- Email: [email protected]
187. Randomized Phase II Trial of Post-Operative Adjuvant Chemotherapy ±FANG™ Autologous Tumor Cell Vaccine in Colorectal Carcinoma with Liver Metastases
CANCER – IMMUNOTHERAPY I at 1.7 ± 0.2 months after gene therapy. Efficient micro-dystrophin expression was observed in the myocardium in treated mice. However, myocardial fibrosis was not mitigated. Most hemodynamic parameters were not improved either. Interestingly, treatment improved PR interval and QRS duration. Further, treated mice became more resistant to dobutamine-induced cardiac death. In summary, we have revealed for the first time the potential benefits and limitations of AAV micro-dystrophin therapy in end-stage Duchenne cardiomyopathy. Our findings have important implications on the further development of AAV micro-dystrophin gene therapy.
186. Myogenic Transduction and Cell Surface Marker Selection of DMD Fibroblasts Enable Stable Dystrophin mRNA Expression for Exon Skipping Assay
Takashi Saito,1,2 Tetsuya Nagata,1 Yoshitsugu Aoki,1 Jun Tanihata,1 Satoru Masuda,1 Toshifumi Yokota,3 Yuko Shimizu Motohashi,1 Shin’ichi Takeda.1 1 Department of Molecular Therapy, National Institute of Neuroscience, National Center of Neurology and Psychiatry, Kodaira, Tokyo, Japan; 2Department of Pediatrics, School of Medicine, Tokyo Women’s Medical University, Shinjuku, Tokyo, Japan; 3Department of Medical Genetics, University of Alberta Faculty of Medicine and Dentistry, Edmonton, AB, Canada.
Duchenne Muscular Dystrophy (DMD) is a lethal, X-linked muscular disease, due to the loss of dystrophin protein. Antisense oligonucleotide (AON) mediated exon skipping is one of promising approaches to cure this disease and phase II-III clinical trials of exon 51 skipping are ongoing. Preliminary but successful results of these trials are indicating that other exons will be targeted in subsequent trials. To develop effective AON for each DMD case, assays using patient-derived cells are valuable. In some cases, obtaining myoblasts from patients is difficult, therefore utilizing of fibroblasts is considered. We have previously reported the FACS-utilizing MyoD transduction of fibroblasts, and it was practical for exon skipping assay (PLoS One. 2010 Aug 18;5(8):e12239.). Nevertheless, in our current assay system, a long-term differentiation culture is required for enough dystrophin mRNA expression. To address this challenge, we evaluated an availability of several myogenic surface markers. CD56 is known as one of the myogenic markers expressed on a surface of primary myoblasts. We found that the MyoD-transduced fibroblasts, which were initially negative with CD56 by FACS analysis, gradually increased its positive population, and these cells expressed much dystrophin mRNA than negative population. These findings suggest that the expression of myogenic marker protein on MyoD-transduced fibroblasts would be beneficial to efficiently select the cells expressing dystrophin mRNA and shorten the period required for enough dystrophin mRNA expression. We would report the possibility that a stable dystrophin mRNA expression of DMD fibroblasts required for exon skipping assay might be achieved through a combination of MyoD transduction and myogenic surface marker.
Cancer – Immunotherapy I 187. Randomized Phase II Trial of PostOperative Adjuvant Chemotherapy ±FANG™ Autologous Tumor Cell Vaccine in Colorectal Carcinoma with Liver Metastases
Joseph Kuhn,5 Neil Senzer,1,2,3,4 Minal Barve,3 Padmasini Kumar,4 Donald D. Rao,4 Gladice Wallraven,4 Beena O. Pappen,4 Phillip B. Maples,4 John Nemunaitis.1,2,3,4 1 Mary Crowley Cancer Research Centers, Dallas; 2Medical City HCA, Dallas; 3Texas Oncology, P.A., Dallas; 4Gradalis, Inc., Dallas; 5WLS Surgical Associates, P.A., Dallas. The Phase I trial of FANG™ vaccine (comprised of irradiated autologous tumor cells transduced with plasmid encoding GMCSF and the RNA interference bi-shRNA FURIN; see Senzer et al., Mol Ther 2011) involving 31 previously treated solid tumor patients provided sufficient safety data to proceed to Phase II efficacy studies. This decision was bolstered by the correlation of survival with induced immune response (IFNgamma ELISPOT) in serially monitored patients at ≥ 4 months (p=0.025). Specifically, 6 metastatic refractory colon cancer patients in the Phase I trial, who having failed standard therapy, had an expected survival of ∼6 months, received a combined total of 24 vaccinations (i.e. 2 patients at dose level 1 x 107 and 4 patients at 2.5 x 107 cells / injection; data combined as there is no dose related difference in endpoint). These patients currently demonstrate median survivals of 358 days (554, 515+, 413, 303, 185, and 167 days each). Efficacy