Cell-surface glycosaminoglycans inhibit intranuclear uptake but promote post-nuclear processes of polyamidoamine dendrimer–pDNA transfection

Ziraksaz, Zarrintaj; Nomani, Alireza; Ruponen, Marika; Soleimani, Masoud; Tabbakhian, Majid; Haririan, Ismaeil

doi:10.1016/j.ejps.2012.10.016

Cited by 16 publications

(15 citation statements)

References 44 publications

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“…15–17 In order to determine if differences in polyplex morphology affect electrostatic interactions with HSPGs, cellular uptake of polyplexes formulated with [ 3 H]DNA was assessed in wild-type Chinese hamster ovary (CHO) cell lines normally expressing HSPGs on their extracellular surface (CHO-K1) or mutant CHO cells lacking the presence of HSPGs (CHO-pgs-A745). 16 Surprisingly, polyplex uptake was 20.2% and 45.8% lower in wild-type CHO-K1 compared to CHO-pgs-A745 for pHK10 and pHK15, respectively, although gene expression levels were similar between the two cell lines (Figure 5A) in contrast to observations seen with other lysine-based vectors.…”

Section: Resultsmentioning

confidence: 99%

Effect of Polyplex Morphology on Cellular Uptake, Intracellular Trafficking, and Transgene Expression

et al. 2013

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Nanoparticle morphology has been shown to affect cellular uptake, but there are few studies investigating the impact of particle shape on biologic drug delivery. Recently, our group synthesized a series of N-(2-hydroxypropyl) methacrylamide (HPMA)-oligolysine brush polymers for nucleic acid delivery that varied in oligolysine peptide length and polymer molecular weight. Interestingly, a 50% longer peptide (K15) transfected very poorly compared to the optimized polymer comprised of K10 peptide despite similar chemical composition and molecular weight. We hypothesized that differences in particle morphology contributed to the differences in plasmid DNA delivery. We found that particles formed with plasmid DNA and a polymer with the longer oligolysine peptide (pHK15) had larger aspect ratios than particles formed with optimized polymer (pHK10). Even though both formulations showed similar percentages of cellular association, particles of a higher aspect ratio were internalized to a lesser extent. Furthermore, the rod-like particles accumulated more in endosomal/lysosomal compartments, leading to delayed nuclear delivery. Other parameters, such as particle surface charge, unpackaging ability, uptake mechanism, intracellular trafficking, nor the presence of heparan sulfate proteoglycans did not significantly differ between the two polymer formulations. These results indicate that, for this system, polyplex morphology primarily impacts nucleic acid delivery efficiency through differences in cellular internalization rates.

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Section: Resultsmentioning

confidence: 99%

Effect of Polyplex Morphology on Cellular Uptake, Intracellular Trafficking, and Transgene Expression

et al. 2013

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“…Cell surface receptors such as the integrins 8, 9 and syndecans 10 have been shown to participate in gene transfer and interact with non-viral delivery systems. The interaction of cell surface glycosaminoglycans (GAGs) with non-viral vectors has also been shown to influence the intranuclear uptake and post nuclear processes 11 . Main parameters that are able to modulate gene transfer in 2-D include cell proliferation 12, 13 , internalization 14 and nuclear area 15 .…”

Section: Introductionmentioning

confidence: 99%

Transfection in the third dimension

Dhaliwal¹,

Oshita²,

Segura³

2013

Integr. Biol.

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An understanding of parameters that modulate gene transfer in 3-D will assist in the formation of gene delivery systems and scaffolds, which can mediate efficient non-viral delivery for guiding in-vivo tissue regeneration and therapy. We have previously demonstrated the cell area and length, integrin expression, and RhoGTPases mediated signalling to be pivotal parameters that guide gene transfer to mouse mesenchymal stem cells (mMSCs) cultured in 2-D and are modulated by ECM proteins. In this study, we were interested in determining if cationic polymer mediated gene transfer to cells seeded in 3-D would occur through different mechanisms as compared to 2-D. In particular, we examined the endocytosis pathways used to internalize polyplexes, and the role of cytoskeletal dynamics and RhoGTPases on non-viral gene transfer for cells seeded in 2-D and 3-D. Inhibition of clathrin- and caveolae- mediated endocytosis resulted in more drastic decrease in overall transgene expression for cells seeded in 3-D than those in 2-D. In addition, polyplex internalization was only significantly decreased in 3-D when clathrin-mediated endocytosis was inhibited, while caveolae-mediated endocytosis inhibition for cells seeded in 2-D resulted in the strongest polyplex internalization inhibition. Actin and microtubule polymerization affected 2-D and 3-D transfection differently. Microtubule depolymerization enhanced transgene expression in 2-D, but inhibited transgene expression in 3-D. Last, inhibition of RhoGTPases also affected 2-D and 3-D transfection differently. The inhibition of ROCK effector resulted in a decrease of transgene expression and internalization for cells seeded in 3-D, but not 2-D and the inhibition of effector PAK1 resulted in an increase of transgene expression for both 2-D and 3-D. Overall, our study suggests that the process of gene transfer occurs through different mechanisms for cells seeded in 2-D compared to those seeded in 3-D.

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“…In addition, the electrostatic compaction of DNA by dendrimers was found to protect it from degradation by nucleases and to enhance the cellular uptake of dendrimer-based DNA-containing particles via adsorptive endocytosis or phagocytosis. Polyamidoamine (PAMAM) dendrimers are commercially available macromolecules that can be used as carriers for plasmid DNA, antisense oligonucleotides, and siRNA [5][6][7][8]. However, PAMAM dendrimers are not biodegradable and exhibit strong cytotoxicity [9,10].…”

Section: Introductionmentioning

confidence: 99%

Biodegradable nanoparticles composed of dendrigraft poly-l-lysine for gene delivery

Kodama

Nakamura

Kurosaki

et al. 2014

European Journal of Pharmaceutics and Biopharmaceutics

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We developed novel gene vectors composed of dendrigraft poly-L-lysine (DGL).The transgene expression efficiency of the pDNA/DGL complexes (DGL complexes) was markedly higher than that of the control pDNA/poly-L-lysine complex. However, the DGL complexes caused cytotoxicity and erythrocyte agglutination at high doses.Therefore, γ-polyglutamic acid (γ-PGA), which is a biodegradable anionic polymer, was added to the DGL complexes to decrease their toxicity. The resultant ternary complexes (DGL/γ-PGA complexes) were shown to be stable nanoparticles, and those with γ-PGA to pDNA charge ratios of >8 had anionic surface charges. The transgene expression efficiency of the DGL/γ-PGA complexes was similar to that of the DGL complexes; however, they exhibited lower cytotoxicity and did not induce erythrocyte agglutination at high doses. After being intravenously administered to mice, the DGL6 complex demonstrated high transfection efficiency in the liver, lungs, and spleen, whereas the DGL6/γ-PGA8 complex only displayed high transfection efficiency in the spleen.

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Cell-surface glycosaminoglycans inhibit intranuclear uptake but promote post-nuclear processes of polyamidoamine dendrimer–pDNA transfection

Cited by 16 publications

References 44 publications

Effect of Polyplex Morphology on Cellular Uptake, Intracellular Trafficking, and Transgene Expression

Effect of Polyplex Morphology on Cellular Uptake, Intracellular Trafficking, and Transgene Expression

Transfection in the third dimension

Biodegradable nanoparticles composed of dendrigraft poly-l-lysine for gene delivery

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