Peptide-encoding gene transfer to modulate intracellular protein-protein interactions

Area of Science:

• Biochemistry
• Molecular Biology
• Cell Biology

Background:

• Intracellular protein-protein interactions (PPIs) are challenging drug targets due to the cell membrane barrier.
• Existing small molecules and peptide delivery methods are insufficient for targeting intracellular PPIs.

Purpose of the Study:

• To develop a novel platform for intracellular peptide delivery using mRNA-based gene delivery.
• To demonstrate the platform's efficacy in inhibiting a specific intracellular PPI and its downstream effects.

Main Methods:

• Engineered an mRNA construct with a dihydrofolate reductase (DHFR) sequence and a translation skipping sequence for efficient peptide production.
• Utilized mRNA delivery to introduce peptides targeting the Drp1/Fis1 interaction into cells.
• Assessed the impact on mitochondrial dynamics, metabolism, and apoptosis.

Main Results:

• Achieved efficient translation of target peptides without unnecessary protein residues.
• Successfully inhibited the Drp1/Fis1 interaction, a key regulator of mitochondrial fission.
• Demonstrated normalization of mitochondrial dynamics and metabolism, and inhibition of apoptosis.

Conclusions:

• The developed mRNA platform enables effective intracellular delivery of peptides for targeting PPIs.
• This approach offers a promising strategy for intervening in diseases associated with mitochondrial dysfunction and apoptosis.