Abstract Number: Esoc2026ys185 Comprehensive Profiling of Glycosylated Small Rnas in Peripheral Blood of Ischemic Stroke Patients With Different Antiplatelet Responsiveness

Abstract Background and aims To characterize the lineage features of glycosylated small RNAs (glycoRNAs) in peripheral blood mononuclear cells (PBMCs) from patients with ischemic stroke receiving antiplatelet therapy, to compare glycosylation enrichment patterns between high on-treatment platelet reactivity (HOPR) and non-HOPR patients, and to preliminarily evaluate their associations with platelet reactivity. Methods PBMC samples were collected, and paired non-enriched (input) and glyco-enriched (IP) libraries were constructed for small RNA sequencing followed by family annotation (miRNA, tsRNA, rsRNA, Y-RNA, piRNA). Differential analyses were performed using a negative-binomial generalized linear model (DESeq2) with the design formula ~ condition + assay + condition: assay, where condition represented HOPR status and assay denoted IP versus input. The interaction term, corresponding to log₂ (HOPRIP/HOPRᵢnput) / (Non-HOPRIP/Non-HOPRᵢnput), reflected the “difference-in-difference” of enrichment responses. Results The overall length distribution exhibited a bimodal pattern at 21–23 nt and 30–33 nt, dominated by miRNAs and tRNA/Y-RNA fragments, consistent with established profiles of blood and extracellular vesicle small RNAs. Compared with the Non-HOPR group, the HOPR group showed more pronounced family- and entry-level differences at the enrichment layer (IP versus input), while baseline (input) differences were limited. Y-RNAs (particularly fragments derived from RNY4) together with several tsRNAs constituted the major glycosylation-enriched populations. Some candidate enrichment indices were independently correlated with the magnitude of AA- or ADP-induced platelet activation. PBMC glycoRNAs display selective enrichment patterns associated with HOPR status. Conflict of interest Quandan Tan and Jie Yang: nothing to disclose.