Gene delivery using dimethyldidodecylammonium bromide-coated PLGA nanoparticles
Introduction
The application of nucleic acids as therapeutic agents for gene therapy has been extensively studied in a broad range of diseases [1], [2], [3], [4]. However, a recurrent limitation in these therapies is the efficient delivery of the therapeutic DNA to the disease site. To address this issue, various strategies have been examined including vectors engineered from adeno- or adeno-associated viruses [5], but clinical trials have demonstrated substantial obstacles to their use, such as immunogenicity and inflammatory potential [6].
An alternative strategy is the application of non-viral gene delivery vectors, including liposomes [7], dendrimers [8], polycationic polymers [9], [10] and polymeric nanoparticles (NP) [11] to reduce or avoid immunogenicity and associated risks of toxicity [12]. A frequently employed biodegradable polymer in NP formulation is poly(lactic-co-glycolic acid) (PLGA). PLGA NP have shown particular promise in the delivery of a range of drug molecules to disease sites to improve efficacies [13], [14], [15], [16]. Moreover, the non-toxic biodegradability of PLGA has resulted in FDA approval for the application of this polymer in various medicinal products [17].
One approach in the application of PLGA NP for nucleic acid delivery uses adsorption of the anionic DNA molecules onto cationic NP [18], [19]. Despite this efficient formulation procedure, the peripheral exposure of the labile DNA limits stability and shortens its half-life, particularly in the acidic environment found within late endosomes where the particles will accumulate upon internalisation [20], [21]. Therefore, formulations that can encapsulate and protect the DNA from degradation are attractive for gene therapy approaches.
In addition to the protection of the DNA cargo, the physical characteristics of the nanoparticles can be manipulated to escape the degradative endosomal lumen, resulting in cytosolic localisation. Various strategies have been used to promote this sub-cellular relocalisation including application of fusogenic peptides [22], proton sponge polymers [23], light excitation [24] and cationic coating [18], [19], [25], [26]. It is thought that the presence of a cationic surface charge promotes interaction and binding of the NP to the endosomal membrane, inducing membrane destabilisation and cytosolic relocalisation of the NP [27].On this basis, the objective of this current study was to develop a PLGA NP formulation that would combine the ability to produce NP encapsulating DNA with the capacity to evade endosomal degradation.
Section snippets
Materials
PLGA (Resomer RG 502 H) with an acid value of 9 mg KOH per g PLGA, molecular weight 12 kDa, was a generous gift from Boehringer Ingelheim, Germany. Poly(vinyl alcohol) (PVA), 87–89% hydrolysed with molecular weight 13–23 kDa, and dimethyldidodecylammonium bromide (DMAB) were obtained from Sigma Aldrich, Germany. Magnesium chloride hexahydrate (MgCl2·6H2O), sodium bicarbonate NaHCO3, Tris-EDTA buffer and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT reagent) were obtained
Nanoparticle formulation
Upon internalisation by endocytosis, NP preferentially localise to late endosomes [20]. This localisation can be destructive to cargo DNA, particularly if adsorbed to the surface of cationic NP. Premature dissociation of surface-adsorbed DNA from NP in endosomal compartments following cellular internalisation has previously been observed [21], where degradation occurs rapidly in the acidified environment. Thus, the ability to entrap the nucleic acid inside the PLGA NP is attractive in order to
Conclusion
In summary, the application of a salting out and emulsion–evaporation process has successfully resulted in the formulation of 240 nm DNA-loaded NP. The introduction of a cationic surfactant (DMAB) has facilitated the endosomal escape of endocytosed NP, thus preventing premature degradation of the transfected DNA. This effect was exemplified by the observation of improved transfection efficiencies over anionic NP carrying a reporter plasmid. These DNA delivery modalities require approximately
References (65)
- et al.
Antigen delivery systems for veterinary vaccine development. Viral-vector based delivery systems
Vaccine
(2008) - et al.
Chitosan-coated PLGA nanoparticles for DNA/RNA delivery: effect of the formulation parameters on complexation and transfection of antisense oligonucleotides
Nanomedicine
(2007) - et al.
Ultradeformable cationic liposomes for delivery of small interfering RNA (siRNA) into human primary melanocytes
J Control Release
(2009) - et al.
Current status of polymeric gene delivery systems
Adv Drug Deliv Rev
(2006) - et al.
Biodegradable nanoparticles modified by branched polyethylenimine for plasmid DNA delivery
Biomaterials
(2010) - et al.
High loading efficiency and tunable release of plasmid DNA encapsulated in submicron particles fabricated from PLGA conjugated with poly-l-lysine
J Control Release
(2008) - et al.
Preparation and characterization of PE38KDEL-loaded anti-HER2 nanoparticles for targeted cancer therapy
J Control Release
(2008) - et al.
Targeting cancer cells using PLGA nanoparticles surface modified with monoclonal antibody
J Control Release
(2007) - et al.
PEGylated PLGA-based nanoparticles targeting M cells for oral vaccination
J Control Release
(2007) - et al.
