RNase H-sensitive multifunctional ASO-based constructs as promising tools for the treatment of multifactorial complex...

Area of Science:

• Molecular Biology
• Drug Delivery Systems
• Oncology

Background:

• Combined therapies are crucial for treating complex diseases like cancer, but current drug combinations often cause toxicity.
• Targeted therapies can be circumvented by compensatory pathway activation, necessitating innovative treatment strategies.

Purpose of the Study:

• To design and evaluate a novel class of RNase H-sensitive constructs (3ASO) for the simultaneous intracellular release of multiple therapeutic oligonucleotides (ONs).
• To investigate the mechanism of recognition and cleavage of these constructs by RNase H1.
• To assess the efficacy and toxicity of 3ASO targeting HER2, Akt, and Hsp27 in HER2+ breast cancer cells.

Main Methods:

• Design of RNA·DNA hybrid constructs (3ASO) sensitive to RNase H.
• Utilized Escherichia coli RNase H1 to study construct recognition and cleavage mechanisms.
• Created 3ASO constructs targeting HER2, Akt, and Hsp27 mRNA in HER2+ breast cancer cells.
• Evaluated antiproliferative activity and toxicity in vitro.

Main Results:

• Demonstrated an unprecedented mode of RNase H recognition and cleavage dictated by construct topology.
• 3ASO constructs effectively delivered three distinct therapeutic oligonucleotides simultaneously.
• Exhibited low toxicity and significant antiproliferative activity in HER2+ breast cancer cells.

Conclusions:

• The developed 3ASO technology enables efficient single-dose administration of multiple ON drugs simultaneously.
• This approach holds significant potential for treating complex pathologies with multiple mRNA targets, overcoming limitations of current combination therapies.