Non-linear pharmacokinetics of octaarginine-modified lipid nanoparticles: Barriers from in vitro to in vivo

Hayashi, Yasuhiko; Noguchi, Yuki; Harashima, Hideyoshi

doi:10.1016/j.jconrel.2012.05.036

Cited by 11 publications

(5 citation statements)

References 19 publications

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“…Although the separation resolution was limited, some quantitative kinetics data could be extracted from the subcellular fractionation ( Figure 4B). The amount of siRNA in the heaviest fractions (35)(36)(37)(38)(39)(40)(41)(42)(43)(44)(45) and the lightest fractions (0-10) increased throughout the experiment .…”

Section: Discussionmentioning

confidence: 99%

“…We investigated the rates of PEI/AF647-siRNA polyplex attachment to the cell membrane and cellular internalization. Cationic gene delivery vectors bind anionic proteoglycans on the cell surface [42,43] and can be removed by washing the cells in the presence of heparin to compete for binding of polyplexes [34,44,45]. We transfected HeLa-luc cells with PEI/AF647-siRNA Figure 2A).…”

Section: Kinetics Of Pei/sirna Uptakementioning

confidence: 99%

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Rapid and facile quantitation of polyplex endocytic trafficking

Lazebnik

Pack

2017

Journal of Controlled Release

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Design of safe and effective synthetic nucleic acid delivery vectors such as polycation/DNA or polycation/siRNA complexes (polyplexes) will be facilitated by quantitative understanding of the mechanisms by which such materials escort cargo from the cell surface to the nucleus. In particular, the mechanisms of cellular internalization by various endocytosis pathways and subsequent endocytic vesicle trafficking have been shown to strongly affect nucleic acid delivery efficiency. Fluorescence microscopy and subcellular fractionation methods are commonly employed to follow intracellular trafficking of biomolecules and nanoparticulate delivery systems such as polyplexes. However, it is difficult to obtain quantitative data from microscopy and subcellular fractionation is experimentally difficult and low throughput. We have developed a method for quantifying the transport of polyplexes through important endocytic vesicles. The method is based on polymerization of 3,3'-diaminobenzidine by endocytosed horseradish peroxidase, causing an increase in the vesicle density, resistance to being solubilized by detergent and quenching of fluorophores within the vesicles, which makes them easy to separate and quantify. Using this method in HeLa cells, we have observed polyethylenimine/siRNA polyplexes initially appearing in early endosomes and rapidly moving to other compartments within 30min post-transfection. At the same time, we observed the kinetics of accumulation of the polyplexes in lysosomes at a similar rate. The results from the new method are consistent with similar measurements by confocal fluorescence microscopy and subcellular fractionation of endocytic vesicles on a Percoll gradient. The relative ease of this new method will aid investigation of gene delivery mechanisms by providing the means to rapidly quantify endocytic trafficking of polyplexes and other vectors.

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Section: Discussionmentioning

confidence: 99%

Section: Kinetics Of Pei/sirna Uptakementioning

confidence: 99%

Rapid and facile quantitation of polyplex endocytic trafficking

Lazebnik

Pack

2017

Journal of Controlled Release

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“…Pharmacokinetic (PK) and pharmacodynamic (PD) factors also need to be considered when using tumor cylindroids. Harashima and coworkers investigated the discrepancies between in vitro and in vivo systems in terms of different PKs and PDs using octaarginine (R8) modified engineered lipid NPs in mice for siRNA delivery [74]. These results showed a remarkable difference in the PKs between the two types of systems.…”

Section: Endogenous Activationmentioning

confidence: 99%

Triggered nanoparticles as therapeutics

et al. 2013

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Summary Drug delivery systems (DDSs) face several challenges including site-specific delivery, stability, and the programmed release of drugs. Engineered nanoparticle (NP) surfaces with responsive moieties can enhance the efficacy of DDSs for in vitro and in vivo systems. This triggering process can be achieved through both endogenous (biologically controlled release) and exogenous (external stimuli controlled release) activation. In this review, we will highlight recent examples of the use of triggered release strategies of engineered nanomaterials for in vitro and in vivo applications.

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“…Furthermore, siRNA degradation profile was similar in both cultured cells and the mouse liver. However, a remarkable nonlinearity was observed in PK as shown in Table , indicating that the percentage of siRNA amount detected in the mouse liver was drastically decreased as the treatment dose was decreased. These results demonstrated that the PK is a causative factor in the huge gap between the in vitro and in vivo situations, and these findings provide a promising clue to achieving a more efficient CPP-mediated in vivo siRNA delivery at a lower dose.…”

Section: R8-mend For In Vivo Deliverymentioning

confidence: 99%

Multifunctional Envelope-Type Nano Device: Evolution from Nonselective to Active Targeting System

et al. 2015

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A paradigm shift has occurred in the field of drug delivery systems (DDS), one being intracellular targeting, and the other, active targeting. An important aspect of intracellular targeting involves delivering nucleic acids such as siRNA/pDNA rather than small molecular compounds, since the mechanism responsible for their entering a target cell is usually via endocytosis, and the efficiency of endosomal escape is a critical factor in determining the functional activities of siRNA/pDNA. A multifunctional envelope-type nano device (MEND) was developed to control the intracellular trafficking of nano carriers containing siRNA/pDNA. An octaarginine (R8) modified MEND was developed to achieve this. Considerable progress has been made in active targeting to selective tissue vasculature such as tumor, adipose tissue, and the lung where endothelial barrier is tight against nanoparticles with diameters larger than 50 nm. A dual-ligand system is proposed to enhance active targeting ability by virtue of a synergistic interaction between a selective ligand and a cell penetrating ligand. Prohibitin targeted nanoparticles (PTNP) were developed to target endothelial cells in adipose tissue, which deliver apoptotic peptides/proteins to the adipose vasculature. Lung endothelial cells can be targeted by means of the GALA peptide, which is usually used to enhance endosomal escape. These active targeting systems can induce pharmacological effects in in vivo conditions. Finally, a novel strategy for producing an original ligand has been developed, especially for the tumor vasculature. This progress in DDS promises to extend the area of nanomedicine as a breakthrough technology.

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Non-linear pharmacokinetics of octaarginine-modified lipid nanoparticles: Barriers from in vitro to in vivo

Cited by 11 publications

References 19 publications

Rapid and facile quantitation of polyplex endocytic trafficking

Rapid and facile quantitation of polyplex endocytic trafficking

Triggered nanoparticles as therapeutics

Multifunctional Envelope-Type Nano Device: Evolution from Nonselective to Active Targeting System

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