Elucidating the pre- and post-nuclear intracellular processing of 1,4-dihydropyridine based gene delivery carriers

Hyvönen, Zanna; Hämäläinen, Vesa; Ruponen, Marika; Lucas, Bart; Rejman, Joanna; Vercauteren, Dries; Demeester, Jo; Smedt, Stefaan C. De; Braeckmans, Kevin

doi:10.1016/j.jconrel.2012.06.013

Cited by 16 publications

(22 citation statements)

References 61 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…As naked nucleic acids are negatively charged, they are complexed with cationic polymers or cationic liposomes to increase the cellular delivery. These complexes can take different endocytic pathways to enter the cells, dependent on their size and surface characteristics (11)(12)(13)(14). When nanoparticles fail to escape the endosomal compartment, eventual delivery to (and degradation in) the lysosomes will occur.…”

Section: Introductionmentioning

confidence: 99%

Lysosomal capturing of cytoplasmic injected nanoparticles by autophagy: An additional barrier to non viral gene delivery

Remaut

Oorschot²,

Braeckmans

et al. 2014

Journal of Controlled Release

Self Cite

View full text Add to dashboard Cite

Autophagy or 'self-eating' is a process by which defective organelles and foreign material can be cleared from the cell's cytoplasm and delivered to the lysosomes in which degradation occurs. It remains an open question, however, whether nanoparticles that did not enter the cell through endocytosis can also be captured from the cytoplasm by autophagy. We demonstrate that nanoparticles that are introduced directly in the cytoplasm of the cells by microinjection, can trigger an autophagy response. Moreover, both polystyrene beads and plasmid DNA containing poly-ethylene-imine complexes colocalize with autophagosomes and lysosomes, as was confirmed by electron microscopy.This indicates that cytoplasmic capturing of nanoparticles can occur by an autophagy response. The capturing of nanoparticles from the cytoplasm most likely limits the time frame in which efficient nucleic acid delivery can be obtained. Hence, autophagy forms an additional barrier to non-viral gene delivery, a notion that was not often taken into account before. Furthermore, these findings urges us to reconsider the idea that a single endosomal escape event is sufficient to have the long-lasting presence of nanoparticles in the cytoplasm of the cells.

show abstract

Section: Introductionmentioning

confidence: 99%

Lysosomal capturing of cytoplasmic injected nanoparticles by autophagy: An additional barrier to non viral gene delivery

Remaut

Oorschot²,

Braeckmans

et al. 2014

Journal of Controlled Release

Self Cite

View full text Add to dashboard Cite

show abstract

“…The difference in endosomal escape between the normal and the pHirrelevant scenario is then evaluated using the different endosomal escape assays which will be discussed further, and have been summarized in Figure 1. Removing the influence of pH on endosomal escape is done by either altering the compound under investigation so it is no longer pH-reactive [44], or the pH of the endosomal compartment itself is altered. In a controlled ex cellulo environment, the pH can be modeled by using buffers with different pH [45,46].…”

Section: Verifying Ph-induced Membrane Destabilizationmentioning

confidence: 99%

“…However, upon dye dilution over a larger surface area, these dimers will break apart, leading to a loss in fluorescence [59]. A more frequently used technique to monitor fluorophore dilution is dye [17,44,68]. Liposomal particles are double-labeled in a high concentration so that the distance between donor and acceptor allow FRET.…”

Section: Assays For Studying Membrane Fusionmentioning

confidence: 99%

“…Upon successful lipid fusion however, both fluorophores will be diluted over a larger surface area and the distance between donor and acceptor will increase, resulting in a decreased FRET efficiency. In an ex cellulo setting, this double-labeling can be applied to either the liposomal vector [17] or the artificial endosome [44], and FRET efficiency can be monitored by spectrofluorimetry. FRET has been used in cellular experiments as well, where the donor and acceptor fluorophores are incorporated in the lipid envelop of the nanoparticulate cargo and FRET efficiency is monitored by live cell spectral imaging microscopy [68].…”

Section: Assays For Studying Membrane Fusionmentioning

confidence: 99%

“…Fluorophore dilution in lipid membranes is frequently used to investigate membrane fusion, either induced by CPPs [55,59,68], viral nanoparticles [69][70][71] or liposomal delivery vectors of proteins [17] or pDNA [44,68]. Nevertheless, it should be emphasized that the major limitation of this assay is the inability to distinguish between lipid fusion and lipid mixing ( Figure 5A) [72].Whereas both fusion and mixing denotes the interaction between lipid bilayers and will lead to dilution of the incorporated fluorophores over a larger surface area, lipid mixing will not result in the intended displacement of the cargo.…”

Section: Assays For Studying Membrane Fusionmentioning

confidence: 99%

See 2 more Smart Citations

Intracellular delivery of nanomaterials: How to catch endosomal escape in the act

et al. 2014

Self Cite

View full text Add to dashboard Cite

Successful cytosolic delivery of nanomaterials is becoming more and more important, given the increase in intracellular applications of quantum dots, gold nanoparticles, liposomal drug formulations and polymeric gene delivery vectors. Most nanomaterials are taken up by the cell via endocytosis, yet endosomal escape has long been recognized as a major bottleneck in cytosolic delivery. Although it is essential to detect and reliably quantify endosomal escape, no consensus has been reached so far on the methods to do so. This review will summarize and discuss for the first time the different assays used to investigate this elusive step to date.

show abstract

Synthetic vectors for gene delivery: An overview of their evolution depending on routes of administration.

Belmadi

Midoux

Loyer

et al. 2015

Biotechnology Journal

View full text Add to dashboard Cite

Nucleic acid delivery constitutes an emerging therapeutic strategy to cure various human pathologies. This therapy consists of introducing genetic material into the whole body or isolated cells to correct a cellular abnormality or disfunction. As with any drug, the main objective of nucleic acid delivery is to establish optimal balance between efficacy and tolerance. The methods of administration and the vectors used are selected depending on whether the goal of treatment is the production of an active protein; the replacement of a missing or inactive gene; or the combat of acquired diseases, such as cancer or AIDS. In that sense, synthetic vectors represent a valuable solution because they are well characterized, their structure can be fine tuned, and their potential toxicity can be reduced, since toxicity depends on the composition of the formulations. Here we review various synthetic vectors for gene delivery and address the question of their biodistribution as a function of the route of administration. We highlight the modifications to vectors structure and formulations necessary to overcome the major hurdles limiting the effectiveness of nucleic acid therapies.

show abstract

Elucidating the pre- and post-nuclear intracellular processing of 1,4-dihydropyridine based gene delivery carriers

Cited by 16 publications

References 61 publications

Lysosomal capturing of cytoplasmic injected nanoparticles by autophagy: An additional barrier to non viral gene delivery

Lysosomal capturing of cytoplasmic injected nanoparticles by autophagy: An additional barrier to non viral gene delivery

Intracellular delivery of nanomaterials: How to catch endosomal escape in the act

Synthetic vectors for gene delivery: An overview of their evolution depending on routes of administration.

Contact Info

Product

Resources

About