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504. A Single-Needle Device for Electroporation-Enhanced DNA Delivery
combined with electrochemotherapy induces long-term antitumor immunity. Melanoma Research 2000, 10, 577-583.
The EndoVac
504. A Single-Needle Device for Electroporation-Enhanced DNA Delivery
Feng Lin,' Kate Broderick,' Jonas Soderholm,' Kristen Smits,' Torunn E. Tjelle,' Dietmar Rabussay,' lacob Mathiesen,' Rune
Kjeken .' Research and Development, lnovio Biomedical Corporation, San Diego, CA. I
503. A CE-Marked Automated Device for Electroporation-Enhanced DNA Delivery in Humans Rune Kjeken,' lacob Mathiesen; Feng Lin,' Lei Zhang,' Dietmar Rabussay,' Torunn E. Tjelle. 1 I Research and Development, Inovio Biomedical Corporation, San Diego, CA. In vivo electroporation has emergedas a safe and efficientmethod for enhancing the immunogenictyofplasmid DNA vaccines in large animals and humans. As the technology is now moving into human clinical trials a need for equipment that meets thc requirements for human use has emerged. Such requirements include design and manufacturingto meet regulatorystandards, patient friendlinessand operator ease of use. Cost factors as well as sufficient hardware and software flexibility to allow for implementing future technological and methodological improvementsare also importantconsiderations. Here we present data on ELGEN 1000, a new electroporation system for human use consisting ofa compact square-wave pulse generator and an automated injector.The pulse generator unit has an onboard nash-card-operated PC that interfaces with the user via a touch screen and can record and store treatment protocols and patient data, as well as provide feedback on treatmentresults. The fullyautomated injection/electrode deviceuses off-the-shelfsyringesand needlesand injects the DNA during needle insertion, immediately followed by eleetroporation using the injection needles as electrodes. Injection depth and DNA doses arc programmed via the touch screen. The ELGEN 1000 system is designed and manufactured to meet regulatory requirements for human use and is currently in pre-clinical evaluation for the delivery of pDNA vaccines, including intramuscular deliver)'to non human primates.Cellularand humoral immune responses obtained with the ELGEN 1000 device are comparable with results obtained with multiple needle electrode arrays used in our MedPulser DNA Delivery system. Results obtained with the ELGEN 1000 system will be presented.
SI94
In vivo electroporation (EP) has proven extremely efficient in augmenting the immunogenicity of DNA vaccines in large animal models and has now entered into the first clinical trials for use with therapeutic vaccines in cancer patients. However, current EP proceduresrely on injection needles and multiple needle electrodes, making the delivery device and immunization procedure complex and somewhat painful. For widespread use in prophylactic vaccine delivery, simpler and less invasive procedures may be preferable or required. Here we present data on the design and in vivo evaluation ofa single-needleEPdevice intendedfor the deliveryof prophylactic DNA vaccines. The device is based on a standard 23 G x 38mm hypodermic injection needle. The needle surface was covered with an insulating layer and two eletrodes, separated from each other, were generated by applying thin gold layerson top of the insulating layer. The single-needle device was mounted on a standard I ml syringe and used to inject pDNA intramuscularlyprior to delivery ofelectrical pulses via the same needle.Transgenic expression patterns were first studied using a reporter plasmid encoding Green Fluorescent Protein (GFP). Employing a relatively low current of 100-150 rnA, expression ofGFP in rabbit muscle demonstrated a cylindrical pattern of about 20 mm in length x 5 mm in diameter, surrounding the needle track. The efficiency ofpDNA vaccine delivery was investigated usingpDNAcoding forSecretedAlkalinePhosphatase(SEAP) and human IgG (hlgG). In vivo EP using the single-needle device and 200 ug DNA elicited antibody responses to both antigens, with mean Ab endpoint titers at week 3 reaching> 1Q3 and 1OJ for SEAP and IgG, respectively. In contrast, the same type of immunization without EP failed to elicit significant immune responses toward either of the antigens tested. Our data support the concept that EPenhanced DNA vaccine delivery using a minimalistic single-needle device can generate efficient immunization, potentially providing substantial advantages over presently used devices. NON-VIRAL VECTORS: INTRACELLULAR TRAFFICKING
505. Intracellular Trafficking of Oligonucleotides Delivered with Poly(Glycoamidoamine)s Katye M. Fichter,' Theresa M. Reineke.' I
Department ofChemistry; University ofCincinnati, Cincinnati.
OH.
Efficientintracellulartraffickingof nucleic acids remainsa crucial barrier to the efficacy of nonviral polymeric vectors. As such, it is imperative to understand the impact of chemical structure on the trafficking mechanisms of the delivery vector. An understanding of these mechanisms will contribute greatly to rational designs of future nonviral delivery vectors. This investigation focuses on tracing the intracellular path of a 20bp oligonucleotide decoy delivered with polycations to rat eardiomyoblasts. The polymeric vectors of focus are poly(glycoamidoamine)s (PGAAs), which incorporate sugar moieties into the backbone of a PEl-like repeat unit with 4 secondary amines (Figure I). These vectors diller only in the number of hydroxyl groups and the stereochemistry of their sugar moieties, but have found to have significant differences in transfection efficiency.
