title: Phosphorothioate backbone modifications of nucleotide-based drugs are potent platelet activators.
abstract: 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.
authors: Baker IDI Heart and Diabetes Institute, St. Vincent's Institute of Medical Research, and Bio21 Institute, University of Melbourne, Melbourne, Victoria 3010, Australia Department of Cardiology and Angiology, Hannover Medical School, 30625 Hannover, Germany.
authors: Ulrike
authors: U
authors: Flierl
authors: Baker IDI Heart and Diabetes Institute, St. Vincent's Institute of Medical Research, and Bio21 Institute, University of Melbourne, Melbourne, Victoria 3010, Australia Baker IDI Heart and Diabetes Institute, St. Vincent's Institute of Medical Research, and Bio21 Institute, University of Melbourne, Melbourne, Victoria 3010, Australia.
authors: Tracy L
authors: TL
authors: Nero
authors: Baker IDI Heart and Diabetes Institute, St. Vincent's Institute of Medical Research, and Bio21 Institute, University of Melbourne, Melbourne, Victoria 3010, Australia.
authors: Bock
authors: B
authors: Lim
authors: Australian Centre for Blood Diseases, Monash University, Melbourne, Victoria 3004, Australia.
authors: Jane F
authors: JF
authors: Arthur
authors: Baker IDI Heart and Diabetes Institute, St. Vincent's Institute of Medical Research, and Bio21 Institute, University of Melbourne, Melbourne, Victoria 3010, Australia.
authors: Yu
authors: Y
authors: Yao
authors: Department of Biochemistry, University of Cambridge, Cambridge CB2 1GA, England, UK.
authors: Stephanie M
authors: SM
authors: Jung
authors: Centre for Cardiovascular Sciences, University of Birmingham, Birmingham B15 2TT, England, UK.
authors: Eelo
authors: E
authors: Gitz
authors: Centre for Cardiovascular Sciences, University of Birmingham, Birmingham B15 2TT, England, UK.
authors: Alice Y
authors: AY
authors: Pollitt
authors: Baker IDI Heart and Diabetes Institute, St. Vincent's Institute of Medical Research, and Bio21 Institute, University of Melbourne, Melbourne, Victoria 3010, Australia.
authors: Maria T K
authors: MT
authors: Zaldivia
authors: Institut National de la Santé et de la Recherche Médicale, U1148, 75877 Paris, France.
authors: Martine
authors: M
authors: Jandrot-Perrus
authors: Department of Cardiology and Angiology, Hannover Medical School, 30625 Hannover, Germany.
authors: Andreas
authors: A
authors: Schäfer
authors: Rudolf Virchow Centre for Experimental Biomedicine, D-97080 Würzburg, Germany.
authors: Bernhard
authors: B
authors: Nieswandt
authors: Australian Centre for Blood Diseases, Monash University, Melbourne, Victoria 3004, Australia.
authors: Robert K
authors: RK
authors: Andrews
authors: Baker IDI Heart and Diabetes Institute, St. Vincent's Institute of Medical Research, and Bio21 Institute, University of Melbourne, Melbourne, Victoria 3010, Australia Baker IDI Heart and Diabetes Institute, St. Vincent's Institute of Medical Research, and Bio21 Institute, University of Melbourne, Melbourne, Victoria 3010, Australia.
authors: Michael W
authors: MW
authors: Parker
authors: Australian Centre for Blood Diseases, Monash University, Melbourne, Victoria 3004, Australia.
authors: Elizabeth E
authors: EE
authors: Gardiner
authors: Baker IDI Heart and Diabetes Institute, St. Vincent's Institute of Medical Research, and Bio21 Institute, University of Melbourne, Melbourne, Victoria 3010, Australia karlheinz.peter@bakeridi.edu.au.
authors: Karlheinz
authors: K
authors: Peter
mesh_terms: D000818:Animals; D001792:Blood Platelets / Q000187:drug effects* / Q000378:metabolism; D004195:Disease Models, Animal; D006801:Humans; D008297:Male; D008638:Mesenteric Arteries / Q000378:metabolism / Q000473:pathology; D008641:Mesenteric Vascular Occlusion / Q000188:drug therapy / Q000378:metabolism; D051379:Mice; D008958:Models, Molecular; D008968:Molecular Conformation; D016376:Oligonucleotides, Antisense / Q000235:genetics / Q000494:pharmacology; D054735:Phosphorothioate Oligonucleotides / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D015539:Platelet Activation / Q000187:drug effects*; D010973:Platelet Adhesiveness / Q000187:drug effects; D010974:Platelet Aggregation / Q000187:drug effects; D010980:Platelet Membrane Glycoproteins / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D011655:Pulmonary Embolism / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology; D034622:RNA Interference; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects
chemical_list: D016376:Oligonucleotides, Antisense; D054735:Phosphorothioate Oligonucleotides; D010980:Platelet Membrane Glycoproteins; D017382:Reactive Oxygen Species; C098341:platelet membrane glycoprotein VI
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