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Novel jet-injection technology for nonviral intratumoral gene transfer in patients with melanoma and breast cancer

Item Type:Article
Title:Novel jet-injection technology for nonviral intratumoral gene transfer in patients with melanoma and breast cancer
Creators Name:Walther, W. and Siegel, R. and Kobelt, D. and Knoesel, T. and Dietel, M. and Bembenek, A. and Aumann, J. and Schleef, M. and Baier, R. and Stein, U. and Schlag, P.M.
Abstract: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.
Keywords:Western Blotting, Breast Neoplasms, Gene Therapy, Immunohistochemistry, Jet Injections, Lac Operon, Melanoma, Plasmids, Messenger RNA, Reverse Transcriptase Polymerase Chain Reaction, Skin Neoplasms
Source:Clinical Cancer Research
ISSN:1078-0432
Publisher:American Association for Cancer Research (U.S.A.)
Volume:14
Number:22
Page Range:7545-7553
Date:15 November 2008
Official Publication:https://doi.org/10.1158/1078-0432.CCR-08-0412
PubMed:View item in PubMed

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