Molecular Therapy Yofume 15. Supplement I, .\by 2007 Copyright © '111C American Societyo f Gene Tllcr.lpr
The EndoVac
504. A Single-Needle Device for Electroporation-Enhanced DNA Delivery
Feng Lin,' Kate Broderick,' Jonas Soderholm,' Kristen Smits,' Torunn E. Tjelle,' Dietmar Rabussay,' lacob Mathiesen,' Rune
Kjeken .' Research and Development, lnovio Biomedical Corporation, San Diego, CA. I
503. A CE-Marked Automated Device for Electroporation-Enhanced DNA Delivery in Humans Rune Kjeken,' lacob Mathiesen; Feng Lin,' Lei Zhang,' Dietmar Rabussay,' Torunn E. Tjelle. 1 I Research and Development, Inovio Biomedical Corporation, San Diego, CA. In vivo electroporation has emergedas a safe and efficientmethod for enhancing the immunogenictyofplasmid DNA vaccines in large animals and humans. As the technology is now moving into human clinical trials a need for equipment that meets thc requirements for human use has emerged. Such requirements include design and manufacturingto meet regulatorystandards, patient friendlinessand operator ease of use. Cost factors as well as sufficient hardware and software flexibility to allow for implementing future technological and methodological improvementsare also importantconsiderations. Here we present data on ELGEN 1000, a new electroporation system for human use consisting ofa compact square-wave pulse generator and an automated injector.The pulse generator unit has an onboard nash-card-operated PC that interfaces with the user via a touch screen and can record and store treatment protocols and patient data, as well as provide feedback on treatmentresults. The fullyautomated injection/electrode deviceuses off-the-shelfsyringesand needlesand injects the DNA during needle insertion, immediately followed by eleetroporation using the injection needles as electrodes. Injection depth and DNA doses arc programmed via the touch screen. The ELGEN 1000 system is designed and manufactured to meet regulatory requirements for human use and is currently in pre-clinical evaluation for the delivery of pDNA vaccines, including intramuscular deliver)'to non human primates.Cellularand humoral immune responses obtained with the ELGEN 1000 device are comparable with results obtained with multiple needle electrode arrays used in our MedPulser DNA Delivery system. Results obtained with the ELGEN 1000 system will be presented.
SI94
In vivo electroporation (EP) has proven extremely efficient in augmenting the immunogenicity of DNA vaccines in large animal models and has now entered into the first clinical trials for use with therapeutic vaccines in cancer patients. However, current EP proceduresrely on injection needles and multiple needle electrodes, making the delivery device and immunization procedure complex and somewhat painful. For widespread use in prophylactic vaccine delivery, simpler and less invasive procedures may be preferable or required. Here we present data on the design and in vivo evaluation ofa single-needleEPdevice intendedfor the deliveryof prophylactic DNA vaccines. The device is based on a standard 23 G x 38mm hypodermic injection needle. The needle surface was covered with an insulating layer and two eletrodes, separated from each other, were generated by applying thin gold layerson top of the insulating layer. The single-needle device was mounted on a standard I ml syringe and used to inject pDNA intramuscularlyprior to delivery ofelectrical pulses via the same needle.Transgenic expression patterns were first studied using a reporter plasmid encoding Green Fluorescent Protein (GFP). Employing a relatively low current of 100-150 rnA, expression ofGFP in rabbit muscle demonstrated a cylindrical pattern of about 20 mm in length x 5 mm in diameter, surrounding the needle track. The efficiency ofpDNA vaccine delivery was investigated usingpDNAcoding forSecretedAlkalinePhosphatase(SEAP) and human IgG (hlgG). In vivo EP using the single-needle device and 200 ug DNA elicited antibody responses to both antigens, with mean Ab endpoint titers at week 3 reaching> 1Q3 and 1OJ for SEAP and IgG, respectively. In contrast, the same type of immunization without EP failed to elicit significant immune responses toward either of the antigens tested. Our data support the concept that EPenhanced DNA vaccine delivery using a minimalistic single-needle device can generate efficient immunization, potentially providing substantial advantages over presently used devices. NON-VIRAL VECTORS: INTRACELLULAR TRAFFICKING
505. Intracellular Trafficking of Oligonucleotides Delivered with Poly(Glycoamidoamine)s Katye M. Fichter,' Theresa M. Reineke.' I
Department ofChemistry; University ofCincinnati, Cincinnati.
OH.
Efficientintracellulartraffickingof nucleic acids remainsa crucial barrier to the efficacy of nonviral polymeric vectors. As such, it is imperative to understand the impact of chemical structure on the trafficking mechanisms of the delivery vector. An understanding of these mechanisms will contribute greatly to rational designs of future nonviral delivery vectors. This investigation focuses on tracing the intracellular path of a 20bp oligonucleotide decoy delivered with polycations to rat eardiomyoblasts. The polymeric vectors of focus are poly(glycoamidoamine)s (PGAAs), which incorporate sugar moieties into the backbone of a PEl-like repeat unit with 4 secondary amines (Figure I). These vectors diller only in the number of hydroxyl groups and the stereochemistry of their sugar moieties, but have found to have significant differences in transfection efficiency.
Molecular Therapy Yofume 15. Supplement I, .\by 2007 Copyright © '111C American Societyo f Gene Tllcr.lpr