Phosphorothioate backbone modifications of nucleotide-based drugs are potent platelet activators
Open accessAbstract
Nucleotide-based drug candidates such as antisense oligonucleotides, aptamers, immunoreceptor-activating nucleotides, or (anti)microRNAs hold great therapeutic promise for many human diseases. Phosphorothioate (PS) backbone modification of nucleotide-based drugs is common practice to protect these promising drug candidates from rapid degradation by plasma and intracellular nucleases. Effects of the changes in physicochemical properties associated with PS modification on platelets have not been elucidated so far. Here we report the unexpected binding of PS-modified oligonucleotides to platelets eliciting strong platelet activation, signaling, reactive oxygen species generation, adhesion, spreading, aggregation, and thrombus formation in vitro and in vivo. Mechanistically, the platelet-specific receptor glycoprotein VI (GPVI) mediates these platelet-activating effects. Notably, platelets from GPVI function–deficient patients do not exhibit binding of PS-modified oligonucleotides, and platelet activation is fully abolished. Our data demonstrate a novel, unexpected, PS backbone–dependent, platelet-activating effect of nucleotide-based drug candidates mediated by GPVI. This unforeseen effect should be considered in the ongoing development programs for the broad range of upcoming and promising DNA/RNA therapeutics.
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The most advanced CNS-active ASO scaffold utilizes a mixed backbone configuration with at least 70-80% PS content, maintaining efficacy and distribution with minimal risk of toxic phenotypes.
“The most advanced CNS-active ASO scaffold utilizes a mixed backbone configuration with at least 70–80% PS content. This modification maintains efficacy and distribution with minimal risk of toxic phenotypes 13, 14 via mechanisms that are not fully understood15.”
From A divalent siRNA chemical scaffold for potent and sustained modulation of gene expression throughout the central nervous system (2019) · cites this paper as [13]
The Results and Discussion sections on PS backbone modification and therapeutic relevance report platelet activation by PS-modified oligonucleotides and warn that these effects should be assessed in clinical trials. That is contrary to the claim of minimal toxic risk. The paper does not evaluate a CNS-active scaffold with 70–80% PS content, so its findings do not establish the risk of that specific mixed backbone. (LLM, conf=0.76)
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Experimental — accuracy not yet measuredThrombocytopenia (low platelet count) has been reported as a PS-mediated rare adverse event.
“In addition, we found no measurable changes in blood chemistry or cell counts (Supplementary Fig. 20), which is particularly important because thrombocytopenia (low platelet count) has been reported as a PS-mediated rare adverse event13, 14.”
From A divalent siRNA chemical scaffold for potent and sustained modulation of gene expression throughout the central nervous system (2019) · cites this paper as [13]
The Results and Discussion section, under “Relevance for PS-modified, nucleotide-based therapeutics” (page unavailable), identifies thrombocytopenia as a reported side effect of the PS-modified drug oblimersen. It does not say the event is rare or establish that the PS modification causes thrombocytopenia. (LLM, conf=0.85)
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- Authors
- Flierl, U., Nero, T. L., Lim, B., Arthur, J. F., Yao, Y., Jung, S. M., Gitz, E., Pollitt, A. Y., Zaldivia, M. T. K., Jandrot-Perrus, M., Schäfer, A., Nieswandt, B., Andrews, R. K., Parker, M. W., Gardiner, E. E., & Peter, K.
- Affiliations
- Baker Heart and Diabetes Institute (AU)
- Monash University (AU)
- Baker IDI Heart and Diabetes Institute 1 , 2 , and 3
- University of Cambridge (GB)
- University of Birmingham (GB)
- Institut National de la Santé et de la Recherche Médicale, U1148, 75877 Paris, France 7
- Medizinische Hochschule Hannover (DE)
- Rudolf Virchow Centre for Experimental Biomedicine, D-97080 Würzburg, Germany 9
- Corresponding author
- Peter, Karlheinz
- ORCID
- Venue
- Journal of Experimental Medicine
- Publisher
- Rockefeller University Press
- Published
- 2015-02-02
- Volume
- 212
- Issue
- 2
- Pages
- 129-137
- ISSN
- 0022-1007 (ISSN-L)
- 1540-9538
- PMC
- PMC4322051
- OpenAlex
- W2154236524
- Cited by
- 114
- References
- 30
- Citations by year
- 2026: 8 · 2025: 6 · 2024: 7 · 2023: 11 · 2022: 6 · 2021: 8 · 2020: 6 · 2019: 22 · 2018: 7 · 2017: 17 · 2016: 10 · 2015: 5
- FWCI
- 10.20
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- The Journal of Experimental Medicine (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion, cc-by-nc-sa)
- PubMed Central (publishedVersion, cc-by-nc-sa)
- PubMed (publishedVersion)
- CentAUR (University of Reading) (acceptedVersion)
- Minerva Access (University of Melbourne) (submittedVersion, cc-by-nc-sa)
- ANU Open Research (Australian National University) (submittedVersion)
- Funders
- British Heart Foundation (SP/13/7/30575)
- Topics
- Health Sciences › Medicine › Hematology › Platelet Disorders and Treatments
- Health Sciences › Medicine › Surgery › Heparin-Induced Thrombocytopenia and Thrombosis
- Health Sciences › Medicine › Hematology › Blood Coagulation and Thrombosis Mechanisms
- Keywords
- platelet activation, glycoprotein VI, antisense oligonucleotides, aptamers, microRNA, reactive oxygen species, platelet aggregation, thrombus formation, platelet adhesion
- MeSH terms
- Animals, Blood Platelets, Disease Models, Animal, Humans, Male, Mesenteric Arteries, Mesenteric Vascular Occlusion, Models, Molecular, Molecular Conformation, Platelet Adhesiveness, Platelet Aggregation, Platelet Membrane Glycoproteins, Protein Binding, Pulmonary Embolism, Signal Transduction, Platelet Activation, Oligonucleotides, Antisense, Reactive Oxygen Species, RNA Interference, Mice, Phosphorothioate Oligonucleotides
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