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645. Bioreducible Polymer-Conjugated Oncolytic Adenovirus for Hepatoma-Specific Therapy Via Systemic Administration
CANCER - ONCOLYTIC VIRUSES II 644. Therapeutic Evaluation of Prostate Cancer Specific Oncolytic Adenovirus Armed with Fusion Protein Gene PSA-IZ-CD40L
Yue-feng Yang,1 Shu-Ya Xue,1 Hua Wang,1 Feng-jun Xiao,1 Yue Yin,1 Zhuo-zhuang Lu,1 Qun-wei Zhang,1 Chu-tse Wu,1 Li-sheng Wang.1 1 Beijing Institute of Radiation Medicine, Beijing, China.
Prostate cancer is the second most common cancer in men worldwide. Metastatic, castration-resistant prostate cancer remains incurable and novel strategies are urgently needed. Immunotherapy and gene therapy has emerged as promising approaches for treatment of advanced prostate cancers. Oncolytic adenovirus can selectively replicate in and lead to tumor cell lysis, and carry the immune gene to evoke the immune response against cancer. In this study, we constructed and developed a prostate cancer specific oncolytic adenovirus carrying immune gene activating CD40 signals and evaluated its therapeutic effect in vitro and in vivo. Firstly, we constructed two prostate cancer specific oncolytic adenovirus in which prostate cancer specific PPT promoter drives expression of E1A (Ad-PPT-E1A) and armed with a fusion gene of PSA and CD40 Ligand (PL) (Ad-PL-PPT-E1A). Both Ad-PL-PPT-E1A and Ad-PPT-E1A could selectively kill prostate cancer cells effectively in a dose-dependent manner. Treatment of DU145 and PC3M prostate cell line with Ad-PL-PPT-E1A at 10 MOI resulted in 75.23±4.93% and 82.69±1.35% cell death respectively. Whereas this virus shows little effect on none prostate cells (EC304 cells 3.20±0.21% and L02 cells 3.36±6.44 % cell death). Ad-PL-PPT-E1A or Ad-PL-E1A also induce apoptosis in DU145 and PC3M cells obviously in a dose- and time-dependent manner. After infected by Ad-PL-PPT-E1A at 50MOI, the apoptosis rate of PC3M cells rose from 11.02±0.63% (24hours) to 88.99±1.16% (72hours). We also confirmed the prostate cancer specific replication of these two virus by E1A expression using realtime PCR and progeny viruses assay. In general, they could mediate the replication of viruses in prostate cancer cells, but not in other cells. In prostate cancer cells, the increase of viral titers correlated with culture period, and the titer reached nearly 100 IU per cell at 72h post-infection. CD40-CD40 ligand interactions are important for various dendritic cell (DC) functions. Peripheral blood derived iDCs were induced by LPS or cell lysate of adenovirus infected prostate cancer cells. The expression of CD80, CD83 and CD86 in Ad-PL group or Ad-PL-PPT-E1A group was increased obviously similar to LPS-induced positive control. Finally, we evaluated the therapeutic effects of Ad-PL-PPT-E1A on prostate cancer by using PC3 cell exnograft tumor models in nude mice. Ad-PL-PPT-E1A could efficiently replicate in tumor, and the median infectious titers in tumor could reach 1.18×106(4.63×104-2.5×106) IU/tumor 4 days after injected intratumoral, while that was 1.75×103(6.25×102-4.0×103) IU/ tumor in Ad-Null group. And Ad-PL-PPT-E1A inhibited tumor growth obviously. At day 20 post-treatment, the volume of tumor in Ad-Null group increased nearly 1.89±0.58 times while that of Ad-PL-PPTE1A group increased 0.41±0.83 times. Treatment of Ad-PL-PPT-E1A also significantly reduced liver metastasis of PC3 cells.In conclusion, we have successfully developed a prostate cancer specific oncolytic adenovirus Ad-PL-PPT-E1A which could mediate oncolytic effect and activate DCs. These results indicated that it might be an alternative approach for prostate cancer.