Physicochemical characterization of poly(l-lactic acid) and poly(d, l-lactide-co-glycolide) nanoparticles with polyethylenimine as gene delivery carrier
Int J Pharm
(2005)
Preparation, characterization, cytotoxicity and transfection efficiency of poly(dl-lactide-co-glycolide) and poly(dl-lactic acid) cationic nanoparticles for controlled delivery of plasmid DNA
Int J Pharm
Fluorescence and electron microscopy probes for cellular and tissue uptake of poly(d, l-lactide-co-glycolide) nanoparticles
Int J Pharm
In vivo gene delivery in the mouse lung with lactosylated polyethylenimine, questioning the relevance of in vitro experiments
J Control Release
Fusogenic peptides enhance endosomal escape improving siRNA-induced silencing of oncogenes
Int J Pharm
Non-toxic phototriggered gene transfection by PAMAM-porphyrin conjugates
J Control Release
Design of biodegradable particles for protein delivery
J Control Release
A modified protocol for efficient DNA encapsulation into pegylated immunoliposomes (PILs)
J Control Release
In vivo gene gun-mediated DNA delivery into rodent brain tissue
Biochem Biophys Res Commun
Stability of peptide-condensed plasmid DNA formulations
J Pharm Sci
Microencapsulation of DNA using poly(Image-lactide-co-glycolide): stability issues and release characteristics
J Control Release
Properties of poly(lactic-co-glycolic acid) nanospheres containing protease inhibitors: camostat mesilate and nafamostat mesilate
Int J Pharm
Preservation of lysozyme structure and function upon encapsulation and release from poly(lactic-co-glycolic) acid microspheres prepared by the water-in-oil-in-water method
Int J Pharm
Transfection of a mouse dendritic cell line by plasmid DNA-loaded PLGA microparticles in vitro
Eur J Pharm Biopharm
Cationic microparticles consisting of poly(lactide-co-glycolide) and polyethylenimine as carriers systems for parental DNA vaccination
J Control Release
Augmented humoral and cellular immune responses to hepatitis B DNA vaccine adsorbed onto cationic microparticles
J Control Release
Stable cationic microparticles for enhanced model antigen delivery to dendritic cells
J Control Release
Preparation and purification of cationic solid lipid nanospheres-effects on particle size, physical stability and cell toxicity
Int J Pharm
Direct plasmid DNA encapsulation within PLGA nanospheres by single oil-in-water emulsion method
Eur J Pharm Biopharm
Formulation of functionalized PLGA-PEG nanoparticles for in vivo targeted drug delivery
Biomaterials
Reduction in burst release of PLGA microparticles by incorporation into cubic phase-forming systems
Eur J Pharm Biopharm
Selective disruption of lysosomes in HeLa cells triggers apoptosis mediated by cleavage of Bid by multiple papain-like lysosomal cathepsins
J Biol Chem
Demethylation using the epigenetic modifier, 5-azacytidine, increases the efficiency of transient transfection of macrophages
J Lipid Res
Cited by (55)
Gene therapy using PLGA nanoparticles
2023, Poly(lactic-co-glycolic acid) (PLGA) Nanoparticles for Drug DeliveryNanomedicine-based delivery strategies for nucleic acid gene inhibitors in inflammatory diseases
2021, Advanced Drug Delivery ReviewsCitation Excerpt :However, it must be noted that the sensitivities of a specific nucleic acid inhibitor depend on its structure and mechanism of action as the 3D shape of Apts is typically more sensitive to heat than siRNA duplexes or miRNA stem-loops structures. While those considerations may not be critical for processes during which the nucleic acids are only mixed with the preformed nanoparticles (PEI polymer, dendrimers, lipofectamine®), these parameters are central and need to be carefully monitored when nucleic acids are encapsulated within the carrier [157,158]. Another critical manufacturing issue has been the difficulty of scaling-up formulation methods based on lipid film hydration followed by size reduction [120].
Solely aqueous formulation of hydrophobic cationic polymers for efficient gene delivery
2021, International Journal of PharmaceuticsAdvanced polymers for nonviral gene delivery
2019, Nucleic Acid Nanotheranostics: Biomedical ApplicationsRecent advances of PLGA micro/nanoparticles for the delivery of biomacromolecular therapeutics
2018, Materials Science and Engineering CCitation Excerpt :On the other hand, direct adsorption/condensation of DNA or RNA molecules into PLGA MPs/NPs is rather difficult due to the negative charges of both entities. Similarly to the loading of proteins, DNA has been encapsulated into PLGA NPs by the dispersion in the first aqueous phase of w/o/w emulsion [64–66]. However, due to the labile nature of nucleic acids as a major limiting factor, conditions used in the NPs preparation, such as sonication, may create shear stresses, leading to their degradation [64,67].
Repurposing itraconazole to the benefit of skin cancer treatment: A combined azole-DDAB nanoencapsulation strategy
2018, Colloids and Surfaces B: Biointerfaces