Branched polyethylenimine-based PKCα-responsive gene carriers

Area of Science:

• Biomaterials Science
• Gene Therapy
• Nanotechnology

Background:

• Developing efficient and targeted gene delivery systems is crucial for gene therapy.
• Existing gene carriers often face challenges with endosomal escape and controlled DNA release.
• Cancer-specific activation offers a promising strategy for targeted drug delivery.

Purpose of the Study:

• To synthesize and evaluate novel branched polyethylenimine (bPEI)-based gene carriers.
• To investigate the carriers' responsiveness to cancer-specific protein kinase Cα (PKCα) activation for plasmid DNA release.
• To determine the optimal peptide content for efficient gene expression.

Main Methods:

• Synthesis of bPEI-based gene carriers via amide bond formation with peptides.
• Preparation of polyplexes with varying peptide contents and plasmid DNA.
• In vitro assessment of polyplex condensation, endosomal escape, and DNA release triggered by PKCα.
• Evaluation of gene expression levels post-transfection.

Main Results:

• The synthesized gene carriers effectively condensed plasmid DNA, forming tighter polyplexes than previous systems.
• Polyplexes demonstrated efficient endosomal escape into the cytosol after cellular uptake.
• A clear-cut release of plasmid DNA was observed in response to PKCα activation.
• The optimal peptide content for a clear-cut PKCα response was determined to be 5 mol%.

Conclusions:

• bPEI-based gene carriers functionalized with peptides offer a promising platform for targeted gene delivery.
• The carriers exhibit efficient DNA condensation, endosomal escape, and cancer-specific DNA release.
• An optimal peptide content of 5 mol% maximizes the carriers' responsiveness to PKCα for enhanced gene expression.