Intracellular fatty acid levels differentially impact target silencing by FDA-approved siRNA drugs.

With seven approved and many more anticipated over the next decade, small interfering RNA (siRNA) therapeutics have emerged as an innovative class of nucleic acid-based drugs, offering potential treatments for both rare and common diseases. A substantial portion of the patient population affected by these diseases may also present with obesity and metabolic-associated fatty liver disease (MAFLD), conditions characterized by excessive accumulation of hepatic free fatty acids (FFAs). However, the impact of intracellular FFA levels on siRNA drug efficacy has not been fully determined. In this study, hepatic HepG2 and HepaRG cells were treated with varying concentrations of oleic and palmitic acids to simulate a microcellular environment with elevated FFA levels. Efficacy in the reduction of targets at both the mRNA and protein levels was determined for three selected Food and Drug Administration (FDA)-approved siRNA drugs, patisiran, vutrisiran, and inclisiran. Our findings demonstrate strong evidence that elevated intracellular FFA levels significantly alter the efficacy of the FDA-approved siRNA drugs, impacting both mRNA and protein target reduction and highlighting a previously underexplored factor which could impact clinical outcomes. Understanding the impact on siRNA efficacy is critical for optimizing the therapeutic potential of siRNA-based treatments for patients with FFA diseases, such as obesity and MAFLD.