Intraperitoneal linear polyethylenimine (L-PEI)-mediated gene delivery to ovarian carcinoma nodes in mice

Abstract

Linear polyethylenimine (L-PEI) is an efficient transfection agent for ovarian carcinoma cells in vitro and ex vivo. In the present work, we go a step further and evaluate the efficacy of L-PEI in human ovarian tumor nodes developed in mice. PEI/DNA complexes were administered intraperitoneally instead of intravenously to avoid sequestering of complexes in the lung and liver and to allow transfection of nonvascularized tumor nodes. Plasmid biodistribution was studied by PCR and gene expression was characterized using complementary luciferase and b-galactosidase assays. Intraperitoneal (i.p.) injection of L-PEI/DNA complexes allowed the straightforward distribution of plasmid in the whole peritoneal cavity. Gene expression occurred in many organs, but tumor nodes appeared as preferential sites for transgene expression. The i.p. delivery route allowed repeated injections and administration of large amounts of DNA (up to 400 mg) without signs of toxicity, even for doses well beyond the intravenous lethal dose. Transgene expression was dose-dependent and transient. However, multiple injections allowed its persistence to increase. These results provide encouraging elements towards the development of PEI-based gene therapy protocols for the treatment of advanced stage ovarian carcinoma.

Ultrasound-contrast agent mediated naked gene delivery in the peritoneal cavity of adult rat

“…34,35 Similar to earlier observations that ultrasound increases transfection efficiency in a variety of tissues in vivo including brain, heart and muscle, 12,36,37 we observed increased expression of delivered gene in both parietal and visceral peritoneum following ultrasound exposure. In addition, compared with intraperitoneal gene transfection using polyethylenimine, only very low amount of transgene remained detectable after 72 h, 38 the transgene persists longer in our study signifying the high transfection efficiency by this ultrasound-microbubble-mediated system. We demonstrated for the first time that using ultrasound and microbubble as nonviral transfer agents is safe without eliciting either significant apoptosis by ultrasoundmicrobubble exposure or inflammatory reactions in the peritoneum that are associated with viral vectors.…”

“…Other investigators had observed no toxicity or peritoneal dysfunction with intraperitoneal administration of cationic polymers-DNA, even with amount several folds higher than the intravenous lethal dose. 25,38 Therefore, our findings reveal that the intraperitoneal gene delivery into peritoneal tissues is superior to intravenous route with respect to transfection efficiency and safety.…”

“…It is conceivable that the peritoneal lining not only captures the plasmid DNA but also acts as a barrier preventing the spread of transgene into the underlying organ parenchyma. 38,46 Hence, the ratio of untransfected cells in these organs to the transfected cells in the peritoneal lining may be too large to detect the effective transfection in these organs. A successful transfection method must be a means by which the expression of the transgene can be controlled.…”

Systemic in vivo delivery of siRNA to tumours using combination of polyethyleneimine and transferrin–polyethyleneimine conjugates

“…40,45 In a study using PEIcomplexed siRNA administered intraperitoneally and a subcutaneous ovarian cancer xenograft model, a HER-2 specific siRNA significantly reduced tumour growth over 14 days compared with a control siRNA. 40 Further studies would be needed to verify the low efficiency of delivery by the intravenous route observed in our small pilot study, to determine the underlying mechanisms and to investigate whether intravenous delivery of TfPEI-siRNA complexes can be achieved; however, the current study provides the proof-of-principle that systemic delivery using TfPEI-siRNA complexes is feasible.…”

Cyclodextrin-scaffolded amphiphilic aminoglucoside clusters: self-assembling and gene delivery capabilities

“…The quantities of compound used were calculated according to the desired pDNA concentration of 0.1 mg mL À1 (303 mM phosphate), the N/P ratio, the molar weight and the number of protonable nitrogens in the selected CD derivative or Jet-PEI. 47,48 Experiments were performed for N/P 5 and 10. Concerning the preparation of the DNA complexes from CD derivatives and Jet-PEI, pDNA was diluted in HEPES (20 mM, pH 7.4) to a final concentration of 303 mM, and then the desired amount of CD derivative was added from 10 or 20 mM stock solution (DMSO).…”