Lysozyme Amyloid for Synthetic RNA Delivery

Area of Science:

• Biomaterials Science
• Nanotechnology
• Immunology

Background:

• Lysozyme-based amyloid aggregates possess favorable characteristics for RNA delivery, including stability, cationic charge, and biocompatibility.
• These aggregates can form well-defined structures, making them suitable for drug carrier applications.
• Their potential for delivering immunostimulatory synthetic RNA, such as polyinosinic-polycytidylic acid (Poly(I:C)), was investigated.

Purpose of the Study:

• To evaluate lysozyme amyloid aggregates as non-viral drug carriers for RNA delivery.
• To assess the efficacy of these aggregates in delivering Poly(I:C) and small interfering RNA (siRNA).
• To confirm that observed effects were due to successful RNA delivery and not the complex itself.

Main Methods:

• Characterization of lysozyme amyloid aggregates using particle size, zeta potential, and morphology analysis.
• Assessment of RNA encapsulation efficiency.
• In vitro studies using RAW 264.7 macrophage-like cells and A549-GFP cells to evaluate RNA uptake, immune activation, and gene silencing.

Main Results:

• Poly(I:C)-loaded aggregates enhanced RNA uptake and induced significant immune activation in macrophage-like cells without toxicity.
• GFP siRNA-loaded aggregates successfully delivered functional siRNA into A549-GFP cells, leading to a notable decrease in GFP expression.
• Confocal microscopy and flow cytometry confirmed intracellular delivery and the functional impact of the delivered siRNA.

Conclusions:

• Lysozyme amyloid aggregates show significant potential as effective non-viral vectors for RNA delivery.
• These findings suggest promising applications in immunotherapy through targeted RNA delivery.
• The study validates the use of lysozyme amyloids for delivering functional RNA payloads, including immunostimulatory and gene-silencing molecules.