- Email: [email protected]
326. Abstract Withdrawn
Recent Advances and Applications in Adenovirus Vectors and Other DNA Viruses II 3 mRNA was strongly upregulated. In addition, expression of type I, III collagen, fibronectin, and elastin was significantly reduced in dE1-RGD/GFP/DCN-transduced keloid spheroids. Conclusion: These results support the utility of decorin-expressing adenovirus to reduce collagen synthesis in KFs and keloid spheroid, which may be highly beneficial in treating keloids.
325. Persistence of hAQP1 Expression in Human Salivary Gland Cells Following AdhAQP1 Transduction Is Associated With a Lack of Methylation of hCMV Promoter
Changyu Zheng,1 Bruce J. Baum,1 Xibao Liu,1 Corinne M. Goldsmith,1 Paola Perez,1 Shyh-Ing Jang,1 Ana P. Cotrim,1 Linda McCullagh,1 Indu S. Ambudkar,1 Ilias Alevizos.1 1 MPTB/NIDCR, NIH, Bethesda, MD. In 2012, we reported that five out of 11 subjects in a clinical trial (NCT00372320) of AdhAQP1 to radiation-damaged parotid glands showed increased saliva flow rates and decreased symptoms over the initial 42 days. AdhAQP1 is a first generation, E1-deleted, replicationdefective, serotype 5 adenoviral vector encoding human aquaporin-1 (hAQP1). This vector uses the human cytomegalovirus enhancer/ promoter (hCMVp). Since subject responses were longer than expected, we hypothesized that the hCMVp may not be methylated in human salivary gland cells to the extent previously observed in rodent salivary gland cells. To test our hypothesis, initially we transduced mouse and rat cell lines in vitro, or submandibular glands in vivo, with the same AdhAQP1 used in the clinical trial. The hCMVp was gradually methylated over time in rodent cells and associated with a decreased functional hAQP1 expression. The hCMVp, however, was not methylated in human salivary gland primary cultures or human salivary gland cell lines after transduction with AdhAQP1. Importantly, hAQP1 maintained its function in those cells. These data suggest that the hCMVp in AdhAQP1is likely not methylated, resulting in an unexpectedly longer functional expression of hAQP1 in transduced human salivary gland cells.
326.
Abstract Withdrawn
327. Oncolytic Virus Therapy Using Recombinant HSV-1 for Nonseminoma Germ Cell Tumors
Shigenori Kakutani,1 Hiroshi Fukuhara,1 Satoru Taguchi,1 Yuta Takeshima,1 Yukio Homma,1 Yasushi Ino,2 Tomoki Todo.2 1 Department of Urology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan; 2Division of Innovative Cancer Therapy, The Advanced Clinical Reseach Center, The Institution of Medical Science, The University of Tokyo, Tokyo, Japan. Oncolytic herpes simplex viruses type 1 (HSV-1) have been shown promising for treating a variety of tumor. G47∆ is a third-generation oncolytic HSV-1 that has triple mutations in the gamma34.5, alpha47 and ICP6 genes, and the first recombinant HSV-1 tried clinically in Japan. Clinical trials using G47∆ are currently ongoing in patients with glioblastoma, olfactory neuroblastoma or castration resistant prostate cancer. Preclinical studies have shown that G47∆ is also effective in many other types of cancer including breast cancer, schwannoma, renal carcinoma, and urothelial carcinoma. The purpose of this study was to investigate whether oncolytic HSV-1 can also be applied for treating testicular cancer. Testicular cancer is relatively rare, accounting for approximately 1% of male neoplasms and comprising morphologically diverse group of tumors, 90-95% of which are germ cell tumors (GCTs). GCTs are broadly classified as seminoma and non-seminoma (NSGCT). With the increase in early diagnosis and the development of cisplatin-based Molecular Therapy Volume 23, Supplement 1, May 2015 Copyright © The American Society of Gene & Cell Therapy
chemotherapy, a long-term survival can be expected for most GCT patients even with the initial presence of metastases. However, a group of NSGCT patients show resistance to chemotherapy and the five-year survival rate of such group is less than 50%, so clearly a new therapeutic approach is necessary for those patients. In this study, two recombinant HSV-1 were used. T-01 and T-mfIL12 are oncolytic HSV-1 with an empty cassette or the CMV promoterdriven murine interleukin 12 gene inserted into the backbone of G47∆. Human NSGCT cell lines NTERA-2, Tera-1 and ITO-II, and a mouse NSGCT cell line F9 were used for in vitro evaluation. NTERA-2 and F9 were susceptible to T-01 at an MOI of 0.1: The percentages of surviving cells were less than11% at day 3. Cytopathic activities of T-mfIL12 were comparable to T-01. Athymic mice bearing established subcutaneous NTERA-2 tumors and immunocompetent mice bearing established subcutaneous F9 tumors were used for in vivo evaluation. When inoculated intraneoplastically, T-01 (4 x 10^4, 2 x 10^5 or 1 x 10^6 pfu) caused a significant inhibition of tumor growth compared with mock even at the lowest dose in the NTERA-2 model. In the F9 model, T-mfIL12 (4 x 10^4 pfu ) caused a significant inhibition of tumor growth compared with T-01. Moreover, the combination of T-01 (4 x 10^4 pfu) with cisplatin (0.05mg per mouse injected intraperitoneally) was more efficacious than T-01 alone or cisplatin alone in the NTERA-2 model. These data suggest that oncolytic virus therapy using recombinant HSV-1 might be a useful strategy for treating NSGCT.
