Antisense oligonucleotides (ASOs) are DNA-based, disease-modifying drugs. Clinical trials with 2'-O-methoxyethyl (2’MOE) ASOs have reported dose- and sequencespecific lowering of platelet counts according to two phenotypes. Phenotype 1 is a moderate (but not clinically severe) drop in platelet count. Phenotype 2 is rare, severe thrombocytopenia. This paper will focus on the underlying cause of the more common Phenotype 1, investigating the effects of ASOs on platelet production and platelet function.
Five phosphorothioate ASOs were included: three 2’MOE sequences; 487660 (no effects on platelet count), 104838 (associated with Phenotype 1), 501861 (effects unknown) and two CpG sequences; 120704 and ODN 2395 (known to activate platelets). Human cord blood-derived megakaryocytes were treated with these ASOs to study effects on proplatelet production. Platelet activation (surface P-selectin) and platelet-leukocyte aggregates (PLAs) were analyzed in ASO-treated blood from healthy human volunteers.
None of the ASOs inhibited proplatelet production from human megakaryocytes. All the ASOs were shown to bind to the platelet receptor glycoprotein VI (GPVI, KD ~0.2-1.5μM). CpG ASOs had the highest affinity to GPVI and the most potent platelet activating effects and PLA formation. 2’MOE ASO 487660 had no detectable platelet effects, while 2’MOE ASOs 104838 and 501861 triggered moderate platelet activation and SYK-dependent formation of PLAs. Donors with higher platelet GPVI levels had larger ASO-induced platelet activation.
Sequence-dependent ASO-induced platelet activation and PLAs may explain Phenotype 1- moderate drops in platelet count. Platelet GPVI levels could be useful as a screening tool to identify patients at higher risk of ASO-induced platelet side effects.
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