Potocytosis and cellular exit of complexes as cellular pathways for gene delivery by polycations

Grosse, Stéphanie; Aron, Y.; Thévenot, Guiti; François, Dominique; Monsigny, Michel; Fajac, Isabelle

doi:10.1002/jgm.772

Cited by 92 publications

(71 citation statements)

References 36 publications

(55 reference statements)

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“…The results of the present study are consistent with prior studies showing that large latex beads preferentially enter via caveolar endocytosis, although the mechanism is unclear [24,25]. However, other studies showed that particles with sizes exceeding 200 nm were internalized via macropinocytosis rather than caveolar endocytosis [26]. Taken together, these results suggest that the preferred cellular uptake pathway of IRQ-Lip particles could not be simply discussed by its physicochemical properties such as charge and size.…”

Section: Resultssupporting

confidence: 91%

Topology of octaarginines (R8) or IRQ ligand on liposomes affects the contribution of macropinocytosis- and caveolae-mediated cellular uptake

Mudhakir¹,

Akita²,

Harashima³

2011

Reactive and Functional Polymers

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It was recently reported that liposomes modified with octaarginine (R8) and its analogue peptide (IRQRRRR: IRQ) are taken into NIH3T3 cells by unique pathways, macropinocytosis and caveolae-mediated endocytosis, respectively. This study evaluated the topology of these peptides as it relates to the uptake routes of liposomes, where they are modified either directly on the surface, or on the edge of a polyethylene glycol (PEG) spacer. The uptake mechanism of peptide-modified liposomes and peptide-modified PEG-liposomes was investigated by confocal laser scanning microscopy. To determine the contribution of clathrin-mediated endocytosis, macropinocytosis and caveolar endocytosis to the uptake of liposomes, the uptake was evaluated in the presence of these specific inhibitors. The uptake pathway changed from macropinocytosis to clathrin-mediated endocytosis when R8 was modified on the edge of a PEG spacer, indicating that the flexible display of R8 impaired the induction of macropinocytosis. However, the contribution of caveolae-mediated endocytosis increased when IRQ was conjugated to the distal end of the PEG chain, suggesting that flexible surface display enhanced IRQ recognition by the specific molecules in the caveolae. The present results demonstrate that topology control of the ligand affects the contribution of the entry pathway, depending on the uptake mechanism.

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Section: Resultssupporting

confidence: 91%

Topology of octaarginines (R8) or IRQ ligand on liposomes affects the contribution of macropinocytosis- and caveolae-mediated cellular uptake

Mudhakir¹,

Akita²,

Harashima³

2011

Reactive and Functional Polymers

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“…Lanes 3-14: polyplexes of pEGFP (0.2 μg) with G6 (lanes 3, 4, 5), G6-4% PEG (lanes 6, 7, 8), G6-8% PEG (lanes 9, 10, 11), or G6-15% PEG (lanes 12, 13, 14). Each kind of dendrimer was used to form polyplexes with pEGFP in serum-free DMEM at three different N/P ratios of 1, 5, and 10, respectively dominant internalization pathway for polyplexes reported in the literature (39)(40)(41).…”

