Transcriptomic profiling of thrombus-derived extracellular vesicles reveals PECAM-1 as a potential inflammatory marker for cardioembolic stroke patients

Accurate etiological diagnosis ensures appropriate secondary prevention of ischemic stroke (IS) patients. However, about 25–40% of IS cases remain of undetermined origin (referred to as ESUS if an unknown embolic source is suspected), highlighting the need to improve diagnostic precision. To identif...

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Autores: Machado, Florencio J.D., Marta-Enguita, Juan, Herrera, María, Marta-Moreno, Javier, Páramo, José A., Segura, Tomás, Serrano-Heras, Gemma, Hernández-Fernández, Francisco, Arias-Salazar, Lourdes, Yélamos-Sanz, Blanca, Zandio, Beatriz, Aymerich, Nuria, Muñoz, Roberto, Bermejo, Rebeca, Roncal Mancho, Carmen, Orbe, Josune
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2025
País:España
Institución:Universidad Pública de Navarra
Repositorio:Academica-e. Repositorio Institucional de la Universidad Pública de Navarra
OAI Identifier:oai:academica-e.unavarra.es:2454/55884
Acceso en línea:https://hdl.handle.net/2454/55884
Access Level:acceso abierto
Palabra clave:Ischemic stroke
Diagnostic
Thrombus
Extracellular vesicles
Transcriptomes
And inflammation related pathways
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dc.title.none.fl_str_mv Transcriptomic profiling of thrombus-derived extracellular vesicles reveals PECAM-1 as a potential inflammatory marker for cardioembolic stroke patients
title Transcriptomic profiling of thrombus-derived extracellular vesicles reveals PECAM-1 as a potential inflammatory marker for cardioembolic stroke patients
spellingShingle Transcriptomic profiling of thrombus-derived extracellular vesicles reveals PECAM-1 as a potential inflammatory marker for cardioembolic stroke patients
Machado, Florencio J.D.
Ischemic stroke
Diagnostic
Thrombus
Extracellular vesicles
Transcriptomes
And inflammation related pathways
title_short Transcriptomic profiling of thrombus-derived extracellular vesicles reveals PECAM-1 as a potential inflammatory marker for cardioembolic stroke patients
title_full Transcriptomic profiling of thrombus-derived extracellular vesicles reveals PECAM-1 as a potential inflammatory marker for cardioembolic stroke patients
title_fullStr Transcriptomic profiling of thrombus-derived extracellular vesicles reveals PECAM-1 as a potential inflammatory marker for cardioembolic stroke patients
title_full_unstemmed Transcriptomic profiling of thrombus-derived extracellular vesicles reveals PECAM-1 as a potential inflammatory marker for cardioembolic stroke patients
title_sort Transcriptomic profiling of thrombus-derived extracellular vesicles reveals PECAM-1 as a potential inflammatory marker for cardioembolic stroke patients
dc.creator.none.fl_str_mv Machado, Florencio J.D.
Marta-Enguita, Juan
Herrera, María
Marta-Moreno, Javier
Páramo, José A.
Segura, Tomás
Serrano-Heras, Gemma
Hernández-Fernández, Francisco
Arias-Salazar, Lourdes
Yélamos-Sanz, Blanca
Zandio, Beatriz
Aymerich, Nuria
Muñoz, Roberto
Bermejo, Rebeca
Roncal Mancho, Carmen
Orbe, Josune
author Machado, Florencio J.D.
author_facet Machado, Florencio J.D.
Marta-Enguita, Juan
Herrera, María
Marta-Moreno, Javier
Páramo, José A.
Segura, Tomás
Serrano-Heras, Gemma
Hernández-Fernández, Francisco
Arias-Salazar, Lourdes
Yélamos-Sanz, Blanca
Zandio, Beatriz
Aymerich, Nuria
Muñoz, Roberto
Bermejo, Rebeca
Roncal Mancho, Carmen
Orbe, Josune
author_role author
author2 Marta-Enguita, Juan
Herrera, María
Marta-Moreno, Javier
Páramo, José A.
