The nucleobase guanine at the 3’-terminus of oligonucleotide RGLS4326 drives off-target AMPAR inhibition and CNS toxicity
Year: 2025
Type: article
Abstract: Designing safe and effective oligonucleotide (ON) therapeutics requires thorough understanding of structural-activity relationship (SAR) with the intended on-target(s) as well as the unintended off-target(s). Despite encouraging pharmacodynamic activity in a Phase 1b study, development of the first-generation anti-miR-17 ON RGLS4326 for the treatment of autosomal dominant polycystic kidney disease was discontinued due to dose-limiting central nervous system (CNS)-related toxicity observed in nonclinical chronic toxicity studies. Here, we provide SAR evidence that the nucleobase guanine at the 3’-terminus of RGLS4326 drives an unexpected off-target aptamer-like direct interaction with α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR), thereby causing CNS toxicity. By replacing the 3’-terminal guanine with adenine, we discover the next-generation anti-miR-17 RGLS8429 that is devoid of off-target AMPAR interaction and CNS toxicity while preserving the potency against the on-target miR-17. Here, we show a way to avoid off-target CNS effects and, more importantly, data that support the clinical development of RGLS8429. CNS toxicity was unexpectedly observed for anti-miR-17 RGLS4326 in nonclinical studies. Here, authors identify AMPA receptor inhibition as the likely culprit. Replacement of 3’-terminus guanine to adenine leads to discovery of farabursen (RGLS8429) that is devoid of CNS toxicity.
Source: Nature Communications
Authors: Tania Valencia, Laura Y. Yen, Cindy Berman, Thomas Vincent, Scott Davis, Francesca Varrone, Jianfeng Huang, Jessica Mastroianni, Morgan Carlson, Tate Owen, Amin Kamel, Denis Drygin, Garth A. Kinberger, Shanti Pal Gangwar, Maria V. Yelshanskaya, John Ridley, Robert Kirby, Jesus Alvarez, Ronak Lakhia, Vishal Patel, Alexander I. Sobolevsky, Edmund C. Lee
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $6,990
Funders: U.S. Department of Energy, Oregon Health and Science University, National Institutes of Health, Office of Science, National Cancer Institute, National Institute of General Medical Sciences, National Institute of Diabetes and Digestive and Kidney Diseases, National Institute of Neurological Disorders and Stroke, National Institute of Arthritis and Musculoskeletal and Skin Diseases, Biological and Environmental Research, SLAC National Accelerator Laboratory, Common Fund
Awards: G1213529597, G1567773112, G1921151208, The Stanford-SLAC CryoEM Center, G4582736256, G5297967758, G6099013130, G6221506569, G686058816, G7355090052, G830600299, NCCAT: National Center for CryoEM Access and Training, G9000060489, PKD1 derepression as a potential therapy for Polycystic Kidney Disease, Modulation of Heteromeric AMPA-Subtype iGluRs by Auxiliary Subunits, Structure and function of Transient Receptor Potential Channels, Structural and functional principles of activation and regulation of the transient receptor potential channel TRPV3.
OpenAlex ID: W4416794196