Fernández-Pérez Isabel, Vallverdú-Prats Marta, Rey-Álvarez Lucía, Giralt Steinhauer Eva, Ois Angel, Cuadrado-Godia Elisa, Rodriguez-Campello Ana, Suárez-Pérez Antoni, Macias-Gómez Adrià, Soriano-Tárraga Carolina, Purroy Francisco F, Arque Gloria, Tur Silvia, Cañellas Guillem, Vives-Bauza Cristofol, Segura Tomas, Serrano-Heras Gemma, Lazcano Uxue, Jiménez-Balado Joan, Jimenez-Conde Jordi
From the Neurovascular Research Group (I.F.-P., M.V.-P., L.R.-Á., E.G.S., A.O., E.C.-G., A.R.-C., A.S.-P., A.M.-G., J.J.-B., J.J.-C.), Hospital del Mar Research Institute; Neurology Department (I.F.-P., E.G.S., A.O., E.C.-G., A.R.-C., A.S.-P., A.M.-G., J.J.-C.), Hospital Del Mar; Faculty of Medicine and Life Sciences (E.G.S., A.O., E.C.-G., A.R.-C., J.J.-C.), Universitat Pompeu Fabra, Barcelona, Spain; Department of Psychiatry (C.S.-T.), Washington University School of Medicine, St. Louis, MO; Department of Neurology (F.F.P.), Arnau de Vilanova Hospital, Lleida; Clinical Neurosciences Group (F.F.P., G.A.), Biomedical Research Institute of Lleida, University of Lleida; Neurobiology Laboratory (S.T., G.C., C.V.-B.), Neurology Department, Son Espases University Hospital, Health Research Institute of Balearic Islands; Department of Biology (G.C., C.V.-B.), Universitat de les Illes Balears, Palma de Mallorca; Department of Neurology (T.S.), and Research Unit (G.S.-H.), Albacete University Hospital Complex; and Center for Cooperative Research in Biosciences (U.L.), Basque Research and Technology Alliance, Bizkaia Technology Park, Derio, Spain.
Neurology. 2025 Jan 14;104(1):e210085. doi: 10.1212/WNL.0000000000210085. Epub 2024 Dec 9.
Post-ischemic stroke (IS) outcomes vary widely among individuals, independently of clinical factors. This variability could be related to epigenetic mechanisms that regulate biological processes involved in recovery after ischemia. While several microRNAs (miRNAs) and their target genes are implicated in the pathophysiology of IS, their role in functional outcomes remains unclear. Our aim is to identify potential miRNAs associated with the 3-month outcome in patients with IS.
A discovery study was performed in patients with acute IS assessed at Hospital del Mar of Barcelona from 2009 to 2018. Main inclusion criteria were initial NIH Stroke Scale (NIHSS) score >2, hospital admission within 24 hours of IS onset, and previous functional independence. Poor 3-month outcome was defined as a modified Rankin Scale score >2. We performed miRNA next-generation sequencing on plasma samples obtained within 24 hours and performed a differential expression analysis for 2,083 miRNAs using the DESeq2 package. A replication stage was performed using real time-PCR in another multicenter cohort, with equivalent inclusion criteria and multivariate regression models. We also performed enrichment pathways analyses in both phases.
The discovery cohort included 215 patients with IS (mean age 74.7 ± 10.2 years, 54.9% male). Age, sex, and stroke severity (measured with the 24-hour NIHSS) were associated with the 3-month outcome ( < 0.05). After adjusting for these potential confounders, we found 74 miRNAs significantly associated ( < 0.05) with the 3-month poor outcome. Pathway analysis revealed significant associations with pathways related to angiogenesis, neuronal morphogenesis, transforming growth factor β (TGF-β), endothelial development, and cognition. The replication included 191 patients (mean age 78.0 ± 6.0 years, 49.7% male). We analyzed 26 miRNAs selected from the discovery stage, and 5 miRNAs replicated their association with poor outcomes ( < 0.05, fold change >1.7): miR-376c-3p, miR-4463, miR-199a-3p, miR-584-5p, and miR-134-5p.
We identified 5 miRNAs overexpressed in patients with poor 3-month outcomes after IS, which could be involved in biological processes such as cognition, neuronal morphogenesis, and TGF-β response, suggesting a potential role in brain recovery. These findings were not evaluated in infratentorial and lacunar strokes, which limits generalizability in these particular subtypes. Further investigation is needed to explore potential applicability of these findings.
缺血性中风（IS）后的个体预后差异很大，与临床因素无关。这种变异性可能与调节缺血后恢复过程中所涉及生物过程的表观遗传机制有关。虽然几种微小RNA（miRNA）及其靶基因与IS的病理生理学有关，但其在功能预后中的作用仍不清楚。我们的目的是确定与IS患者3个月预后相关的潜在miRNA。
对2009年至2018年在巴塞罗那海洋医院评估的急性IS患者进行了一项探索性研究。主要纳入标准为初始美国国立卫生研究院卒中量表（NIHSS）评分>2、在IS发作后24小时内入院以及既往功能独立。3个月预后不良定义为改良Rankin量表评分>2。我们对在24小时内采集的血浆样本进行了miRNA下一代测序，并使用DESeq2软件包对2083种miRNA进行了差异表达分析。在另一个多中心队列中使用实时聚合酶链反应进行了验证阶段研究，纳入标准和多变量回归模型相同。我们还在两个阶段进行了富集通路分析。
探索性队列包括215例IS患者（平均年龄74.7±10.2岁，54.9%为男性）。年龄、性别和中风严重程度（用24小时NIHSS测量）与3个月预后相关（P<0.05）。在对这些潜在混杂因素进行调整后，我们发现74种miRNA与3个月预后不良显著相关（P<0.05）。通路分析显示与血管生成、神经元形态发生、转化生长因子β（TGF-β）、内皮发育和认知相关的通路存在显著关联。验证阶段纳入了191例患者（平均年龄78.0±6.0岁，49.7%为男性）。我们分析了从探索阶段选出的26种miRNA，其中5种miRNA验证了其与预后不良的关联（P<0.05，倍数变化>1.7）：miR-376c-3p、miR-4463、miR-199a-3p、miR-584-5p和miR-134-5p。
我们鉴定出5种在IS后3个月预后不良的患者中过表达的miRNA，它们可能参与认知、神经元形态发生和TGF-β反应等生物过程，提示其在脑恢复中可能发挥作用。这些发现未在幕下和腔隙性中风中进行评估，这限制了这些特定亚型的普遍性。需要进一步研究以探索这些发现的潜在适用性。
