PURPOSE: This phase I clinical trial evaluated safety, feasibility, and efficiency of nonviral intratumoral jet-injection gene transfer in patients with skin metastases from melanoma and breast cancer. EXPERIMENTAL DESIGN: Seventeen patients were enrolled. The patients received five jet injections with a total dose of 0.05 mg beta-galactosidase (LacZ)-expressing plasmid DNA (pCMVbeta) into a single cutaneous lesion. Clinical and laboratory safety monitoring were done. Systemic plasmid clearance was monitored by quantitative real-time PCR of blood samples throughout the study. All lesions were resected after 2 to 6 days. Intratumoral plasmid DNA load, DNA distribution, and LacZ expression was analyzed by quantitative real-time PCR, quantitative reverse transcription-PCR, Western blot, immunohistochemistry, and 5-bromo-4-chloro-3-indolyl-beta-D-galactoside staining. RESULTS: Jet injection of plasmid DNA was safely done in all patients. No serious side effects were observed. Thirty minutes after jet injection, peak plasmid DNA levels were detected in the blood followed by rapid decline and clearance. Plasmid DNA and LacZ mRNA and protein expression were detected in all treated lesions. Quantitative analysis revealed a correlation of plasmid DNA load and LacZ-mRNA expression confirmed by Western blot. Immunohistochemistry and 5-bromo-4-chloro-3-indolyl-beta-D-galactoside staining showed LacZ-protein throughout the tumor. Transfected tumor areas were found close and distant to the jet-injection site with varying levels of DNA load and transgene expression. CONCLUSION: Intratumoral jet injection of plasmid DNA led to efficient LacZ reporter gene expression in all patients. No side effects were experienced, supporting safety and applicability of this novel nonviral approach. A next step with a therapeutic gene product should determine antitumor efficacy of jet-injection gene transfer.
PURPOSE: This phase I clinical trial evaluated safety, feasibility, and efficiency of nonviral intratumoral jet-injection gene transfer in patients with skin metastases from melanoma and breast cancer. EXPERIMENTAL DESIGN: Seventeen patients were enrolled. The patients received five jet injections with a total dose of 0.05 mg beta-galactosidase (LacZ)-expressing plasmid DNA (pCMVbeta) into a single cutaneous lesion. Clinical and laboratory safety monitoring were done. Systemic plasmid clearance was monitored by quantitative real-time PCR of blood samples throughout the study. All lesions were resected after 2 to 6 days. Intratumoral plasmid DNA load, DNA distribution, and LacZ expression was analyzed by quantitative real-time PCR, quantitative reverse transcription-PCR, Western blot, immunohistochemistry, and 5-bromo-4-chloro-3-indolyl-beta-D-galactoside staining. RESULTS: Jet injection of plasmid DNA was safely done in all patients. No serious side effects were observed. Thirty minutes after jet injection, peak plasmid DNA levels were detected in the blood followed by rapid decline and clearance. Plasmid DNA and LacZ mRNA and protein expression were detected in all treated lesions. Quantitative analysis revealed a correlation of plasmid DNA load and LacZ-mRNA expression confirmed by Western blot. Immunohistochemistry and 5-bromo-4-chloro-3-indolyl-beta-D-galactoside staining showed LacZ-protein throughout the tumor. Transfected tumor areas were found close and distant to the jet-injection site with varying levels of DNA load and transgene expression. CONCLUSION: Intratumoral jet injection of plasmid DNA led to efficient LacZ reporter gene expression in all patients. No side effects were experienced, supporting safety and applicability of this novel nonviral approach. A next step with a therapeutic gene product should determine antitumor efficacy of jet-injection gene transfer.
Authors: Dennis Kobelt; Martin Schleef; Marco Schmeer; Jutta Aumann; Peter M Schlag; Wolfgang Walther Journal: Mol Biotechnol Date: 2013-01 Impact factor: 2.695
Authors: Ronald C Hendrickson; Vito R Cicinnati; Andreas Albers; Grzegorz Dworacki; Andrea Gambotto; Ornella Pagliano; Thomas Tüting; Jose I Mayordomo; Carmen Visus; Ettore Appella; Jeffrey Shabanowitz; Donald F Hunt; Albert B DeLeo Journal: Cancer Immunol Immunother Date: 2009-06-27 Impact factor: 6.968
Authors: Dennis Kobelt; Jutta Aumann; Manuel Schmidt; Burghardt Wittig; Iduna Fichtner; Diana Behrens; Margit Lemm; Greta Freundt; Peter M Schlag; Wolfgang Walther Journal: Mol Oncol Date: 2014-01-18 Impact factor: 6.603
Authors: Tam Nguyen-Hoai; Dennis Kobelt; Oliver Hohn; Minh D Vu; Peter M Schlag; Bernd Dörken; Steven Norley; Martin Lipp; Wolfgang Walther; Antonio Pezzutto; Jörg Westermann Journal: Oncoimmunology Date: 2012-12-01 Impact factor: 8.110