Fenton Reaction-Mediated Endolysosomal Disruption for Efficient Cytosolic Protein Delivery.

Area of Science:

• Biotechnology
• Cell Biology
• Nanomedicine

Background:

• Endolysosomal entrapment hinders cytosolic protein delivery, a critical challenge in biological research and therapeutics.
• Efficient endolysosomal escape strategies are essential for advancing protein-based applications.

Purpose of the Study:

• To investigate the potential of Fenton reaction-mediated endolysosomal disruption for cytosolic protein delivery.
• To evaluate the efficacy and generality of a novel technology termed chemodynamic internalization (CDI).

Main Methods:

• Proteins and iron ions were encapsulated within calcium carbonate nanoparticles.
• Cellular uptake and intracellular distribution of proteins were analyzed post-nanoparticle administration.
• The bioactivity of delivered proteins was assessed to confirm functional integrity.

Main Results:

• Chemodynamic internalization (CDI) demonstrated efficient delivery of diverse proteins (Ribonuclease A, GFP, β-galactosidase, HRP) into the cytosol.
• Mitochondrial calcium overload enhanced the Fenton reaction, inducing lipid peroxidation and endolysosomal membrane rupture.
• Delivered proteins retained their bioactivity after cytosolic translocation.

Conclusions:

• CDI offers a robust and versatile platform for cytosolic protein delivery, overcoming endolysosomal entrapment.
• This technology has significant implications for protein-related basic science and the development of protein-based therapeutics.