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Inhalable Gene Delivery System Using a Cationic RAGE-Antagonist Peptide for Gene Delivery to Inflammatory Lung Cells
Chunxian Piao1, Gyeungyun Kim1, Junkyu Ha1
1Department of Bioengineering, College of Engineering, Hanyang University, Seoul 04763, Korea.
Abstract:
Acute lung injury (ALI) is a severe lung inflammatory disease. In ALI, the receptor for advanced glycation end-products (RAGE) is overexpressed in lung epithelial cells and involved in inflammatory reactions. A previous report showed that a RAGE-antagonist peptide (RAP), from high-mobility group box-1, bound to RAGE and reduced inflammatory reactions. RAP has high levels of positive amino acids, which suggests that RAP may form a complex with plasmid DNA (pDNA) by charge interactions. Because the charge density of RAP is lower than polyethylenimine (25 kDa, PEI25k), it may be able to avoid capture by the negatively charged mucus layer more easily and deliver pDNA into RAGE-positive lung cells of ALI animals by RAGE-mediated endocytosis. To prove this hypothesis, RAP was evaluated as a delivery carrier of adiponectin plasmid (pAPN) in lipopolysaccharide (LPS)-induced ALI animal models. In vitro transfection assays showed that RAP had lower transfection efficiency than PEI25k in L2 lung epithelial cells. However, in vivo administration to ALI animal models by inhalation showed that RAP had higher gene delivery efficiency than PEI25k. Particularly, due to a higher expression of RAGE in lung cells of ALI animals, the gene delivery efficiency of RAP was higher in ALI animals than that in normal animals. Delivery of the pAPN/RAP complex had anti-inflammatory effects, reducing pro-inflammatory cytokines. Hematoxylin and eosin staining confirmed that pAPN/RAP decreased inflammation in ALI models. Therefore, the results suggest that RAP may be useful as a carrier of pDNA into the lungs for ALI gene therapy.
Insights
A novel RAGE-antagonist peptide (RAP) effectively delivers gene therapy to lungs in acute lung injury (ALI) models. This peptide carrier shows promise for treating lung inflammation by targeting RAGE-positive cells.
Area of Science:
- Biomedical Engineering
- Gene Therapy
- Pulmonary Medicine
Background:
- Acute lung injury (ALI) is a severe inflammatory lung condition characterized by overexpression of the receptor for advanced glycation end-products (RAGE).
- RAGE-antagonist peptide (RAP), derived from high-mobility group box-1, binds RAGE and reduces inflammation.
- RAP's cationic nature suggests potential for plasmid DNA (pDNA) complexation and mucus layer traversal for lung-specific delivery.
Purpose of the Study:
- To evaluate RAP as a non-viral gene delivery vector for adiponectin plasmid (pAPN) in lipopolysaccharide (LPS)-induced ALI models.
- To investigate the efficacy of RAP-mediated pDNA delivery in RAGE-overexpressing lung cells of ALI animals.
- To assess the therapeutic potential of pAPN/RAP complexes in reducing lung inflammation.
Main Methods:
- In vitro transfection assays using L2 lung epithelial cells to compare RAP with polyethylenimine (PEI25k).
- In vivo gene delivery studies in LPS-induced ALI animal models via inhalation of pAPN/RAP complexes.
- Analysis of gene expression, pro-inflammatory cytokine levels, and histological examination (Hematoxylin and eosin staining) to assess therapeutic effects.
Main Results:
- In vitro, RAP exhibited lower transfection efficiency than PEI25k in L2 cells.
- In vivo inhalation studies demonstrated superior gene delivery efficiency of RAP compared to PEI25k in ALI animal models.
- Gene delivery efficiency of RAP was significantly higher in ALI animals with elevated RAGE expression than in normal animals.
- Administration of pAPN/RAP complexes reduced pro-inflammatory cytokines and decreased lung inflammation in ALI models.
Conclusions:
- RAP serves as an effective carrier for pDNA delivery to the lungs, particularly in ALI conditions with RAGE overexpression.
- The RAGE-mediated endocytosis pathway enhances RAP's gene delivery efficiency in ALI.
- RAP-based gene therapy holds potential for treating inflammatory lung diseases like ALI.
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