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Combined Genetic and Chemical Capsid Modifications of Adenovirus-Based Gene Transfer Vectors for Shielding and Targeting
Published on: October 26, 2018
The development of a gene vector electrostatically assembled with a polysaccharide capsule
Tomoaki Kurosaki1, Takashi Kitahara, Shigeru Kawakami
1Department of Hospital Pharmacy, Nagasaki University Hospital, 1-7-1 Sakamoto, Nagasaki 852-8501, Japan.
Biomaterials
|May 29, 2009
Summary
Researchers developed novel gene vectors using polysaccharide capsules around DNA/polyethylenimine (PEI) complexes. Chondroitin sulfate (CS) encapsulation improved safety and maintained gene delivery efficiency, suggesting a receptor-mediated uptake mechanism.
Area of Science:
- Biotechnology
- Gene Therapy
- Materials Science
Background:
- Non-viral gene vectors, such as polyethylenimine (PEI) complexed with plasmid DNA (pDNA), are crucial for gene delivery.
- Improving the safety and efficiency of these vectors remains a significant challenge in gene therapy.
- Polysaccharides offer potential for modifying vector properties due to their biocompatibility and diverse chemical structures.
Purpose of the Study:
- To develop novel gene vectors by electrostatically encapsulating pDNA/PEI complexes with various polysaccharides.
- To evaluate the physicochemical properties, safety profiles, cellular uptake, and gene transfer efficiency of these polysaccharide-coated vectors.
- To investigate the mechanism of cellular uptake for the most promising vector formulation.
Main Methods:
- pDNA/PEI complexes were formed and subsequently encapsulated with fucoidan, lambda-carrageenan, xanthan gum, alginic acid, hyaluronic acid, or chondroitin sulfate (CS).
- Nanoparticle characterization included zeta-potential measurements, agglutination assays, and cytotoxicity assessments.
- Cellular uptake and transgene efficiency were evaluated in B16-F10 cells, with mechanistic studies involving hypothermia and competitive inhibition.
Main Results:
- pDNA/PEI complexes exhibited positive zeta-potential, high agglutination, and cytotoxicity, but showed high cellular uptake and transgene efficiency.
- Polysaccharide-encapsulated complexes displayed negative zeta-potential, reduced agglutination, and lower cytotoxicity.
- Most polysaccharide-coated vectors showed limited uptake and gene expression, except for the CS-encapsulated complex, which demonstrated comparable transgene efficiency to pDNA/PEI.
- CS-encapsulated complex uptake and gene expression were inhibited by hypothermia and CS addition, indicating CS-specific receptor-mediated endocytosis.
Conclusions:
- Electrostatically encapsulating pDNA/PEI complexes with polysaccharides can significantly reduce their cytotoxicity and agglutination.
- Chondroitin sulfate (CS) encapsulation yields a safe and effective non-viral gene vector with efficient gene transfer capabilities.
- The cellular uptake of CS-encapsulated vectors is mediated by CS-specific receptors through an energy-dependent pathway, offering a targeted gene delivery strategy.
