A Novel Approach for In Vitro Testing and Hazard Evaluation of Nanoformulated RyR2-Targeting siRNA Drugs Using Human...

Area of Science:

• Biotechnology
• Pharmacology
• Toxicology

Background:

• Nucleic acid (NA)-based drugs offer therapeutic promise but require rigorous safety assessments.
• Off-target effects and unintended immune stimulation are key safety concerns for NA-based therapeutics.

Purpose of the Study:

• To develop and validate an in vitro screening approach for assessing the safety of NA-based drugs.
• To identify potential toxicity pathways and immune responses induced by NA-drugs in a relevant human cell model.

Main Methods:

• Utilized human peripheral blood mononuclear cells (PBMCs) from healthy donors to model in vivo responses.
• Employed a small interfering RNA (siRNA) targeting RyR2 mRNA as a proof-of-concept NA-drug.
• Established mRNA signatures for oxidative, mitochondrial, endoplasmic reticulum stress, and autophagy pathways using known toxicants.
• Measured pro-inflammatory cytokine release to assess immune activation post-drug exposure.

Main Results:

• The developed method successfully identified specific mRNA signatures indicative of cellular stress pathways.
• Pro-inflammatory cytokine release was quantified, providing insights into potential immune stimulation.
• The approach demonstrated the ability to detect off-target effects in a relevant human cell system.

Conclusions:

• The proposed in vitro screening method is rapid, effective, and utilizes a relevant human model for evaluating NA-drug safety.
• This approach aids in identifying potential unintended adverse effects, contributing to safer drug development.
• The method can accommodate investigations into gender-specific effects and response variability.