Vitreous: A Barrier to Nonviral Ocular Gene Therapy

Peeters, Liesbeth; Sanders, Niek N.; Braeckmans, Kevin; Boussery, Koen; Voorde, Johan Van de; Smedt, Stefaan C. De; Demeester, Joseph

doi:10.1167/iovs.05-0165

Cited by 175 publications

(139 citation statements)

References 39 publications

Supporting

Mentioning

123

Contrasting

Unclassified

Order By: Relevance

“…In animal models, an in vitro-in vivo correlation is not necessarily expected, because of possible physical barriers (Peeters et al, 2005;de la Fuente et al, 2010).…”

Section: Discussionmentioning

confidence: 99%

Dextran and Protamine-Based Solid Lipid Nanoparticles as Potential Vectors for the Treatment of X-Linked Juvenile Retinoschisis

et al. 2012

View full text Add to dashboard Cite

The goal of the present study was to analyze the potential application of nonviral vectors based on solid lipid nanoparticles (SLN) for the treatment of ocular diseases by gene therapy, specifically X-linked juvenile retinoschisis (XLRS). Vectors were prepared with SLN, dextran, protamine, and a plasmid (pCMS-EGFP or pCEP4-RS1). Formulations were characterized and the in vitro transfection capacity as well as the cellular uptake and the intracellular trafficking were studied in ARPE-19 cells. Formulations were also tested in vivo in Wistar rat eyes, and the efficacy was studied by monitoring the expression of enhanced green fluorescent protein (EGFP) after intravitreal, subretinal, and topical administration. The presence of dextran and protamine in the SLN improved greatly the expression of retinoschisin and EGFP in ARPE-19 cells. The nuclear localization signals of protamine, its ability to protect the DNA, and a shift in the entry mechanism from caveola-mediated to clathrinmediated endocytosis promoted by the dextran, justify the increase in transfection. After ocular administration of the dextran-protamine-DNA-SLN complex to rat eyes, we detected the expression of EGFP in various types of cells depending on the administration route. Our vectors were also able to transfect corneal cells after topical application. We have demonstrated the potential usefulness of our nonviral vectors loaded with XLRS1 plasmid and provided evidence for their potential application for the management or treatment of degenerative retinal disorders as well as ocular surface diseases.

show abstract

“…In animal models, an in vitro-in vivo correlation is not necessarily expected, because of possible physical barriers (Peeters et al, 2005;de la Fuente et al, 2010).…”

Section: Discussionmentioning

confidence: 99%

Dextran and Protamine-Based Solid Lipid Nanoparticles as Potential Vectors for the Treatment of X-Linked Juvenile Retinoschisis

et al. 2012

View full text Add to dashboard Cite

show abstract

“…Polystyrene beads 30 and positively charged beads (see Table 1). The latter is the result of an amide formation between the carboxylate groups on the beads and the amine group of DMAEA.…”

Section: Methodsmentioning

confidence: 99%

Nuclear Inclusion of Nontargeted and Chromatin-Targeted Polystyrene Beads and Plasmid DNA Containing Nanoparticles

et al. 2011

Self Cite

View full text Add to dashboard Cite

ABSTRACT. The nuclear membrane is one of the major cellular barriers in the delivery of plasmid DNA (pDNA). Cell division has a positive influence on the expression efficiency since at the end of mitosis, pDNA or pDNA containing complexes near the chromatin are probably included by a random process in the nuclei of the daughter cells. However, very little is known about the nuclear inclusion of nanoparticles during cell division. Using the Xenopus nuclear envelope reassembly (XNER) assay, we found that the nuclear enclosure of nanoparticles was dependent on size (with 100 nm and 200 nm particles being better included than the 500 nm ones) and charge (with positively charged particles being better included than negatively charged or poly-ethyleneglycolated (PEG-ylated) ones) of the 2 beads. Also, coupling chromatin-targeting peptides to the polystyrene beads or pDNA complexes improved their inclusion by 2-to 3-fold. Upon microinjection in living HeLa cells, however, nanoparticles were never observed in the nuclei of cells post-division but accumulated in a specific perinuclear region, which was identified as the lysosomal compartment. This indicates that nanoparticles can end up in the lysosomes even when they were not delivered through endocytosis. To elucidate if the chromatin binding peptides also have potential in living cells, this additional barrier first has to be tackled, since it prevents free particles to be present near the chromatin at the moment of cell division.

show abstract

“…14,15) The cationic complexes, however, cause severe cytotoxicity in retinal pigment epithelial cells and aggregation with the vitreous body in case of intravitreal injection because of their strong cationic surface charge. [16][17][18][19] These side effects could decrease visual acuity and even cause blindness.…”

mentioning

confidence: 99%

Ocular Gene Delivery Systems Using Ternary Complexes of Plasmid DNA, Polyethylenimine, and Anionic Polymers

Kurosaki

Uematsu

Shimoda

et al. 2013

Biological & Pharmaceutical Bulletin

View full text Add to dashboard Cite

In this experiment, we developed anionic ternary complexes for effective and safe ocular gene delivery. Ternary complexes were constructed by coating plasmid DNA (pDNA)/polyethylenimine (PEI) complex with anionic polymers such as γ-polyglutamic acid (γ-PGA) and chondroitin sulfate (CS). The cationic pDNA/PEI complex showed high gene expression on the human retinal pigment epithelial cell line, ARPE-19 cells. The pDNA/PEI complexes, however, also showed high cytotoxicity on the cells and aggregated strongly in the vitreous body. On the other hand, the anionic ternary complexes showed high gene expression on ARPE-19 cells without such cytotoxicity and aggregation. After intravitreous administration of the complexes, the anionic ternary complexes showed high gene expression in the retina. These results strongly indicate that anionic ternary complexes are suitable for effective and safe ocular gene therapy.

show abstract

Vitreous: A Barrier to Nonviral Ocular Gene Therapy

Abstract: Modifying the surfaces of LPXs with hydrophilic PEG chains prevents them from aggregating in vitreous. In this way, LPXs are obtained that can freely move in vitreous, an absolute criterion for reaching the retina after intravitreal injection.

Cited by 175 publications

References 39 publications

Dextran and Protamine-Based Solid Lipid Nanoparticles as Potential Vectors for the Treatment of X-Linked Juvenile Retinoschisis

Dextran and Protamine-Based Solid Lipid Nanoparticles as Potential Vectors for the Treatment of X-Linked Juvenile Retinoschisis

Nuclear Inclusion of Nontargeted and Chromatin-Targeted Polystyrene Beads and Plasmid DNA Containing Nanoparticles

Ocular Gene Delivery Systems Using Ternary Complexes of Plasmid DNA, Polyethylenimine, and Anionic Polymers

Contact Info

Product

Resources

About