of the pbi-shRNA™ FURIN vaccine component was confirmed by marked knockdown of the furin proprotein convertase activated immunosuppressive proteins TGFβ1 and b2; 87% and 92% respectively). Based on these results, the Phase II trial (CL-PTL 114, BB-IND 14205) was activated following FDA, RAC, IBC and IRB acceptance. This is a randomized Phase II study of integrated adjuvant modified FOLFOX6 (mFOLFOX6) and intradermal autologous FANG™ cancer vaccine (1.0 x 107 cells/injection; maximum of 12 vaccinations) versus adjuvant mFOLFOX6 and placebo in patients with colorectal carcinoma with either synchronous or metachronous liver metastases (CLM ± pulmonary metastases) following resection ± ablation with curative intent. Eight weeks following resection ± ablation, patients will receive FANG™ Weeks 1, 3 then q 4 weeks with mFOLFOX starting Week 4 then continuing q 2 weeks x 6 months. A minimum harvest aliquot to produce 5 monthly injections will be required for entry into the study. These patients will be managed in an outpatient setting. Hematologic function, liver enzymes, renal function and electrolytes will be monitored weekly. Immune function analyses including assessment of tumor infiltrating lymphocytes (TIL) and ELISPOT analysis of cytotoxic T cell function to autologous tumor antigens will be monitored at baseline (TIL + ELISPOT) and thereafter ELISPOT analysis at Months 2, 4, 6, 9, end of treatment, and at response follow up visits. Preliminary data will be presented at the meeting.
188. Hematopoietic Stem Cell Immunotherapy for Melanoma
Eric H. Gschweng,1 Michael L. Kaufman,1 Roger P. Hollis,1 Koya C. Richard,2 Ribas Antoni,3 Donald B. Kohn.1 1 Molecular Biology, Immunology, and Molecular Genetics, UCLA, Los Angeles, CA; 2Department of Surgery, Surgical Oncology, UCLA, Los Angeles, CA; 3Department of Medicine, Hematology/ Oncology, UCLA, Los Angeles, CA. Melanoma is the deadliest form of skin cancer, and the primary treatment option of surgical removal becomes infeasible with widespread metastasis. Immunotherapy involves redirecting the immune system with transgenic T-cell receptors (TCR), and offers an attractive strategy to combat this disease. While transduction S74
Molecular Therapy Volume 20, Supplement 1, May 2012 Copyright © The American Society of Gene & Cell Therapy
186. Myogenic Transduction and Cell Surface Marker Selection of DMD Fibroblasts Enable Stable Dystrophin mRNA Expression for Exon Skipping Assay
Takashi Saito,1,2 Tetsuya Nagata,1 Yoshitsugu Aoki,1 Jun Tanihata,1 Satoru Masuda,1 Toshifumi Yokota,3 Yuko Shimizu Motohashi,1 Shin’ichi Takeda.1 1 Department of Molecular Therapy, National Institute of Neuroscience, National Center of Neurology and Psychiatry, Kodaira, Tokyo, Japan; 2Department of Pediatrics, School of Medicine, Tokyo Women’s Medical University, Shinjuku, Tokyo, Japan; 3Department of Medical Genetics, University of Alberta Faculty of Medicine and Dentistry, Edmonton, AB, Canada.
Duchenne Muscular Dystrophy (DMD) is a lethal, X-linked muscular disease, due to the loss of dystrophin protein. Antisense oligonucleotide (AON) mediated exon skipping is one of promising approaches to cure this disease and phase II-III clinical trials of exon 51 skipping are ongoing. Preliminary but successful results of these trials are indicating that other exons will be targeted in subsequent trials. To develop effective AON for each DMD case, assays using patient-derived cells are valuable. In some cases, obtaining myoblasts from patients is difficult, therefore utilizing of fibroblasts is considered. We have previously reported the FACS-utilizing MyoD transduction of fibroblasts, and it was practical for exon skipping assay (PLoS One. 2010 Aug 18;5(8):e12239.). Nevertheless, in our current assay system, a long-term differentiation culture is required for enough dystrophin mRNA expression. To address this challenge, we evaluated an availability of several myogenic surface markers. CD56 is known as one of the myogenic markers expressed on a surface of primary myoblasts. We found that the MyoD-transduced fibroblasts, which were initially negative with CD56 by FACS analysis, gradually increased its positive population, and these cells expressed much dystrophin mRNA than negative population. These findings suggest that the expression of myogenic marker protein on MyoD-transduced fibroblasts would be beneficial to efficiently select the cells expressing dystrophin mRNA and shorten the period required for enough dystrophin mRNA expression. We would report the possibility that a stable dystrophin mRNA expression of DMD fibroblasts required for exon skipping assay might be achieved through a combination of MyoD transduction and myogenic surface marker.