328. FP3, a Novel VEGF Blocker, Expressing Oncolytic Adenovirus Induces Potent Antiangiogenic and Antitumor Efficacy
Il-Kyu Choi,1 Hyewon Shin,1 Eonju Oh,1 Ji Young Yoo,1 June Kyu Hwang,1 De-Chao Yu,2 Chae-Ok Yun.1 1 Bioengineering, College of Engineering, Hanyang University, Seoul, Korea; 2State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu, China. Various ways to inhibit VEGF, a key facilitator in tumor angiogenesis, are being developed to treat cancer. The soluble VEGF decoy receptor (FP3), due to its high affinity to VEGF, is a highly effective and promising strategy to disrupt VEGF signaling pathway. Despite potential advantage and potent therapeutic efficacy, its employment has been limited by very poor in vivo pharmacokinetic properties. To address this challenge, we designed a novel oncolytic Ad expressing FP3 (RdB/FP3). To demonstrate the VEGF-specific nature of RdB/FP3, replication-incompetent Ad expressing FP3 (dE1/ FP3) was also generated. dE1/FP3 was highly effective in reducing VEGF expression and functionally elicited an antiangiogeneic effect. Furthermore, RdB/FP3 exhibited a potent antitumor effect compared with RdB or recombinant FP3. Consistent with these data, RdB/FP3 was shown to greatly decrease VEGF expression level and vessel density, and increase apoptosis in both tumor endothelial and tumor cells, verifying potent suppressive effects of RdB/FP3 on VEGFmediated tumor angiogenesis in vivo. Importantly, the therapeutic mechanism of antitumor effect mediated by RdB/FP3 is associated with prolonged VEGF silencing efficacy and enhanced oncolysis via cancer cell-specific replication of oncolytic Ad. Taken together, RdB/ FP3 provides a new promising therapeutic approach in the treatment of cancer and angiogenesis-related diseases.
S131
325. Persistence of hAQP1 Expression in Human Salivary Gland Cells Following AdhAQP1 Transduction Is Associated With a Lack of Methylation of hCMV Promoter
Changyu Zheng,1 Bruce J. Baum,1 Xibao Liu,1 Corinne M. Goldsmith,1 Paola Perez,1 Shyh-Ing Jang,1 Ana P. Cotrim,1 Linda McCullagh,1 Indu S. Ambudkar,1 Ilias Alevizos.1 1 MPTB/NIDCR, NIH, Bethesda, MD. In 2012, we reported that five out of 11 subjects in a clinical trial (NCT00372320) of AdhAQP1 to radiation-damaged parotid glands showed increased saliva flow rates and decreased symptoms over the initial 42 days. AdhAQP1 is a first generation, E1-deleted, replicationdefective, serotype 5 adenoviral vector encoding human aquaporin-1 (hAQP1). This vector uses the human cytomegalovirus enhancer/ promoter (hCMVp). Since subject responses were longer than expected, we hypothesized that the hCMVp may not be methylated in human salivary gland cells to the extent previously observed in rodent salivary gland cells. To test our hypothesis, initially we transduced mouse and rat cell lines in vitro, or submandibular glands in vivo, with the same AdhAQP1 used in the clinical trial. The hCMVp was gradually methylated over time in rodent cells and associated with a decreased functional hAQP1 expression. The hCMVp, however, was not methylated in human salivary gland primary cultures or human salivary gland cell lines after transduction with AdhAQP1. Importantly, hAQP1 maintained its function in those cells. These data suggest that the hCMVp in AdhAQP1is likely not methylated, resulting in an unexpectedly longer functional expression of hAQP1 in transduced human salivary gland cells.
326.
