Targeted delivery of antisense oligodeoxynucleotide and small interference RNA into lung cancer cells
1Division of Molecular Pharmaceutics, School of Pharmacy, University of North Carolina at Chapel Hill, North Carolina 27599, USA.
Abstract:
Selective gene inhibition by antisense oligodeoxynucleotide (AS-ODN) or by small interference RNA (siRNA) therapeutics promises the treatment of diseases that cannot be cured by conventional drugs. However, antisense therapy is hindered due to poor stability in physiological fluids and limited intracellular uptake. To address these problems, a ligand targeted and sterically stabilized nanoparticle formulation has been developed in our lab. Human lung cancer cells often overexpress the sigma receptor and, thus, can be targeted with a specific ligand such as anisamide. AS-ODN or siRNA against human survivin was mixed with a carrier DNA, calf thymus DNA, before complexing with protamine, a highly positively charged peptide. The resulting particles were coated with cationic liposomes consisting of DOTAP and cholesterol (1:1, molar ratio) to obtain LPD (liposome-polycation-DNA) nanoparticles. Ligand targeting and steric stabilization were then introduced by incubating preformed LPD nanoparticles with DSPE-PEG-anisamide, a PEGylated ligand lipid developed earlier in our lab, by the postinsertion method. Nontargeted nanoparticles coated with DSPE-PEG were also prepared as a control. Antisense activities of nanoparticles were determined by survivin mRNA down-regulation, survivin protein down-regulation, ability to trigger apoptosis in tumor cells, tumor cell growth inhibition, and chemosensitization of the treated tumor cells to anticancer drugs. We found that tumor cell delivery and antisense activity of PEGylated nanoparticles were sequence dependent and rely on the presence of anisamide ligand. The uptake of oligonucleotide in targeted, PEGylated nanoparticles could be competed by excess free ligand. Our results suggest that the ligand targeted and sterically stabilized nanoparticles can provide a selective delivery of AS-ODN and siRNA into lung cancer cells for therapy.
Insights
Targeted nanoparticles enhance antisense therapy for lung cancer by improving drug delivery and stability. Ligand-functionalized nanoparticles show sequence-dependent activity and selective cancer cell uptake.
Area of Science:
- Nanotechnology and Drug Delivery
- Molecular Biology and Genetics
- Oncology and Cancer Therapeutics
Background:
- Antisense oligodeoxynucleotide (AS-ODN) and small interference RNA (siRNA) offer novel therapeutic avenues for intractable diseases.
- Current limitations in antisense therapy include poor stability in physiological fluids and insufficient intracellular uptake.
- Human lung cancer cells overexpress sigma receptors, presenting a target for specific ligands like anisamide.
Purpose of the Study:
- To develop a ligand-targeted and sterically stabilized nanoparticle formulation for enhanced antisense therapy.
- To evaluate the efficacy of these nanoparticles in delivering AS-ODN or siRNA to human lung cancer cells.
- To assess the therapeutic potential of targeted nanoparticles in down-regulating survivin and inhibiting tumor growth.
Main Methods:
- Formulation of liposome-polycation-DNA (LPD) nanoparticles encapsulating AS-ODN or siRNA against survivin.
- Postinsertion of DSPE-PEG-anisamide to create ligand-targeted, sterically stabilized nanoparticles.
- Assessment of nanoparticle uptake, survivin mRNA and protein down-regulation, apoptosis induction, tumor cell growth inhibition, and chemosensitization.
Main Results:
- Targeted, PEGylated nanoparticles demonstrated sequence-dependent delivery and antisense activity.
- The presence of the anisamide ligand was crucial for selective delivery and therapeutic efficacy.
- Uptake of targeted nanoparticles in cancer cells could be inhibited by excess free ligand, confirming target specificity.
Conclusions:
- Ligand-targeted and sterically stabilized nanoparticles offer a promising strategy for selective delivery of AS-ODN and siRNA.
- This nanoparticle formulation overcomes key limitations of conventional antisense therapy, enhancing its potential for lung cancer treatment.
- The developed system facilitates targeted gene inhibition in cancer cells, paving the way for improved therapeutic outcomes.
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