Nanodiscoidal Nucleic Acids for Gene Regulation.

Area of Science:

• Biotechnology
• Drug Delivery
• Molecular Biology

Background:

• Therapeutic nucleic acids offer gene expression control but face challenges with serum stability and endosomal entrapment.
• Lipid scaffolds, like nanodiscs (NDs), can improve nucleic acid delivery efficiency.
• NDs are self-assembling nanostructures mimicking high-density lipoproteins (HDLs).

Purpose of the Study:

• To develop and characterize nanodiscoidal nucleic acids (NNAs) for improved therapeutic nucleic acid delivery.
• To evaluate NNA stability, cellular uptake, and therapeutic efficacy.
• To assess in vivo biodistribution and therapeutic potential of NNA-delivered antisense oligonucleotides.

Main Methods:

• Covalent modification of nanodiscs (NDs) with oligonucleotides to create NNAs.
• Assessment of nuclease resistance, cellular uptake via scavenger receptor B1, and intracellular stability using FRET.
• Testing of NNAs carrying antisense oligonucleotides (ASOs) targeting HIF-1-α mRNA in cell lines and a 3D cancer spheroid model.
• In vivo biodistribution and gene silencing studies in animal models.

Main Results:

• NNAs exhibited enhanced nuclease resistance and superior cellular uptake compared to soluble nucleic acids.
• Internalized NNAs showed increased stability, confirmed by FRET analysis.
• ASO-modified NNAs demonstrated potent downregulation of HIF-1-α mRNA in vitro and in vivo, with improved efficacy at lower doses.
• In vivo studies showed preferential localization in the liver and kidneys.

Conclusions:

• NNAs represent a promising platform for enhanced delivery of therapeutic nucleic acids.
• The NNA technology overcomes key limitations of conventional nucleic acid therapies, including stability and cellular entry.
• NNAs hold significant therapeutic potential for gene silencing applications, particularly in liver and kidney targeted therapies.