Section: Size and ζ Potential Of Peg-pamam Dendrimer/dna Polyplexesmentioning

confidence: 99%

PEG-conjugated PAMAM Dendrimers Mediate Efficient Intramuscular Gene Expression

Gao

Tang

et al. 2009

AAPS J

152

103

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Abstract. Generations 5 and 6 (G5 and G6) poly(amidoamine) (PAMAM) dendrimers have been shown to be highly efficient nonviral carriers in in vitro gene delivery. However, their high toxicity and unsatisfied in vivo efficacy limit their applications. In this study, to improve their characteristics as gene delivery carriers, polyethylene glycol (PEG, molecular weight 5,000) was conjugated to G5 and G6 PAMAM dendrimers (PEG-PAMAM) at three different molar ratios of 4%, 8%, and 15% (PEG to surface amine per PAMAM dendrimer molecular). Compared with unconjugated PAMAM dendrimers, PEG conjugation significantly decreased the in vitro and in vivo cytotoxicities and hemolysis of G5 and G6 dendrimers, especially at higher PEG molar ratios. Among all of the PEG-PAMAM dendrimers, 8% PEG-conjugated G5 and G6 dendrimers (G5-8% PEG, G6-8% PEG) resulted in the most efficient muscular gene expression when polyplexes were injected intramuscularly to the quadriceps of neonatal mice. Consistent with the in vivo results, these two 8% PEG-conjugated PAMAM dendrimers could also mediate the highest in vitro transfection in 293A cells. Therefore, G5-8% PEG and G6-8% PEG possess a great potential for gene delivery both in vivo and in vitro.

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“…In CLSM analyses of HUVEC and VSMC, pDNA transferred with cRGD-PEG-PAsp(DET) micelles tended to colocalize with lipid rafts and caveosomes, whereas pDNA after transfer with PEG-PAsp(DET) micelles tended to associate with acidic endosomes and lysosomes. Grosse et al 38 evaluated the cellular uptake of particles that were not installed with any ligands and showed that the process of cellular uptake depended upon particle size; particles with 4200 nm diameter were internalized by macropinocytosis, particles with 100-200 nm diameter by clathrin-mediated endocytosis and particles o100 nm diameter by caveolae-mediated endocytosis. The average diameter of cRGD-PEG-PAsp(DET) and PEG-PAsp(DET) micelles was around 100 nm, with a moderate distribution.…”

Section: Discussionmentioning

confidence: 99%

Impact of polyplex micelles installed with cyclic RGD peptide as ligand on gene delivery to vascular lesions

et al. 2011

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Gene therapy is expected to open a new strategy for the treatment of refractory vascular diseases, so the development of appropriate gene vectors for vascular lesions is needed. To realize this requirement with a non-viral approach, cyclo(RGDfK) peptide (cRGD) was introduced to block copolymer, poly(ethylene glycol)-block-polycation carrying ethylenediamine units (PEG-PAsp(DET)). cRGD recognizes a v b 3 and a v b 5 integrins, which are abundantly expressed in vascular lesions. cRGDconjugated PEG-PAsp(DET) (cRGD-PEG-PAsp(DET)) formed polyplex micelles through complexation with plasmid DNA (pDNA) and the cRGD-PEG-PAsp(DET) micelles achieved significantly more efficient gene expression and cellular uptake as compared with PEG-PAsp(DET) micelles in endothelial cells and vascular smooth muscle cells. Intracellular tracking of pDNA showed that cRGD-PEG-PAsp(DET) micelles were internalized via caveolae-mediated endocytosis, which is associated with a pathway avoiding lysosomal degradation and that, PEG-PAsp(DET) micelles were transported to acidic endosomes and lysosomes via clathrin-mediated endocytosis. Further, in vivo evaluation in rat carotid artery with a neointimal lesion revealed that cRGD-PEGPAsp(DET) micelles realized sustained gene expression, whereas PEG-PAsp(DET) micelles facilitated rapid, but transient gene expression. These findings suggest that introduction of cRGD to polyplex micelles might create novel and useful functions for gene transfer and contribute to the establishment of efficient gene therapy for vascular diseases.

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Potocytosis and cellular exit of complexes as cellular pathways for gene delivery by polycations

Cited by 92 publications

References 36 publications

Topology of octaarginines (R8) or IRQ ligand on liposomes affects the contribution of macropinocytosis- and caveolae-mediated cellular uptake

Topology of octaarginines (R8) or IRQ ligand on liposomes affects the contribution of macropinocytosis- and caveolae-mediated cellular uptake

PEG-conjugated PAMAM Dendrimers Mediate Efficient Intramuscular Gene Expression

Impact of polyplex micelles installed with cyclic RGD peptide as ligand on gene delivery to vascular lesions

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