Enhanced gene expression of systemically administered plasmid DNA in the liver with therapeutic ultrasound and microbubbles
| Autor: | Balasundara Raju, Ralf Seip, Xiaoyan Luo, Songtao Li, Dwight D. Koeberl, Shriram Sethuraman, Andrew Bird, Evgeniy Leyvi |
|---|---|
| Rok vydání: | 2013 |
| Předmět: |
Acoustics and Ultrasonics
Genetic enhancement medicine.medical_treatment Blotting Western Pilot Projects Pharmacology Transfection Mice Western blot Gene expression medicine Animals Ultrasonics Electrical and Electronic Engineering Instrumentation Mice Knockout Microbubbles medicine.diagnostic_test Therapeutic ultrasound business.industry DNA Genetic Therapy Reverse transcription polymerase chain reaction Liver Knockout mouse Glucose-6-Phosphatase Systemic administration business Plasmids |
| Zdroj: | IEEE Transactions on Ultrasonics, Ferroelectrics and Frequency Control. 60:88-96 |
| ISSN: | 0885-3010 |
| DOI: | 10.1109/tuffc.2013.2540 |
| Popis: | Ultrasound-mediated delivery (USMD) of novel therapeutic agents in the presence of microbubbles is a potentially safe and effective method for gene therapy offering many desired characteristics, such as low toxicity, potential for repeated treatment, and organ specificity. In this study, we tested the capability of USMD to improve gene expression in mice livers using glycogen storage disease Type Ia as a model disease under systemic administration of naked plasmid DNA. Image-guided therapeutic ultrasound was used in two studies to provide therapeutic ultrasound to mice livers. In the first study, involving wild-type mice, control animals received naked plasmid DNA (pG6Pase 150 μg) via the tail vein, followed by an infusion of microbubbles; the treated animals additionally received therapeutic ultrasound (1 MHz). Following the procedure, the animals were left to recover and were subsequently euthanized after 2 d and liver samples were extracted. Reverse transcription polymerase chain reaction (RT-PCR) assays were performed on the samples to quantify mRNA expression. In addition, Western blot assays of FLAG-tagged glucose-6-phosphatase (G6Pase) were performed to evaluate protein expression. Ultrasound-exposed animals showed a 4-fold increase in G6Pase RNA in the liver, in comparison with control animals. Furthermore, results from Western blot analysis demonstrated a 2-fold increased protein expression in ultrasound-exposed animals after two days ( p |
| Databáze: | OpenAIRE |
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