Peptide/Lipid-Associated Nucleic Acids (PLANAs) as a Multicomponent siRNA Delivery System

Area of Science:

• Biotechnology
• Nanomedicine
• Molecular Biology

Background:

• RNA interference (RNAi) using small interfering RNA (siRNA) offers potential for cancer therapy by inhibiting protein overexpression.
• The clinical application of siRNA is hindered by challenges in cellular delivery, including siRNA's negative charge and instability.
• Developing effective delivery systems is crucial for harnessing the therapeutic potential of RNAi.

Purpose of the Study:

• To design and characterize a novel multicomponent nanoparticle system for enhanced siRNA delivery.
• To evaluate the efficacy of these nanoparticles in encapsulating, releasing, and delivering siRNA into cancer cells.
• To assess the protein silencing capabilities of the developed system in a triple-negative breast cancer model.

Main Methods:

• Creation of peptide lipid-associated nucleic acids (PLANAs) using peptides and lipids (DOTAP, DOPE, cholesterol, phosphatidylcholine).
• Formulation of nanoparticles (~100 nm) via mini-extrusion, with evaluation of peptide incorporation, siRNA encapsulation efficiency (>95%), release profiles, and toxicity.
• Assessment of cellular uptake and protein silencing of model proteins (Src, RPS6KA5) in MDA-MB-231 cells.

Main Results:

• PLANAs demonstrated high siRNA encapsulation efficiency (>95%) and controlled release.
• Formulations without positively charged lipids showed negligible toxicity.
• PLANAs C and D achieved optimal cellular uptake (80-90% siRNA-positive cells), with PLANA D effectively silencing Src and RPS6KA5 expression in breast cancer cells.

Conclusions:

• Peptide lipid-associated nucleic acids (PLANAs) represent a promising multicomponent system for safe and efficient siRNA delivery.
• The developed PLANAs exhibit significant potential for targeted gene silencing in cancer therapy.
• Further in vivo investigation is warranted to explore the therapeutic applications of PLANAs.