S246
645. Bioreducible Polymer-Conjugated Oncolytic Adenovirus for Hepatoma-Specific Therapy Via Systemic Administration
Pyung-Hwan Kim,1,2 Jaesung Kim,1 Tae-il Kim,2 Hye Yeong Nam,1 James W. Yockman,1 Minjung Kim,3 Sung Wan Kim,1,4 Chae-Ok Yun.4 1 Department of Pharmaceutics and Pharmaceutical Chemistry, University of Utah, Salt Lake City; 2Department of Biosystems and Biomaterials Science and Engineering, Seoul National University, Seoul, Republic of Korea; 3Brain Korea 21 Project for Medical Sciences, Yonsei University College of Medicine, Seoul, Republic of Korea; 4Department of Bioengineering, Hanyang University, Seoul, Republic of Korea. Systemic administration of adenovirus (Ad) vectors is complicated by host immune responses and viral accumulation in the liver, resulting in a short circulatory virus half-life, low efficacy, and host side effects. Ad surface modification is thus required to enhance safety and therapeutic efficacy. An arginine-grafted bioreducible polymer (ABP) was chemically conjugated to the Ad surface, generating Ad-E1/GFP-ABP. A hepatocellular carcinoma [HCC]selective oncolytic Ad complex, YKL-1001-ABP, was also generated. Transduction efficiency of Ad-E1/GFP-ABP was enhanced compared to naked Ad-E1/GFP. YKL-1001-ABP elicited an enhanced and specific killing effect in liver cancer cells (Huh7 and HepG2) expressing -fetoprotein (AFP). Compared with naked Ad, systemic administration of ABP-conjugated Ad resulted in reduced liver toxicity and interleukin (IL)-6 production in vitro and in vivo. Ad-E1/GFP-ABP was more resistant to the neutralizing effects of human serum compared to naked Ad-E1/GFP. ABP conjugation extended blood circulation time 45-fold and reduced anti-Ad Ab neutralization. Moreover, systemic administration of YKL-1001ABP markedly suppressed growth of Huh7 hepatocellular carcinoma. These results demonstrate that chemical conjugation of ABP to the Ad surface improves safety and efficacy, indicating that ABP-conjugated Ad is a potentially useful cancer therapeutic agent to target cancer via systemic administration.
646. Human Bone Marrow-Derived Mesenchymal Stem Cells for Intranasal Delivery of Oncolytic Herpes Simplex Virus Type I to Human Gliomas
Tyrel T. Smith,1 Justin C. Roth,1 James M. Markert,1 Richard J. Whitley.1 1 University of Alabama at Birmingham, Birmingham, AL.
Gliomas are the most common and fatal malignancy of the adult central nervous system. The need for new therapeutic options is clear, as standard of care therapies only extend median survival 1214 months beyond diagnosis. Conditionally replication-competent oncolytic herpes simplex type 1 viruses (oHSV) have emerged as promising therapeutics for treating malignant gliomas. The neurotropic properties of HSV-1 make it ideal for targeting brain tumors, and mutations in the diploid 134.5 neurovirulence genes of HSV-1 restrict viral replication to tumor cells. Viral replication in the malignant cells results in oncolysis and release of mutated oHSV progeny, which can then infect neighboring tumor cells and amplify the therapeutic benefit. Furthermore, independent Phase I and Phase Ib trials have reported safe administration of oHSV to patients with malignant glioma. However, efficient delivery and tumor targeting remain as key barriers to therapeutic efficacy. Administration of oHSV is typically limited to the tumor resection cavity immediately following surgery, and virus injected intravenously is rapidly degraded by complement and antibodies and is impervious to the blood brain barrier. Mesenchymal stem cells (MSCs) have been identified as potential vehicles for delivering therapeutics to a Molecular Therapy Volume 21, Supplement 1, May 2013 Copyright © The American Society of Gene & Cell Therapy
Yue-feng Yang,1 Shu-Ya Xue,1 Hua Wang,1 Feng-jun Xiao,1 Yue Yin,1 Zhuo-zhuang Lu,1 Qun-wei Zhang,1 Chu-tse Wu,1 Li-sheng Wang.1 1 Beijing Institute of Radiation Medicine, Beijing, China.
Prostate cancer is the second most common cancer in men worldwide. Metastatic, castration-resistant prostate cancer remains incurable and novel strategies are urgently needed. Immunotherapy and gene therapy has emerged as promising approaches for treatment of advanced prostate cancers. Oncolytic adenovirus can selectively replicate in and lead to tumor cell lysis, and carry the immune gene to evoke the immune response against cancer. In this study, we constructed and developed a prostate cancer specific oncolytic adenovirus carrying immune gene activating CD40 signals and evaluated its therapeutic effect in vitro and in vivo. Firstly, we constructed two prostate cancer specific oncolytic adenovirus in which prostate cancer specific PPT promoter drives expression of E1A (Ad-PPT-E1A) and armed with a fusion gene of PSA and CD40 Ligand (PL) (Ad-PL-PPT-E1A). Both Ad-PL-PPT-E1A and Ad-PPT-E1A could selectively kill prostate cancer cells effectively in a dose-dependent manner. Treatment of DU145 and PC3M prostate cell line with Ad-PL-PPT-E1A at 10 MOI resulted in 75.23±4.93% and 82.69±1.35% cell death respectively. Whereas this virus shows little effect on none prostate cells (EC304 cells 3.20±0.21% and L02 cells 3.36±6.44 % cell death). Ad-PL-PPT-E1A or Ad-PL-E1A also induce apoptosis in DU145 and PC3M cells obviously in a dose- and time-dependent manner. After infected by Ad-PL-PPT-E1A at 50MOI, the apoptosis rate of PC3M cells rose from 11.02±0.63% (24hours) to 88.99±1.16% (72hours). We also confirmed the prostate cancer specific replication of these two virus by E1A expression using realtime PCR and progeny viruses assay. In general, they could mediate the replication of viruses in prostate cancer cells, but not in other cells. In prostate cancer cells, the increase of viral titers correlated with culture period, and the titer reached nearly 100 IU per cell at 72h post-infection. CD40-CD40 ligand interactions are important for various dendritic cell (DC) functions. Peripheral blood derived iDCs were induced by LPS or cell lysate of adenovirus infected prostate cancer cells. The expression of CD80, CD83 and CD86 in Ad-PL group or Ad-PL-PPT-E1A group was increased obviously similar to LPS-induced positive control. Finally, we evaluated the therapeutic effects of Ad-PL-PPT-E1A on prostate cancer by using PC3 cell exnograft tumor models in nude mice. Ad-PL-PPT-E1A could efficiently replicate in tumor, and the median infectious titers in tumor could reach 1.18×106(4.63×104-2.5×106) IU/tumor 4 days after injected intratumoral, while that was 1.75×103(6.25×102-4.0×103) IU/ tumor in Ad-Null group. And Ad-PL-PPT-E1A inhibited tumor growth obviously. At day 20 post-treatment, the volume of tumor in Ad-Null group increased nearly 1.89±0.58 times while that of Ad-PL-PPTE1A group increased 0.41±0.83 times. Treatment of Ad-PL-PPT-E1A also significantly reduced liver metastasis of PC3 cells.In conclusion, we have successfully developed a prostate cancer specific oncolytic adenovirus Ad-PL-PPT-E1A which could mediate oncolytic effect and activate DCs. These results indicated that it might be an alternative approach for prostate cancer.
