Macromolecule-Loaded Hybrid Extracellular Vesicles via Ionic Lipid-Based Post-Loading for Intracellular Delivery:...

Area of Science:

• Biotechnology
• Nanotechnology
• Drug Delivery Systems

Background:

• Extracellular vesicles (EVs) are natural nanoparticles with potential as drug delivery systems (DDS).
• Current methods for loading EVs often disrupt their structure or are inefficient.
• Targeting intracellular proteins requires efficient delivery across cell membranes and endosomal escape.

Purpose of the Study:

• To develop a non-invasive method for loading macromolecules into EVs.
• To create hybrid EVs (H-EVs) capable of delivering functional molecules intracellularly.
• To evaluate the therapeutic potential of H-EVs for neurodegenerative diseases and intracellular protein targeting.

Main Methods:

• Developed a proprietary ionic lipid base (ILB) for post-loading encapsulation into EVs.
• Produced H-EVs by encapsulating proteins and nucleic acids using the ILB method.
• Assessed H-EV integrity, cellular uptake, endosomal escape, and functional delivery in vitro and in vivo.
• Utilized SH-SY5Y neuroblastoma cells to model α-synuclein aggregation and tested anti-αSyn antibody delivery.
• Administered H-EVs intravenously to mice and tracked model antibody distribution using ex vivo imaging.

Main Results:

• The ILB method enabled reproducible encapsulation of macromolecules without disrupting EV structure.
• H-EVs retained native EV surface markers and demonstrated efficient cellular uptake.
• Encapsulated molecules successfully escaped endosomes and performed their intended functions intracellularly.
• H-EVs delivering anti-αSyn antibodies suppressed α-synuclein aggregation in neuronal cells.
• H-EVs delivered anti-phospho-AKT antibodies, inducing apoptosis in target cells.
• In vivo studies showed H-EVs successfully delivered model antibodies to the brain, including regions affected by neurodegenerative diseases.

Conclusions:

• The developed H-EVs represent a versatile platform for delivering various molecules, including antibodies, to intracellular targets.
• This non-invasive encapsulation technique preserves EV integrity and function.
• H-EVs show significant promise for antibody-based therapies targeting intracellular proteins implicated in diseases like neurodegeneration.