcoma cell lines, and that IFN-β gene transfer was more effective in inducing cell death than was IFN-α in sarcoma cells Then, to examine the antitumor effect in vivo, human sarcoma cells were inoculated in immune-deficient mice, and a lipofection of an IFN-β-expressing plasmid was found to suppress the growth of subcutaneous tumors significantly Finally, the IFN gene transfer was combined with syngeneic HSCT in murine osteosarcoma models Intratumoral IFN-β gene transfer markedly suppressed the growth of vector-injected tumors and inhibited formation of spontaneous lung and liver metastases in syngeneic HSCT mice, and an infiltration of many CD4+ and CD8+ cells was recognized in metastatic tumors of the treated mice The treated mice showed no significant adverse events A combination of intratumoral IFN gene transfer with autologous HSCT could be a promising therapeutic strategy for patients with sarcoma This therapeutic strategy deserves an evaluation in future clinical trials for patients with sarcomas 682 A Protocol for Ex Vivo Expansion of Effector T Cells That Are Resistant to Oxidative and Apoptotic Stress Chuan Jin,1 Di Yu,1 Victoria Rashkova,1 Berith Nilsson,1 Fredrik Eriksson,1 Alex Karlsson-parra,1 Magnus Essand.1 Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden Adoptive transfer of tumor infiltrating lymphocytes or genetically engineered T cells are emerging cancer therapeutics Such cells need to be expanded ex vivo before being infused Commonly used protocols for expansion are either to use anti-CD3/CD28 mAb-coated magnetic beads or to use an agonistic CD3 antibody (OKT 3), in combination with IL-2 and, sometimes, irradiated allogeneic PBMCs as feeder cells This leads to efficient proliferation, possibly at the Molecular Therapy Volume 20, Supplement 1, May 2012 Copyright © The American Society of Gene & Cell Therapy cost of loss of quality since the expanded T cells are often short-lived once adoptively transferred back to the patient Naïve CD8+ T cells needs to be activated by mature dendritic cells (DCs) to become cytolytic killer T cells The maturation status, quality and cytokine profile of the DC is crucial for the outcome of the T cell One of the strongest sensitizers of DCs is to expose them to allogeneic cells We hypothesized, that stronger T cells could be obtained if they are stimulated with a mixture of dendritic cells (DCs) and allo-sensitized allogeneic lymphocytes Such cell products can be produced at large quantities and stored in advance When comparing T cells stimulated this way with T cells expanded with the rapid expansion protocol (OKT 3, IL-2 and irradiated PBMCs) we found that the T cells expanded equally well and kill target cells equally well but the DC/ allo-sensitized allogeneic lymphocytes stimulated T cells were much more resistant to the exposure of H2O2, immunosuppressive cytokines (IL-10 and TGFβ1) and apoptosis inducer We therefore propose this protocol for the ex vivo expansion of T cells Cancer-Targeted Gene & Cell Therapy: Vector Based Technology 683 Exploiting Endogenous microRNA Regulation To Target Gene Therapy to Tumors by Infiltrating Macrophages Roberta Mazzieri,1 Giulia Escobar,1 Davide Moi,1 Anna Ranchetti,1 Michele De Palma,1 Luigi Naldini.1 Angiogenesis and Tumor Targeting Research Unit, San Raffaele Telethon Institute for Gene Therapy (HSR-TIGET), Milan, Italy TEMs (Tie2-expressing monocytes), are recruited to tumors, where they promote angiogenesis and tumor growth We previously exploited the tumor homing ability and selective expression of the angiopoietin TIE2 receptor of TEMs to turn these cells into IFN-α cell vehicles, by engineering hematopoietic stem/progenitor cells (HSPCs) to express an IFN-α transgene only in TEM progeny This cell-based strategy targeted the IFN response to tumors, achieving antitumor activity in mice We now aim to increase the safety and feasibility of the strategy toward the goal of a clinical translation Being TIE2 also expressed by primitive HSPCs, we have developed new vectors with de-targeted transgene expression from HSPCs via post-transcriptional regulation mediated by microRNA We are currently establishing the safety of IFN-α delivery by the novel vectors monitoring HSC status and long-term repopulating activity in murine and human hematochimeric mice transplanted with gene modified HSPCs, and validating efficacy in relevant tumor models In mice, detargeting of the transgene expression from HSPCs did not affect the ability of TEM-mediated IFN-α delivery to inhibit both primary and secondary spontaneous mammary tumors Concerning the specificity of the human TIE2 promoter, FACS analysis on peripheral blood of human hematochimeric NSG mice showed, as expected, induced GFP expression in the myeloid compartment Moreover, human Tie2-GFP myeloid cells were recruited to human tumors orthotopically injected S263 ... potential for use in gene therapy of ovarian cancer 681 Syngeneic Hematopoietic Stem Cell Transplantation Enhances the Antitumor Immunity of Intratumoral Type I Interferon Gene Transfer for Sarcoma.. .CANCER- TARGETED GENE & CELL THERAPY: VECTOR BASED TECHNOLOGY survival but that with hyaluronic acid did not These findings suggest that GM- CSF/ PEI coated with 10-kilodalton chondroitin sulfate. .. maturation status, quality and cytokine profile of the DC is crucial for the outcome of the T cell One of the strongest sensitizers of DCs is to expose them to allogeneic cells We hypothesized, that