S246
645. Bioreducible Polymer-Conjugated Oncolytic Adenovirus for Hepatoma-Specific Therapy Via Systemic Administration
Pyung-Hwan Kim,1,2 Jaesung Kim,1 Tae-il Kim,2 Hye Yeong Nam,1 James W. Yockman,1 Minjung Kim,3 Sung Wan Kim,1,4 Chae-Ok Yun.4 1 Department of Pharmaceutics and Pharmaceutical Chemistry, University of Utah, Salt Lake City; 2Department of Biosystems and Biomaterials Science and Engineering, Seoul National University, Seoul, Republic of Korea; 3Brain Korea 21 Project for Medical Sciences, Yonsei University College of Medicine, Seoul, Republic of Korea; 4Department of Bioengineering, Hanyang University, Seoul, Republic of Korea. Systemic administration of adenovirus (Ad) vectors is complicated by host immune responses and viral accumulation in the liver, resulting in a short circulatory virus half-life, low efficacy, and host side effects. Ad surface modification is thus required to enhance safety and therapeutic efficacy. An arginine-grafted bioreducible polymer (ABP) was chemically conjugated to the Ad surface, generating Ad-E1/GFP-ABP. A hepatocellular carcinoma [HCC]selective oncolytic Ad complex, YKL-1001-ABP, was also generated. Transduction efficiency of Ad-E1/GFP-ABP was enhanced compared to naked Ad-E1/GFP. YKL-1001-ABP elicited an enhanced and specific killing effect in liver cancer cells (Huh7 and HepG2) expressing -fetoprotein (AFP). Compared with naked Ad, systemic administration of ABP-conjugated Ad resulted in reduced liver toxicity and interleukin (IL)-6 production in vitro and in vivo. Ad-E1/GFP-ABP was more resistant to the neutralizing effects of human serum compared to naked Ad-E1/GFP. ABP conjugation extended blood circulation time 45-fold and reduced anti-Ad Ab neutralization. Moreover, systemic administration of YKL-1001ABP markedly suppressed growth of Huh7 hepatocellular carcinoma. These results demonstrate that chemical conjugation of ABP to the Ad surface improves safety and efficacy, indicating that ABP-conjugated Ad is a potentially useful cancer therapeutic agent to target cancer via systemic administration.
646. Human Bone Marrow-Derived Mesenchymal Stem Cells for Intranasal Delivery of Oncolytic Herpes Simplex Virus Type I to Human Gliomas
Tyrel T. Smith,1 Justin C. Roth,1 James M. Markert,1 Richard J. Whitley.1 1 University of Alabama at Birmingham, Birmingham, AL.
Gliomas are the most common and fatal malignancy of the adult central nervous system. The need for new therapeutic options is clear, as standard of care therapies only extend median survival 1214 months beyond diagnosis. Conditionally replication-competent oncolytic herpes simplex type 1 viruses (oHSV) have emerged as promising therapeutics for treating malignant gliomas. The neurotropic properties of HSV-1 make it ideal for targeting brain tumors, and mutations in the diploid 134.5 neurovirulence genes of HSV-1 restrict viral replication to tumor cells. Viral replication in the malignant cells results in oncolysis and release of mutated oHSV progeny, which can then infect neighboring tumor cells and amplify the therapeutic benefit. Furthermore, independent Phase I and Phase Ib trials have reported safe administration of oHSV to patients with malignant glioma. However, efficient delivery and tumor targeting remain as key barriers to therapeutic efficacy. Administration of oHSV is typically limited to the tumor resection cavity immediately following surgery, and virus injected intravenously is rapidly degraded by complement and antibodies and is impervious to the blood brain barrier. Mesenchymal stem cells (MSCs) have been identified as potential vehicles for delivering therapeutics to a Molecular Therapy Volume 21, Supplement 1, May 2013 Copyright © The American Society of Gene & Cell Therapy