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DNA delivery to 'ex vivo' human liver segments
M J Herrero1, L Sabater, G Guenechea
1Departamento de Farmacología, Facultad de Medicina, Universidad de Valencia, Valencia, Spain.
Gene Therapy
|October 14, 2011
Summary
This study demonstrates effective gene delivery to human liver segments using retrograde hydrodynamic injection. Higher flow rates improved gene expression, suggesting potential for liver gene therapy applications.
Area of Science:
- Hepatology
- Gene Therapy
- Molecular Biology
Background:
- Hydrodynamic injection is effective in rodents but limited in large animals for liver gene therapy.
- An adapted regional hydrodynamic gene delivery system was used to test efficacy in human liver segments.
Purpose of the Study:
- To evaluate the efficacy of hydrodynamic injection for gene delivery in human liver segments.
- To determine optimal flow rates for gene delivery in this model.
Main Methods:
- Human liver segments were obtained via surgical resection and perfused retrogradely from the hepatic vein.
- Saline solution with enhanced green fluorescent protein (eGFP) plasmid was injected at varying flow rates (1, 10, 20 ml/s).
- Gene expression was analyzed via microscopy, fluorescent, immunohistochemistry, and molecular analysis after 1-2 days of culture.
Main Results:
- Gene delivery and expression were observed in liver sections at flow rates ≥10 ml/s.
- Molecular analysis confirmed wide gene expression, with high DNA and RNA copy numbers.
- Gold nanoparticle injection suggested a non-disruptive, facilitated permeation process for DNA delivery to hepatocytes.
Conclusions:
- Retrograde venous injection into watertight human liver segments enables efficient gene delivery and expression.
- Higher flow rates are crucial for achieving widespread gene expression in this model.
- Further research is needed to understand the impact of ischemia on hepatocyte function in this model.
