Image-guided, Intravascular Hydrodynamic Gene Delivery to Skeletal Muscle in Pigs

Department of Pharmaceutical Sciences, School of Pharmacy, University of Pittsburgh, Pittsburgh, Pennsylvania 15261, USA.
Molecular Therapy (Impact Factor: 6.23). 10/2009; 18(1):93-100. DOI: 10.1038/mt.2009.206
Source: PubMed


Development of an effective, safe, and convenient method for gene delivery to muscle is a critical step toward gene therapy for muscle-associated diseases. Toward this end, we have explored the possibility of combining the image-guided catheter insertion technique with the principle of hydrodynamic delivery to achieve muscle-specific gene transfer in pigs. We demonstrate that gene transfer efficiency of the procedure is directly related to flow rate, injection pressure, and injection volume. The optimal gene delivery was achieved at a flow rate of 15 ml/second with injection pressure of 300 psi and injection volume equal to 1.5% of body weight. Under such a condition, hydrodynamic injection of saline containing pCMV-Luc (100 microg/ml) resulted in luciferase activity of 10(6) to 10(7) relative light units (RLU)/mg of proteins extracted from the targeted muscle 5 days after hydrodynamic gene delivery. Result from immunohistochemical analysis revealed 70-90% transfection efficiency of muscle groups in the hindlimb and persistent reporter gene expression for 2 months in transfected cells. With an exception of transient edema and elevation of creatine phosphokinase, no permanent tissue damage was observed. These results suggest that the image-guided, intravenous hydrodynamic delivery is an effective and safe method for gene delivery to skeletal muscle.

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Available from: Kenya Kamimura, Feb 11, 2014
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    • "These results suggest that catheter-based regional hydrodynamic gene delivery has a great potential in treating diseases that cannot be treated with conventional therapy. Since early studies focused on the effectiveness of the procedure [8], [9], [11], a thorough analysis of the safety of the procedure is needed. Therefore, in this study, we aim to perform a more thorough assessment of the safety from a clinical point of view. "
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    ABSTRACT: Evidence in support of safety of a gene delivery procedure is essential toward gene therapy. Previous studies using the hydrodynamics-based procedure primarily focus on gene delivery efficiency or gene function analysis in mice. The current study focuses on an assessment of the safety of computer-controlled and liver-targeted hydrodynamic gene delivery in dogs as the first step toward hydrodynamic gene therapy in clinic. We demonstrate that the impacts of the hydrodynamic procedure were limited in the injected region and the influences were transient. Histological examination and the hepatic microcirculation measurement using reflectance spectrophotometry reveal that the liver-specific impact of the procedure involves a transient expansion of the liver sinusoids. No systemic damage or toxicity was observed. Physiological parameters, including electrocardiogram, heart rate, blood pressure, oxygen saturation, and body temperature, remained in normal ranges during and after hydrodynamic injection. Body weight was also examined to assess the long-term effects of the procedure in animals who underwent 3 hydrodynamic injections in 6 weeks with 2-week time interval in between. Serum biochemistry analysis showed a transient increase in liver enzymes and a few cytokines upon injection. These results demonstrate that image-guided, liver-specific hydrodynamic gene delivery is safe.
    Full-text · Article · Oct 2014 · PLoS ONE
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    • "The biggest challenge over the last few years has been in the identification of the factors and parameters that have important roles in determining the safety and efficiency of hydrodynamic gene delivery to the liver. Previously, we and others have reported that hydrodynamic gene delivery can be applied for gene delivery to the liver, muscle, and kidney in small pigs and rabbits.7,8,9,10,11,12,18 Of note, Herrero et al. have demonstrated in a more recent study that effective hydrodynamic gene delivery is also achievable ex vivo in the human liver.13 "
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    ABSTRACT: Development of a safe and effective method for gene delivery to hepatocytes is a critical step toward gene therapy for liver diseases. Here, we assessed the parameters for gene delivery to the livers of large animals (pigs, 40-65 kg) using an image-guided hydrodynamics-based procedure that involves image-guided catheter insertion into the lobular hepatic vein and hydrodynamic injection of reporter plasmids using a computer-controlled injector. We demonstrated that injection parameters (relative position of the catheter in the hepatic vasculature, intravascular pressure upon injection, and injection volume) are directly related to the safety and efficiency of the procedure. By optimizing these parameters, we explored for the first time, the advantage of the procedure for sequential injections to multiple lobes in human-sized pigs. The optimized procedure resulted in sustained expression of the human α-1 antitrypsin gene in livers for more than 2 months after gene delivery. In addition, repeated hydrodynamic gene delivery was safely conducted and no adverse events were seen in the entire period of the study. Our results support the clinical applicability of the image-guided hydrodynamic gene delivery method for the treatment of liver diseases.Molecular Therapy-Nucleic Acids (2013) 2, e128; doi:10.1038/mtna.2013.52; published online 15 October 2013.
    Full-text · Article · Oct 2013 · Molecular Therapy - Nucleic Acids
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    • "Hydrodynamic gene delivery is an effective method of gene delivery20,21 and has been widely used for gene expression and functional analysis in whole animals.22 The use of the computer-controlled injection device in recent work has made the hydrodynamics-based procedure more attainable for gene delivery in both small and large animals.23–25 Results in Figure 1 show that hydrodynamic injection of pCMV-Acrp30 plasmid results in a significant production of adiponectin as early as 2 h. "
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    ABSTRACT: Adiponectin and its receptors are inversely related to the degree of obesity and have been identified as potential therapeutic targets for the treatment of obesity. In this study, we evaluated the effect of hydrodynamic delivery of adiponectin and/or its receptor 2 (adipoR2) genes on controlling the development of obesity and insulin resistance in AKR/J mice fed a high-fat diet. An increase in adiponectin and adipoR2 gene expression by hydrodynamic gene delivery prevented diet-induced weight gain, reduced fat accumulation in liver and adipose tissue, and improved insulin sensitivity. Beneficial effects were seen with reduced gluconeogenesis in the liver and lipogenesis in the liver, white adipose tissue and skeletal muscle. Real-time PCR analysis demonstrated overexpression of adiponectin and adipoR2 significantly suppressed transcription of phosphoenolpyruvate carboxykinase (pepck), glucose-6-phosphatase (g6pase), stearoyl CoA desaturase 1 (scd-1) and fatty acid synthase (fas) gene. Inhibition effects were mediated by activating the AMP-activated protein kinase (AMPK). These results prove that elevation of adiponectin and/or adipoR2 expression via gene transfer is an effective approach in managing obesity epidemics.Gene Therapy advance online publication, 21 February 2013; doi:10.1038/gt.2013.8.
    Full-text · Article · Feb 2013 · Gene therapy
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