Increased gene expression after liposome-mediated arterial gene transfer associated with intimal smooth muscle cell proliferation. In vitro and in vivo findings in a rabbit model of vascular injury.

Abstract

Arterial gene transfer represents a novel strategy that is potentially applicable to a variety of cardiovascular disorders. Attempts to perform arterial gene transfer using nonviral vectors have been compromised by a low transfection efficiency. We investigated the hypothesis that cellular proliferation induced by arterial injury could augment gene expression after liposome-mediated gene transfer. Nondenuded and denuded rabbit arterial strips were maintained in culture for up to 21 d, after which transfection was performed with a mixture of the plasmid encoding firefly luciferase and cationic liposomes. In nondenuded arteries, the culture interval before transfection did not affect the gene expression. In contrast, denuded arteries cultured for 3-14 d before transfection yielded 7-13-fold higher expression (vs. day 0; P < 0.005). Transfection was then performed percutaneously to the iliac arteries of live rabbits with or without antecedent angioplasty. Gene expression increased when transfection was performed 3-.7 d postangioplasty (P < 0.05). Proliferative activity of neointimal cells assessed in vitro by 13Hlthymidine incorporation, and in vivo by immunostaining for proliferating cell nuclear antigen, increased and declined in parallel with gene expression. These findings thus indicate that the expression of liposome-mediated arterial gene transfer may be augmented in presence of ongoing cellular proliferation. (J. Clin. Invest. 1994. 93:652461.)

Improvement of exogenous DNA nuclear importation by nuclear localization signal‐bearing vectors: a promising way for non‐viral gene therapy?

“…Other results indicate that mitotic activity enhances transfection not only by lipoplexes but also by polyplexes, but not a viral system, which has an efficient nuclear entry machinery, suggesting that transfection close to M phase is facilitated by nuclear membrane breakdown Brunner et al, 2000;Escriou et al, 2001;Ludtke et al, 2002). Such a correlation between transfection efficiency and mitosis has also been reported for in vivo experiments (Takeshita et al, 1994;Vitadello et al, 1994). In contrast to these observations, Brunner et al (2002) have observed that unlike other particle-mediated delivery methods, the linear polyethylenimine (PEI) led to only small differences in gene transfer between HeLa cells transfected in G1 and those in S/G2.…”

Polyethylenimine-mediated gene transfer into pancreatic tumor dissemination in the murine peritoneal cavity

“…An additional element which may account for the apparent tumor-preference is that the transduced gene is more readily expressed in rapidly proliferating cells such as cancer cells than in normal cells with low mitogenic activity. 30,34,35 Since transfected DNA is probably transported inefficiently through the nuclear membrane during interphase, it is likely that the breakdown of the nuclear membrane during mitosis is crucial to efficient entrance of the transfected DNA into the nucleus. 36,37 Issues pertaining to the toxicity, kinetics and biodistribution of DNA:PEI complexes need to be addressed.…”

Efficient lipid-mediated gene transfer to articular chondrocytes

“…Studies in other cell types have demonstrated that optimal lipofection requires mitotic activity. 33,34 A variety of other factors, including differences in cell-specific endosomal and lysosomal activity, nuclear transport and nuclear stability of exogenous DNA may also contribute to these observed differences. 35 Transfection efficiency may also be influenced by duration of exposure and it is possible that further optimization may be achievable by modulating lipid/DNA exposure time to the cells.…”