Upgrading Mitochondria-Targeting Peptide-Based Nanocomplexes for Zebrafish In Vivo Compatibility Assays

Area of Science:

• Biotechnology
• Gene Therapy
• Nanotechnology

Background:

• Mitochondrial gene therapy development is hindered by the absence of effective delivery systems.
• Cell-penetrating peptides (CPPs) conjugated with mitochondrial targeting sequences (MTS) are promising carriers for mitochondrial gene delivery.

Purpose of the Study:

• To evaluate PEGylated WRAP/pDNA nanocomplexes as mitochondrial gene delivery systems.
• To compare their efficacy and safety with other nanocomplexes.
• To assess their in vivo performance in a zebrafish embryo model.

Main Methods:

• PEGylation of WRAP/pDNA nanocomplexes.
• Characterization of nanocomplex size and homogeneity.
• In vitro biocompatibility and hemocompatibility assays.
• Cellular uptake and mitochondrial targeting studies in human astrocytes and lung smooth muscle cells.
• In vivo assessment of cellular internalization and toxicity in zebrafish embryos.

Main Results:

• PEGylated WRAP/pDNA nanocomplexes exhibited homogeneous sizes (100-350 nm).
• Complexes were biocompatible and hemocompatible.
• Demonstrated preferential mitochondrial targeting and efficient gene expression (ND1 protein) in cells.
• Showed effective internalization in zebrafish embryos without adverse effects on development.

Conclusions:

• MTS-CPP complexes are stable, biocompatible nanosystems for mitochondrial gene delivery.
• These nanocomplexes show significant potential for in vivo applications in mitochondrial gene therapy.
• Further in vivo research in zebrafish is warranted to explore their therapeutic performance.