Gene Regulation Using Nanodiscs Modified with HIF-1-α Antisense Oligonucleotides

Area of Science:

• Biotechnology
• Nanomedicine
• Molecular Biology

Background:

• Nucleic acid delivery faces challenges like nuclease degradation, endosome entrapment, and rapid clearance.
• Lipid-based nanotechnologies offer biocompatibility but struggle with stability and cargo retention.
• Antisense oligonucleotides (ASOs) show therapeutic potential but require effective delivery systems.

Purpose of the Study:

• To develop and evaluate a synthetic nanodisc (ND) scaffold for delivering anti-HIF-1-α antisense oligonucleotides (ASOs).
• To assess the efficacy of ND-ASO conjugates in reducing HIF-1-α mRNA levels in cellular models.
• To investigate the cellular uptake mechanisms and stability of the functionalized nanodiscs.

Main Methods:

• Synthetic nanodiscs were constructed using phosphoglycerolipids and an ApoA1 mimetic peptide.
• Anti-HIF-1-α ASOs were covalently conjugated to the nanodiscs via maleimide-thiol chemistry.
• Cellular uptake, nuclease resistance, and HIF-1-α mRNA knockdown were evaluated in multiple cell lines.

Main Results:

• The DNA-nanodisc conjugates were stable, nuclease-resistant, and rapidly internalized by cells.
• Nanodisc-mediated delivery led to significant reduction of HIF-1-α mRNA levels without transfection agents.
• Cellular uptake was partially mediated by Scavenger Receptor B1, and nanodisc-ASOs showed enhanced knockdown compared to soluble ASOs.

Conclusions:

• Covalently functionalized synthetic nanodiscs represent a promising platform for ASO delivery.
• This nanodisc system overcomes limitations of traditional lipid nanoparticles for nucleic acid therapeutics.
• The enhanced delivery and efficacy suggest potential for improved therapeutic strategies targeting HIF-1-α.