Expression of recombinant genes in myocardium in vivo after direct injection of DNA.

Abstract

The ability to program recombinant gene expression in cardiac myocytes in vivo holds promise for the treatment of many inherited and acquired cardiovascular diseases. In this report, we demonstrate that a recombinant P-galactosidase gene under the control of the Rous sarcoma virus promoter can be introduced into and expressed in adult rat cardiac myocytes in vivo by the injection of purified plasmid DNA directly into the left ventricular wall. Cardiac myocytes expressing recombinant /3-galactosidase were detected histochemically in rat hearts for at least 4 weeks after injection of the f3-galactosidase gene. These results demonstrate the potential of this method of somatic gene therapy for the treatment of cardiovascular disease. (Circulation 1990;82:2217-2221 Somatic gene therapy, the expression of recombinant genes in non-germ-line tissues of the adult organism, holds great promise for the treatment of many inherited and acquired human diseases (reviewed in Reference 1). The biological requirements for this type of gene therapy include the ability to introduce recombinant genes efficiently into the appropriate cells and tissues and to program the high-level and, in many cases, stable expression of these recombinant genes in vivo. In addition, it is necessary that the process of gene therapy itself not be harmful to the recipient organism, in particular, that the techniques used to introduce the recombinant genes do not result in persistent infection of the host or in deleterious mutations of the recipient cells. Two general approaches have proven useful in animal models of somatic gene therapy. In

Intravascular naked DNA vaccine encoding glycoprotein B induces protective humoral and cellular immunity against herpes simplex virus type 1 infection in mice

“…1,6,7 Virus-specific CTLs have been induced by pDNA vaccine carrying antigens for influenza virus, 19,20 lymphocytic choriomeningitis virus, 21,22 human immunodeficiency virus-1, 23 and hepatitis B 24 and C 25 viruses. When a pDNA vaccine encoding an HSV glycoprotein is immunized into Balb/c 8,[26][27][28][29][30][31][32][33] or Hartley guinea pig 12,34 via the intramuscular route, humoral responses (Th2 and antibody) are predominantly elicited. The CTL response is induced when a Th1 cytokine 35 or the granulocyte-macrophage colony stimulating factor (GM-CSF) 6 is employed as an adjuvant in combination with the pDNA vaccine.…”

“…Live or attenuated HSV vaccine is capable of inducing both antibody and CTL responses. 7 Although humoral responses induced by intramuscular pDNA vaccine protocols protect animals from acute HSV infection, 8,[26][27][28][29][30][31] effectiveness of these strategies in controlling latent and recurrent infection has not been elucidated. The cellular immune responses may play important roles in antivirus defense, in case genetic vaccine will be applied to humans.…”

“…Previous studies on intramuscular genetic vaccination also demonstrated that humoral immune responses mainly contributed to the protection from acute infection. 8,[26][27][28][29][30][31] An intravenous genetic vaccine was tested by Rogers et al 44 who, instead of naked pDNA, combined a gD expression vector with a synthetic macromolecule, asialoorosomucoid poly-L-lysine. The gD antigen was expressed in the parenchymal and nonparenchymal cells of the liver of the injected mice.…”

Sonoporation using microbubble BR14 promotes pDNA/siRNA transduction to murine heart

“…Although non-viral methods are free from virus-associated adverse effects, their transduction efficiency is low. For example, following direct intramyocardial injection of plasmid DNA into the heart of mice [3], rats [4,5], and hamsters [6], the transgenes were expressed only within a small area surrounding the injection site. A more efficient non-viral method of gene transfer is sonoporation of cells.…”

Highly efficient gene transfer into murine liver achieved by intravenous administration of naked Epstein–Barr virus (EBV)-based plasmid vectors

“…The procedure is being tested in some clinical trials, including genetic treatment of cardiovascular diseases. 2 Naked pDNA transfection was also successful into other organs, such as heart, 3 lung, 4 skin, 5 and liver. 6 More recently, very high transgene expression was achieved in the liver by intravascular delivery of pDNA.…”