Abstract Background and aims Stroke is the leading cause of adult-onset disability. Although clinical factors influence stroke outcome, there is a significant variability among individuals that may be attributed to genetics and epigenetics, including DNA methylation (DNAm). We aimed to study the association between DNAm and stroke prognosis. Methods and results To that aim, we conducted a two-phase study (discovery-replication and meta-analysis) in Caucasian patients with ischemic stroke from two independent centers (BasicMar [discovery, N = 316] and St. Pau [replication, N = 92]). Functional outcome was assessed using the modified Rankin Scale (mRS) at three months after stroke, being poor outcome defined as mRS > 2. DNAm was determined using the 450K and EPIC BeadChips in whole-blood samples collected within the first 24 h. We searched for differentially methylated positions (DMPs) in 370,344 CpGs, and candidates below p -value < 10 –5 were subsequently tested in the replication cohort. We then meta-analyzed DMP results from both cohorts and used them to identify differentially methylated regions (DMRs). After doing the epigenome-wide association study, we found 29 DMPs at p -value < 10 –5 and one of them was replicated: cg24391982, annotated to thrombospondin-2 ( THBS2 ) gene ( p -value discovery = 1.54·10 –6 ; p -value replication = 9.17·10 –4 ; p -value meta-analysis = 6.39·10 –9 ). Besides, four DMRs were identified in patients with poor outcome annotated to zinc finger protein 57 homolog ( ZFP57 ), Arachidonate 12-Lipoxygenase 12S Type ( ALOX12 ), ABI Family Member 3 ( ABI3 ) and Allantoicase ( ALLC ) genes ( p -value < 1·10 –9 in all cases). Discussion Patients with poor outcome showed a DMP at THBS2 and four DMRs annotated to ZFP57, ALOX12, ABI3 and ALLC genes. This suggests an association between stroke outcome and DNAm, which may help identify new stroke recovery mechanisms.
