Double-coated PLGA nanoparticles with hierarchical surface architecture for CD44-targeted siRNA delivery

Area of Science:

• Biomaterials Science
• Nanotechnology
• RNA Therapeutics

Background:

• Efficient delivery of small interfering RNA (siRNA) is crucial for RNA therapeutics but remains a materials challenge.
• Poly(lactic-co-glycolic acid) (PLGA) nanoparticles offer biocompatibility but require interfacial engineering for effective siRNA delivery.
• Existing PLGA systems face limitations in cargo stabilization, cellular interaction, and cytosolic delivery.

Purpose of the Study:

• To develop and optimize a modular, double-coated PLGA nanoparticle (dcNP) platform for enhanced siRNA delivery.
• To engineer a nanocarrier system that integrates extracellular targeting with intracellular delivery requirements.
• To establish a scalable manufacturing process for advanced siRNA delivery vehicles.

Main Methods:

• Fabrication of a double-coated PLGA nanoparticle (dcNP) platform with a polyethyleneimine (PEI) interlayer for siRNA complexation and a hyaluronic acid (HA) outer layer for targeting.
• Optimization of the fabrication process using microfluidic technology for high yield, reproducibility, and controlled siRNA loading.
• Characterization of the dcNPs using X-ray photoelectron spectroscopy and assessment of physicochemical stability under various storage and media conditions.
• Evaluation of cellular uptake via CD44-mediated endocytosis and assessment of gene silencing efficacy in 2D and 3D cell cultures.

Main Results:

• The developed dcNPs (dcNPs2.0) demonstrated successful hierarchical multilayer assembly and robust physicochemical stability, including after lyophilization.
• Microfluidic fabrication yielded high-efficiency production with narrow size distributions and increased siRNA loading capacity.
• The HA outer layer facilitated CD44-mediated cellular uptake, leading to efficient siRNA delivery and gene silencing in MDA-MB-231 cells.
• The platform showed effective siRNA delivery and gene silencing in both 2D monolayers and 3D spheroids.

Conclusions:

• The modular dcNP platform represents a scalable and rationally engineered solution for siRNA delivery.
• This nanoplatform successfully integrates extracellular targeting (via HA) with essential intracellular delivery functions.
• The optimized dcNPs show significant potential for advancing siRNA-based therapeutic applications.