Cross-Linked Enzyme Aggregates as Versatile Tool for Enzyme Delivery: Application to Polymeric Nanoparticles

Area of Science:

• Nanomedicine
• Biotechnology
• Enzyme Engineering

Background:

• Polymeric nanoparticles (NPs) are promising for drug delivery, but loading therapeutic enzymes efficiently remains a challenge.
• Enzyme replacement therapy (ERT) is crucial for treating genetic disorders caused by enzyme deficiencies.

Purpose of the Study:

• To develop a novel nanocarrier system for efficient delivery of therapeutic enzymes.
• To evaluate the encapsulation efficiency, activity retention, and in vitro efficacy of enzyme-loaded NPs for ERT.

Main Methods:

• Synthesis of poly(lactide-co-glycolide) (PLGA) NPs encapsulating cross-linked enzyme aggregates (CLEAs).
• Encapsulation and activity assessment of four key enzymes: PPT1, GALC, aGLU, and bGLU.
• In vitro delivery and functional recovery studies using cultured cells and primary fibroblasts from NCL1 disease patients.

Main Results:

• The CLEA encapsulation method achieved excellent enzyme activity retention (around 60%).
• Functional nanostructures were created, demonstrating targeted delivery and efficient release of enzymes.
• PPT1 CLEA NPs successfully restored enzymatic activity in NCL1 fibroblasts.

Conclusions:

• The developed CLEA NP system offers a viable strategy for delivering active therapeutic enzymes.
• This nanomedicine approach shows potential for improving enzyme replacement therapies for genetic diseases.
• The system enables functional recovery in patient-derived cells, highlighting its therapeutic promise.