Multifunctional envelope-type nano device (MEND) as a non-viral gene delivery system☆
Introduction
A successful gene delivery system requires a rational strategy for overcoming many biological barriers such as membrane and enzymatic barriers. When plasmid DNA (pDNA) packaged in a delivery system is internalized via endocytosis, it must escape from endosomes before lysosomal degradation. The pDNA also must pass through the nuclear membrane before it can be transcribed in the nucleus. Therefore, the ideal gene delivery system should be equipped with a variety of functional devices to overcome these barriers, such as ligands for specific targeting, pH-sensitive fusogenic peptides, and a nuclear localization signal (NLS) [1], [2], [3], [4], [5]. Most non-viral vectors developed to date contain some of these devices [1], [2], [3], [4], [5]; however, it is difficult to integrate all of them into a single delivery system and to have each function exerted according to a specific program to overcome the barriers. Simple mixing of these nano devices is not sufficient for developing such an “artificially intelligent” nano device. Therefore, a new concept was required to develop a non-viral gene delivery system which can compete with viral ones in terms of efficiency. We succeeded in developing a multifunctional envelope-type nano device (MEND) as a non-viral gene delivery system based on a new concept, “Programmed Packaging”. In this review, we describe how we developed this concept and how we construct the MEND by focusing on the function of octaarginine as a key molecular component of the MEND.
Section snippets
Importance of topology control
Since the R8 peptide was found to be taken up via a nonclassical endocytic pathway [6], which can circumvent lysosomal degradation, R8 was proposed as a functional device for efficient intracellular trafficking of nanoparticles, such as liposomes. Thus, the mechanism of uptake of nanoparticles tagged with R8 peptide was confirmed using nano-size complexes of pDNA encoding luciferase and complexed with either R8 peptide or stearylated R8 (STR-R8) [7]. The transfection activity of the STR-R8/DNA
A new concept: programmed packaging
Recently, a novel non-viral gene delivery system was developed based on a new packaging concept termed “Programmed Packaging”, and was named the MEND [17]. Programmed Packaging was proposed to develop rational non-viral gene delivery systems equipped with various functional devices, including ligands for specific receptors, pH-sensitive fusogenic peptides for endosomal escape, and a NLS for enhanced nuclear delivery [5] to overcome several barriers in the process of gene delivery to the nucleus
MEND for oligonucleotides
The R8-MEND was suggested to be a useful delivery system for antisense oligodeoxynucleotides (ODN), which can inhibit the function of mRNA in the cytosolic space, since R8-MEND showed high efficacy in pDNA delivery, as mentioned above. For delivery of ODN, ODN-encapsulated R8-MEND (ODN-MEND) was developed [20], [21]. Three types of R8-MENDs were constructed by co-encapsulation of luciferase-encoding pDNA and anti-luciferase ODN condensed by three polycations – STR-R8, PLL and protamine – and
Drug carriers for mitochondrial drug therapy
Mitochondrial dysfunction has recently been implicated in a variety of diseases, including neurodegenerative and neuromuscular disorders, obesity and diabetes mellitus, ischemia-reperfusion injury, cancer and inherited mitochondrial diseases [25], [26]. Although therapies for these diseases are desired, strategies designed to complement mitochondrial dysfunction are rare [27], [28]. To enable drug delivery targeted to mitochondria, we first need to establish a method to encapsulate various
Quantitative analytical method CIDIQ
As described above, we control the intracellular trafficking of pDNA, ODN, siRNA and proteins using nano particles developed based on the concept of Programmed Packaging. For the development of new-generation gene-delivery vectors, a more rational strategy is required, based on feedback information for intracellular trafficking. This information would enable us to better recognize which of the barriers need to be overcome to improve the gene-delivery activity. Recently, we proposed a novel
Conclusion
MEND has been developed based on the concept of Programmed Packaging and applied to not only pDNA but also to ODN, siRNA and other molecules. A comparative analysis between viral and non-viral gene vectors indicated that post-nuclear delivery process is the rate-limiting step in transfection mediated by non-viral gene delivery systems. Therefore, further efforts should be directed at optimizing the intranuclear disposition of MEND. At the same time, MEND can be extended not only to nuclear
Acknowledgement
The authors wish to thank Dr. Daryl Henderson for his helpful advice in writing the English manuscript.
References (53)
- et al.
Cellular and molecular barriers to gene transfer by a cationic lipid
J. Biol. Chem.
(1995) - et al.
Folate-targeted, anionic liposome-entrapped polylysine-condensed DNA for tumor cell-specific gene transfer
J. Biol. Chem.
(1996) - et al.
Pharmacokinetic and pharmacodynamic considerations in gene therapy
Drug Discov. Today
(2003) - et al.
Arginine-rich peptides. An abundant source of membrane-permeable peptides having potential as carriers for intracellular protein delivery
J. Biol. Chem.
(2001) - et al.
High density of octaarginine stimulates macropinocytosis leading to efficient intracellular trafficking for gene expression
J. Biol. Chem.
(2006) - et al.
Cellular uptake and subsequent intracellular trafficking of R8-liposomes introduced at low temperature
Biochim. Biophys. Acta
(2006) - et al.
Cellular uptake of arginine-rich peptides: roles for macropinocytosis and actin rearrangement
Mol. Ther.
(2004) - et al.
Protein transduction domain of HIV-1 Tat protein promotes efficient delivery of DNA into mammalian cells
J. Biol. Chem.
(2001) - et al.
Cell-penetrating peptides. A reevaluation of the mechanism of cellular uptake
J. Biol. Chem.
(2003) - et al.
Uptake of analogs of penetratin, Tat(48–60) and oligoarginine in live cells
Biochem. Biophys. Res. Commun.
(2003)
Development of a non-viral multifunctional envelope-type nano device by a novel lipid film hydration method
J. Control. Release
A multifunctional envelope-type nano device for novel gene delivery of siRNA plasmids
Int. J. Pharm.
Octaarginine-modified multifunctional envelope-type nano device for siRNA
J. Control. Release
New packaging method of mycobacterial cell wall using octaarginine-modified liposomes: enhanced uptake by and immunostimulatory activity of dendritic cells
J. Control. Release
Mitochondria: dynamic organelles in disease, aging, and development
Cell
Mitochondrial disease
Lancet
Mitochondrial pharmaceutics
Mitochondrion
Mitochondrial drug delivery and mitochondrial disease therapy — an approach to liposome-based delivery targeted to mitochondria
Mitochondrion
Nuclear genes encoding mitochondrial proteins in yeast
Trends Biochem. Sci
The protein import system of mitochondria
J. Biol. Chem.
Targeted delivery of DNA to the mitochondrial compartment via import sequence-conjugated peptide nucleic acid
Mol. Ther.
Expression of human ferredoxin in Saccharomyces cerevisiae: mitochondrial import of the protein and assembly of the [2Fe–2S] center
Arch. Biochem. Biophys.
Photosensitisation properties of mitochondrially localised green fluorescent protein
Biochem. Biophys. Res. Commun.
Protein transduction technology
Curr. Opin. Biotechnol.
Intracellular delivery of large molecules and small particles by cell-penetrating proteins and peptides
Adv. Drug Deliv. Rev.
A novel TAT-mitochondrial signal sequence fusion protein is processed, stays in mitochondria, and crosses the placenta
Mol. Ther.
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This review is part of the Advanced Drug Delivery Reviews theme issue on “Membrane Permeable Peptide Vectors: Chemistry and Functional Design for the Therapeutic Applications”.
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Present address: Kyoto Pharmaceutical University, Japan.