Regulating Enzyme Activity via Microaggregates Mediated by Phase Separation

Area of Science:

• Biochemistry and Molecular Biology
• Biomaterials Science
• Translational Medicine

Background:

• Liquid-solid phase separation (LSPS) is a key biomolecular process for forming aggregates that influence biochemical reactions.
• Postoperative pancreatic fistula (POPF) is a severe complication characterized by pathogenic enzyme activity in the seroperitoneum.
• Current treatments for POPF lack targeted enzyme sequestration strategies.

Purpose of the Study:

• To develop a novel enzyme-regulation strategy using LSPS for treating POPF.
• To create a D-Peptide mediated Microaggregate Degradation (DPMD) system for capturing trypsin and chymotrypsin.
• To evaluate the efficacy and safety of DPMD in preclinical POPF models.

Main Methods:

• Designed DPMD with specific peptide motifs for binding cationic and hydrophobic residues, promoting β-sheet formation and self-assembly.
• Induced LSPS to form peptide-enzyme microaggregates, sequestering target proteases from the extracellular environment.
• Assessed DPMD's efficacy in rat POPF models, measuring pancreatic fluid leakage, inflammatory markers, survival rates, and performing toxicity evaluations.

Main Results:

• DPMD successfully induced LSPS, forming microaggregates that captured target enzymes (trypsin and chymotrypsin).
• In rat POPF models, DPMD significantly reduced pancreatic fluid leakage and inflammatory markers, leading to improved survival.
• Toxicity studies indicated DPMD was well-tolerated at supra-therapeutic doses, demonstrating a favorable safety profile.

Conclusions:

• DPMD represents a significant translational advance in POPF treatment by utilizing LSPS for pathogenic enzyme sequestration.
• The DPMD system effectively isolates and clears harmful enzymes, offering a promising therapeutic strategy.
• This approach broadens the potential application of LSPS in treating various enzyme-mediated diseases.