Protein and mRNA Delivery Enabled by Cholesteryl-Based Biodegradable Lipidoid Nanoparticles

Area of Science:

• Biomaterials Science
• Nanotechnology
• Drug Delivery Systems

Background:

• Developing safe and efficient delivery systems for therapeutic biomacromolecules (e.g., proteins, mRNA) remains a significant challenge in medicine.
• Existing delivery vectors often face limitations regarding efficacy, safety, and targeting specific tissues.
• Biodegradable and non-toxic carriers are crucial for advancing nucleic acid and protein-based therapies.

Purpose of the Study:

• To develop and characterize a novel library of cholesteryl-based, disulfide bond-containing, biodegradable cationic lipidoid nanoparticles.
• To evaluate the efficacy and safety of these novel lipidoids for in vitro and in vivo delivery of therapeutic biomacromolecules.
• To compare the performance of the newly developed lipidoids against established transfection agents.

Main Methods:

• Synthesis of a combinatorial library of cholesteryl-based, disulfide bond-containing biodegradable cationic lipidoids.
• In vitro assessment of transfection efficacy and cytotoxicity using protein and mRNA delivery models.
• In vivo studies in adult mice to evaluate biodistribution and delivery efficiency in skeletal muscle, lung, spleen, and brain following various administration routes.

Main Results:

• A subset of the developed lipidoid library demonstrated effective in vitro delivery of protein and mRNA.
• The identified lipidoids exhibited comparable transfection efficiencies to Lpf2k but with significantly lower cytotoxicity.
• Successful in vivo delivery of genome engineering proteins and mRNA was achieved in multiple organs, including muscle, lung, spleen, and brain, via different administration routes.

Conclusions:

• Cholesteryl-based, disulfide bond-containing biodegradable cationic lipidoids represent a promising new class of delivery vehicles for therapeutic biomacromolecules.
• These novel lipidoids offer a safer and effective alternative for in vitro and in vivo delivery, with potential applications in gene therapy and protein replacement.
• The demonstrated ability to deliver cargo to various tissues, including the brain, highlights the versatility of this lipidoid platform.