Abstract Withdrawn
327. Oncolytic Virus Therapy Using Recombinant HSV-1 for Nonseminoma Germ Cell Tumors
Shigenori Kakutani,1 Hiroshi Fukuhara,1 Satoru Taguchi,1 Yuta Takeshima,1 Yukio Homma,1 Yasushi Ino,2 Tomoki Todo.2 1 Department of Urology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan; 2Division of Innovative Cancer Therapy, The Advanced Clinical Reseach Center, The Institution of Medical Science, The University of Tokyo, Tokyo, Japan. Oncolytic herpes simplex viruses type 1 (HSV-1) have been shown promising for treating a variety of tumor. G47∆ is a third-generation oncolytic HSV-1 that has triple mutations in the gamma34.5, alpha47 and ICP6 genes, and the first recombinant HSV-1 tried clinically in Japan. Clinical trials using G47∆ are currently ongoing in patients with glioblastoma, olfactory neuroblastoma or castration resistant prostate cancer. Preclinical studies have shown that G47∆ is also effective in many other types of cancer including breast cancer, schwannoma, renal carcinoma, and urothelial carcinoma. The purpose of this study was to investigate whether oncolytic HSV-1 can also be applied for treating testicular cancer. Testicular cancer is relatively rare, accounting for approximately 1% of male neoplasms and comprising morphologically diverse group of tumors, 90-95% of which are germ cell tumors (GCTs). GCTs are broadly classified as seminoma and non-seminoma (NSGCT). With the increase in early diagnosis and the development of cisplatin-based Molecular Therapy Volume 23, Supplement 1, May 2015 Copyright © The American Society of Gene & Cell Therapy
chemotherapy, a long-term survival can be expected for most GCT patients even with the initial presence of metastases. However, a group of NSGCT patients show resistance to chemotherapy and the five-year survival rate of such group is less than 50%, so clearly a new therapeutic approach is necessary for those patients. In this study, two recombinant HSV-1 were used. T-01 and T-mfIL12 are oncolytic HSV-1 with an empty cassette or the CMV promoterdriven murine interleukin 12 gene inserted into the backbone of G47∆. Human NSGCT cell lines NTERA-2, Tera-1 and ITO-II, and a mouse NSGCT cell line F9 were used for in vitro evaluation. NTERA-2 and F9 were susceptible to T-01 at an MOI of 0.1: The percentages of surviving cells were less than11% at day 3. Cytopathic activities of T-mfIL12 were comparable to T-01. Athymic mice bearing established subcutaneous NTERA-2 tumors and immunocompetent mice bearing established subcutaneous F9 tumors were used for in vivo evaluation. When inoculated intraneoplastically, T-01 (4 x 10^4, 2 x 10^5 or 1 x 10^6 pfu) caused a significant inhibition of tumor growth compared with mock even at the lowest dose in the NTERA-2 model. In the F9 model, T-mfIL12 (4 x 10^4 pfu ) caused a significant inhibition of tumor growth compared with T-01. Moreover, the combination of T-01 (4 x 10^4 pfu) with cisplatin (0.05mg per mouse injected intraperitoneally) was more efficacious than T-01 alone or cisplatin alone in the NTERA-2 model. These data suggest that oncolytic virus therapy using recombinant HSV-1 might be a useful strategy for treating NSGCT.
328. FP3, a Novel VEGF Blocker, Expressing Oncolytic Adenovirus Induces Potent Antiangiogenic and Antitumor Efficacy
Il-Kyu Choi,1 Hyewon Shin,1 Eonju Oh,1 Ji Young Yoo,1 June Kyu Hwang,1 De-Chao Yu,2 Chae-Ok Yun.1 1 Bioengineering, College of Engineering, Hanyang University, Seoul, Korea; 2State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu, China. Various ways to inhibit VEGF, a key facilitator in tumor angiogenesis, are being developed to treat cancer. The soluble VEGF decoy receptor (FP3), due to its high affinity to VEGF, is a highly effective and promising strategy to disrupt VEGF signaling pathway. Despite potential advantage and potent therapeutic efficacy, its employment has been limited by very poor in vivo pharmacokinetic properties. To address this challenge, we designed a novel oncolytic Ad expressing FP3 (RdB/FP3). To demonstrate the VEGF-specific nature of RdB/FP3, replication-incompetent Ad expressing FP3 (dE1/ FP3) was also generated. dE1/FP3 was highly effective in reducing VEGF expression and functionally elicited an antiangiogeneic effect. Furthermore, RdB/FP3 exhibited a potent antitumor effect compared with RdB or recombinant FP3. Consistent with these data, RdB/FP3 was shown to greatly decrease VEGF expression level and vessel density, and increase apoptosis in both tumor endothelial and tumor cells, verifying potent suppressive effects of RdB/FP3 on VEGFmediated tumor angiogenesis in vivo. Importantly, the therapeutic mechanism of antitumor effect mediated by RdB/FP3 is associated with prolonged VEGF silencing efficacy and enhanced oncolysis via cancer cell-specific replication of oncolytic Ad. Taken together, RdB/ FP3 provides a new promising therapeutic approach in the treatment of cancer and angiogenesis-related diseases.
S131