Down-regulated lysosomal processing improved pegylated lipopolyplex-mediated gene transfection.

Area of Science:

• Biotechnology
• Gene Therapy
• Nanomedicine

Background:

• Nonviral lipid-based gene delivery vectors offer stability and circulation benefits.
• Surface PEGylation of these vectors can significantly reduce transfection efficiency.
• Lysosomal processing is a key factor limiting gene delivery efficacy.

Purpose of the Study:

• To investigate if omeprazole can enhance gene transfection efficiency of PEGylated Lipid-Mu-DNA (LMD) vectors.
• To determine if omeprazole down-regulates lysosomal processing to improve LMD delivery.
• To assess the effect of omeprazole on LMD cellular uptake and distribution.

Main Methods:

• LMD nanoparticles were formulated using cationic liposomes, peptide Mu, and plasmid DNA.
• Cellular uptake, subcellular distribution, and transfection efficiencies were analyzed using flow cytometry, confocal microscopy, and luciferase assays.
• Experiments were conducted in multiple human and mouse cell lines and in H1299 xenograft tumor models.

Main Results:

• Omeprazole treatment increased intracellular pH of acidic organelles and altered lysosomal compartments.
• Confocal microscopy revealed increased distribution of gene materials to the cytoplasm.
• Omeprazole significantly enhanced PEGylated LMD transfection efficiency in vitro and in vivo.

Conclusions:

• Omeprazole can serve as an effective adjuvant to improve gene transfection efficiency.
• This strategy shows promise for enhancing the efficacy of PEGylated gene delivery vectors.
• Omeprazole represents a potential therapeutic aid for gene therapy applications.