Mechanism of liver gene transfer by mechanical massage

Many metabolic diseases are caused by defects in the metabolic pathways in the liver. Others result from the absence of specific proteins normally produced and secreted by the liver. Because these metabolic disorders are usually caused by single gene defect, they are ideal candidates for gene therapy. We have previously shown that mouse liver can be transfected by mechanically massaging the liver (MML) after intravenous injection of naked plasmid DNA. We now show a significant linear relationship between the level of liver gene expression and the venous blood pressure, supporting the idea that gene transfer by MML is due, at least in part, to pressure-mediated effect. Liver transfection could not be blocked by co-injection of excess irrelevant DNA or poly I, suggesting that there is no involvement of receptors, including the scavenger receptor, in MML. Moreover, the level of gene expression could be further enhanced by a combination of MML and an increase in DNA retention-time in the liver. Persistence of gene expression could be significantly improved using an EBV-based plasmid vector. Our data suggest the mechanical massage produces transient membrane defects through which naked DNA can enter into the liver cells by simple diffusion.