Extracellular and intracellular factors influencing gene transfection mediated by 1,4-dihydropyridine amphiphiles

Hyvönen, Zanna; Ruponen, Marika; Rönkkö, Seppo; Suhonen, Pekka; Urtti, Arto

doi:10.1016/s0928-0987(02)00031-3

Cited by 33 publications

(25 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…More efficient uptake but lower transfection efficiency of PEI-and PLL-polyplexes in comparison to DOTAP-and DOTAP/DOPE lipoplexes has been earlier demonstrated by Ruponen et al [20]. The lack of correlation between the cell uptake and the transgene expression was also demonstrated by others for various polymeric and liposomal carriers [21,22]. This suggests that other intracellular events such as transport to the nucleus and efficient transcription and translation are important in achievement of successful gene expression.…”

Section: Neither Cellular Nor Nuclear Uptake Of Pdna Correlates With mentioning

confidence: 77%

Genetic blockage of endocytic pathways reveals differences in the intracellular processing of non-viral gene delivery systems

Ilina

Hyvönen

Saura

et al. 2012

Journal of Controlled Release

Self Cite

View full text Add to dashboard Cite

Section: Neither Cellular Nor Nuclear Uptake Of Pdna Correlates With mentioning

confidence: 77%

Genetic blockage of endocytic pathways reveals differences in the intracellular processing of non-viral gene delivery systems

Ilina

Hyvönen

Saura

et al. 2012

Journal of Controlled Release

Self Cite

View full text Add to dashboard Cite

“…Positively charged lipoplexes interact with negatively charged cell surface, mediated by proteoglycans, as well as serum or extracellular matrix components, such as soluble glycosaminoglycans. Whereas the former is desirable to achieve cellular uptake, the latter can disrupt the complexes or block transfection [82].…”

Section: (C) Transport Of Lipoplexes To Target Tissuesmentioning

confidence: 99%

“…This both inhibits the lysosomal enzymes that require an optimal acidic pH for activity, but also induces osmotic swelling and burst of the endosome due to water entry [92]. Cationic lipids thought to act (at least partially) by this mechanism include lipopolyamines (Transfectam [92], CTAP [24]), guanidinium cholesterol compound BGTC [23], amidinoamines [93] and 1,4-dihydropyridine-containing lipids [82]. Another strategy uses ingredients that decrease the curvature of the endosomal membrane [94].…”

Section: (E) Endosomal Escapementioning

confidence: 99%

Structure-Activity Relationship in Cationic Lipid Mediated Gene Transfection

Niculescu‐Duvaz¹,

Heyes²,

Springer³

2003

CMC

141

112

View full text Add to dashboard Cite

Non-viral synthetic vectors for gene delivery represent a safer alternative to viral vectors. Their main drawback is the low transfection efficiency, especially in vivo. Among the non-viral vectors currently in use, the cationic liposomes composed of cationic lipids are the most common. This review discusses the physicochemical properties of cationic lipids, the formation, macrostructure and specific parameters of the corresponding formulated liposomes, and the effect of all these parameters on transfection efficiency. The optimisation of liposomal vectors requires both the understanding of the biological variables involved in the transfection process, and the effect of the structural elements of the cationic lipids on these biological variables. The biological barriers relevant for in vitro and in vivo transfection are identified, and solutions to overcome them based on rational design of the cationic lipids are discussed. The review focuses on the relationship between the structure of the cationic lipid and the transfection activity. The structure is analysed in a modular manner. The hydrophobic domain, the cationic head group, the backbone that acts as a scaffold for the other domains, the linkers between backbone, hydrophobic domain and cationic head group, the polyethyleneglycol chains and the targeting moiety are identified as distinct elements of the cationic lipids used in gene therapy. The main chemical functionalities used to built these domains, as well as overall molecular features such as architecture and geometry, are presented. Studies of structure-activity relationships of each cationic lipid domain, including the authors', and the trends identified by these studies, help furthering the understanding of the mechanism governing the formation and behaviour of cationic liposomes in gene delivery, and therefore the rational design of new improved cationic lipids vectors capable of achieving clinical significance.

show abstract

“…This discrepancy in results could not be accounted for by differences in the physical characteristics of A2M and IgG and the other proteins tested, like the molecular weight or pI of the protein. This type of inconsistency between particle uptake and transfection efficiency has been observed in other studies, where exposure of amphiplex vectors to BSA increased vector uptake but inhibited gene expression [27]. One possible explanation for this inconsistency may involve the need for the proper and timely decoupling of the DNA from cationic carrier after cytoplasmic delivery, which may be important for nuclear entry and transcription of the DNA.…”

Section: Discussionmentioning

confidence: 58%

“…Decondensation of DNA may explain the inability of some lipoplexes to transfect cells in the presence of serum, since uncoupling of the DNA from cationic particles may make the DNA more susceptible to enzymatic cleavage and degradation in circulation with systemicallydelivered vectors [12]. Vector decondensation, by individual components such as cell surface proteoglycans (heparan sulfate, chondroitin sulfate and dextran sulfate) [27,28], or in the presence of complete serum [11], has been shown previously, but the specific soluble components within serum that may cause decondensation were not identified. Condensation experiments in this study showed that of all the serum proteins tested, only LDL causes significant release of the DNA from DOTAP/DNA lipoplexes.…”

Section: Discussionmentioning

confidence: 99%

Poly(propylacrylic acid)-mediated serum stabilization of cationic lipoplexes

Cheung¹,

Stayton²,

Hoffman³

2005

Journal of Biomaterials Science, Polymer Edition

View full text Add to dashboard Cite

The serum instability associated with cationic lipoplexes represents one of the major obstacles for the in vivo delivery of nonviral gene therapy vectors. Recently, we have shown that poly(propylacrylic acid) (PPAA), a pH-sensitive polyanionic polymer, can significantly improve the in vitro serum stability of DOTAP lipoplexes and enhance transfection (Cheung et al., Bioconjug. Chem. 12, 906 (2001)). We investigated this serum-stabilizing effect provided by PPAA using methods to identify the specific serum proteins that interact with DOTAP/DNA and DOTAP/DNA/PPAA lipoplexes and determined their modes of interaction with these lipoplexes. Studies showed that only low-density lipoprotein (LDL) caused significant decondensation of DNA from lipoplexes lacking PPAA, but that fully condensed DNA was retained within lipoplexes incorporating PPAA. Another major factor in the loss of transfection activity was due to the reduced cellular uptake of DOTAP lipoplexes upon exposure to serum, with bovine serum albumin (BSA) and high-density lipoprotein (HDL) acting as major contributors to this reduction in vector internalization. In contrast, lipoplexes containing PPAA maintained high levels of uptake into cells in the presence of these proteins. Transfection results generally concurred with the mechanistic studies, suggesting that maintaining effective cellular delivery of intact lipoplexes in the presence of serum proteins is important to retain high transfection efficiencies. These results indicate that the addition of PPAA as a ternary component in DOTAP lipoplexes can overcome some of the serum-related deficiencies encountered with these lipoplexes to provide efficient transfection.

show abstract

Extracellular and intracellular factors influencing gene transfection mediated by 1,4-dihydropyridine amphiphiles

Cited by 33 publications

References 34 publications

Genetic blockage of endocytic pathways reveals differences in the intracellular processing of non-viral gene delivery systems

Genetic blockage of endocytic pathways reveals differences in the intracellular processing of non-viral gene delivery systems

Structure-Activity Relationship in Cationic Lipid Mediated Gene Transfection

Poly(propylacrylic acid)-mediated serum stabilization of cationic lipoplexes

Contact Info

Product

Resources

About