Rapidly Transducing and Spatially Localized Magnetofection Using Peptide-Mediated Non-Viral Gene Delivery Based on Iron Oxide Nanoparticles

Abstract

Non-viral delivery systems are generally of low efficiency, which limits their use in gene therapy and editing applications. We previously developed a technology termed glycosaminoglycan (GAG)-binding enhanced transduction (GET) to efficiently deliver a variety of cargos intracellularly; our system employs GAG-binding peptides, which promote cell targeting, and cell penetrating peptides (CPPs), which enhance endocytotic cell internalization. Herein, we describe a further modification by combining gene delivery and magnetic targeting with the GET technology. We associated GET peptides, plasmid (p)DNA, and iron oxide superparamagnetic nanoparticles (MNPs), allowing rapid and targeted GET-mediated uptake by application of static magnetic fields in NIH3T3 cells. This produced effective transfection levels (significantly higher than the control) with seconds to minutes of exposure and localized gene delivery two orders of magnitude higher in targeted over non-targeted cell monolayers using magnetic fields (in 15 min exposure delivering GFP reporter pDNA). More importantly, high cell membrane targeting by GET-DNA and MNP co-complexes and magnetic fields allowed further enhancement to endocytotic uptake, meaning that the nucleic acid cargo was rapidly internalized beyond that of GET complexes alone (GET-DNA). Magnetofection by MNPs combined with GET-mediated delivery allows magnetic field-guided local transfection in vitro and could facilitate focused gene delivery for future regenerative and disease-targeted therapies in vivo.

Magnetic Nanoparticles as a Component of Peptide-Based DNA Delivery System for Suicide Gene Therapy of Uterine Leiomyoma

“…In fact, in a study by Shalaby and co-authors, the only application of magnetic force for UL gene therapy in rats is described. The authors have utilized magnetic resonance to generate a localized field in which they injected the virus into the UL tissue and restricted its spread into nearby tissues [18]. Several works devoted to magnetofection on animal models have been published recently.…”

“…Association of MNPs with non-viral carriers allows increasing the efficiency of delivery significantly. Thus, binding of DNA-polyplexes formed with cell-penetrated peptides to magnetic nanoparticles showed high efficiency of gene delivery into the cells, while reducing the time required to achieve cell membranes [18]. Magnetofection with adenoviral delivery was successfully applied in research devoted to uterine fibroids and fibroid-associated tumor stem cells.…”

Direct contact-mediated non-viral gene therapy using thermo-sensitive hydrogel-coated dressings

“…2), the highest viscosity above the critical gel temperature was seen in PF15MC2, and this formulation is also compatible with the GET-NPs, which makes it more suitable for application in trans-tissue drug delivery. We have previously published magnetically patterned GET-mediated transfection without hydrogels [60]. This demonstration through direct contact, and not through physical targeting through magnets demonstrates that efficacious gene transfer can be achieved from simply placing the transfection directly on the target area.…”

“…5A, C; Position: IN). We positioned a detachable coverslip as the targeted area (previously developed within our group [59,60]) within a homogeneous cell monolayer and patterned Azo-wipe™ material to place GET-NP laden hydrogel over the cells positioned on the coverslip (IN area) versus non-laden hydrogel on the rest of the material (OUT area) (Fig. 5C).…”

Aspects of high-performance and bio-acceptable magnetic nanoparticles for biomedical application

“…Recently, NIH3T3 fibroblast cells have been transfected by MNPs-glycosaminoglycan (GAG)-plasmid (p)DNA trio under a static magnetic field. GAG-binding enhanced transduction (GET) is proposed to proficiently deliver a diverse cargo intracellularly as a non-viral gene delivery system [104] .

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