Cancer – Immunotherapy I 187. Randomized Phase II Trial of PostOperative Adjuvant Chemotherapy ±FANG™ Autologous Tumor Cell Vaccine in Colorectal Carcinoma with Liver Metastases
Joseph Kuhn,5 Neil Senzer,1,2,3,4 Minal Barve,3 Padmasini Kumar,4 Donald D. Rao,4 Gladice Wallraven,4 Beena O. Pappen,4 Phillip B. Maples,4 John Nemunaitis.1,2,3,4 1 Mary Crowley Cancer Research Centers, Dallas; 2Medical City HCA, Dallas; 3Texas Oncology, P.A., Dallas; 4Gradalis, Inc., Dallas; 5WLS Surgical Associates, P.A., Dallas. The Phase I trial of FANG™ vaccine (comprised of irradiated autologous tumor cells transduced with plasmid encoding GMCSF and the RNA interference bi-shRNA FURIN; see Senzer et al., Mol Ther 2011) involving 31 previously treated solid tumor patients provided sufficient safety data to proceed to Phase II efficacy studies. This decision was bolstered by the correlation of survival with induced immune response (IFNgamma ELISPOT) in serially monitored patients at ≥ 4 months (p=0.025). Specifically, 6 metastatic refractory colon cancer patients in the Phase I trial, who having failed standard therapy, had an expected survival of ∼6 months, received a combined total of 24 vaccinations (i.e. 2 patients at dose level 1 x 107 and 4 patients at 2.5 x 107 cells / injection; data combined as there is no dose related difference in endpoint). These patients currently demonstrate median survivals of 358 days (554, 515+, 413, 303, 185, and 167 days each). Efficacy of the pbi-shRNA™ FURIN vaccine component was confirmed by marked knockdown of the furin proprotein convertase activated immunosuppressive proteins TGFβ1 and b2; 87% and 92% respectively). Based on these results, the Phase II trial (CL-PTL 114, BB-IND 14205) was activated following FDA, RAC, IBC and IRB acceptance. This is a randomized Phase II study of integrated adjuvant modified FOLFOX6 (mFOLFOX6) and intradermal autologous FANG™ cancer vaccine (1.0 x 107 cells/injection; maximum of 12 vaccinations) versus adjuvant mFOLFOX6 and placebo in patients with colorectal carcinoma with either synchronous or metachronous liver metastases (CLM ± pulmonary metastases) following resection ± ablation with curative intent. Eight weeks following resection ± ablation, patients will receive FANG™ Weeks 1, 3 then q 4 weeks with mFOLFOX starting Week 4 then continuing q 2 weeks x 6 months. A minimum harvest aliquot to produce 5 monthly injections will be required for entry into the study. These patients will be managed in an outpatient setting. Hematologic function, liver enzymes, renal function and electrolytes will be monitored weekly. Immune function analyses including assessment of tumor infiltrating lymphocytes (TIL) and ELISPOT analysis of cytotoxic T cell function to autologous tumor antigens will be monitored at baseline (TIL + ELISPOT) and thereafter ELISPOT analysis at Months 2, 4, 6, 9, end of treatment, and at response follow up visits. Preliminary data will be presented at the meeting.
188. Hematopoietic Stem Cell Immunotherapy for Melanoma
Eric H. Gschweng,1 Michael L. Kaufman,1 Roger P. Hollis,1 Koya C. Richard,2 Ribas Antoni,3 Donald B. Kohn.1 1 Molecular Biology, Immunology, and Molecular Genetics, UCLA, Los Angeles, CA; 2Department of Surgery, Surgical Oncology, UCLA, Los Angeles, CA; 3Department of Medicine, Hematology/ Oncology, UCLA, Los Angeles, CA. Melanoma is the deadliest form of skin cancer, and the primary treatment option of surgical removal becomes infeasible with widespread metastasis. Immunotherapy involves redirecting the immune system with transgenic T-cell receptors (TCR), and offers an attractive strategy to combat this disease. While transduction S74
Molecular Therapy Volume 20, Supplement 1, May 2012 Copyright © The American Society of Gene & Cell Therapy