Segura, Tomás
Serrano-Heras, Gemma
Hernández-Fernández, Francisco
Arias-Salazar, Lourdes
Yélamos-Sanz, Blanca
Zandio, Beatriz
Aymerich, Nuria
Muñoz, Roberto
Bermejo, Rebeca
Roncal Mancho, Carmen
Orbe, Josune
author2_role author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Ciencias de la Salud
Osasun Zientziak
dc.subject.none.fl_str_mv Ischemic stroke
Diagnostic
Thrombus
Extracellular vesicles
Transcriptomes
And inflammation related pathways
topic Ischemic stroke
Diagnostic
Thrombus
Extracellular vesicles
Transcriptomes
And inflammation related pathways
description Accurate etiological diagnosis ensures appropriate secondary prevention of ischemic stroke (IS) patients. However, about 25–40% of IS cases remain of undetermined origin (referred to as ESUS if an unknown embolic source is suspected), highlighting the need to improve diagnostic precision. To identify potential biomarkers of IS etiologies, we analyzed the transcriptomic content of extracellular vesicles (EVs) from thrombi obtained after thrombectomy, including patients with cardioembolic (CE, n = 10), atherothrombotic (AT, n = 10) and ESUS (n = 10) IS (discovery cohort). mRNA levels of those differentially expressed genes in thrombus EVs were determined in microvascular brain endothelial cells after stimulation, revealing PECAM-1, as the most promising candidate. Next, PECAM-1 levels were determined in thrombi (n = 65) and serum samples (n = 95) of an independent validation cohort of IS patients, using immunohistochemistry and ELISA respectively. The transcriptome profiling and bioinformatics analysis revealed 1,514 gene transcripts in thrombus EVs, of which 114 were differentially expressed in AT vs CE etiologies. The genes upregulated in EVs from CE thrombi were enriched in pathways related to cell surface interactions at the vascular wall, leukocyte migration and neutrophil degranulation. Consistent with the increased PECAM-1 expression in thrombus EVs from CE vs AT etiologies, the endothelial expression of PECAM-1 increased after thrombin exposure and diminished upon atherogenic stimulation (oxLDL and TNFα). Moreover, thrombus expression of PECAM-1 was localized predominantly in platelet rich areas containing or surrounded by inflammatory cells and fibrin, and was higher in CE stroke thrombi. Serum levels of PECAM-1 were associated [OR (95%CI) 1.52 (1.14–2.03), p = 0.004] with the severity of stroke at Hospital admission. These findings collectively suggest that the transcriptional study of thrombus EVs may be useful to unravel the molecular pathways behind thrombus formation, and to explore local biological differences between IS etiologies. Specifically, PECAM-1, an adhesion molecule implicated in immuno-inflammatory processes, was increased in thrombi EVs and thrombi of patients with CE IS. Regarding the use of PECAM-1 as systemic biomarker, it did not discriminate IS etiologies, but was positively correlated with worse neurological outcome, suggesting a possible role of PECAM-1 in processes leading to brain injury post-IS.
publishDate 2025
dc.date.none.fl_str_mv 2025
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dc.identifier.none.fl_str_mv https://hdl.handle.net/2454/55884
url https://hdl.handle.net/2454/55884
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
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spelling Transcriptomic profiling of thrombus-derived extracellular vesicles reveals PECAM-1 as a potential inflammatory marker for cardioembolic stroke patientsMachado, Florencio J.D.Marta-Enguita, JuanHerrera, MaríaMarta-Moreno, Javier Páramo, José A.Segura, TomásSerrano-Heras, GemmaHernández-Fernández, FranciscoArias-Salazar, LourdesYélamos-Sanz, BlancaZandio, BeatrizAymerich, NuriaMuñoz, RobertoBermejo, RebecaRoncal Mancho, CarmenOrbe, JosuneIschemic strokeDiagnosticThrombusExtracellular vesiclesTranscriptomesAnd inflammation related pathwaysAccurate etiological diagnosis ensures appropriate secondary prevention of ischemic stroke (IS) patients. However, about 25–40% of IS cases remain of undetermined origin (referred to as ESUS if an unknown embolic source is suspected), highlighting the need to improve diagnostic precision. To identify potential biomarkers of IS etiologies, we analyzed the transcriptomic content of extracellular vesicles (EVs) from thrombi obtained after thrombectomy, including patients with cardioembolic (CE, n = 10), atherothrombotic (AT, n = 10) and ESUS (n = 10) IS (discovery cohort). mRNA levels of those differentially expressed genes in thrombus EVs were determined in microvascular brain endothelial cells after stimulation, revealing PECAM-1, as the most promising candidate. Next, PECAM-1 levels were determined in thrombi (n = 65) and serum samples (n = 95) of an independent validation cohort of IS patients, using immunohistochemistry and ELISA respectively. The transcriptome profiling and bioinformatics analysis revealed 1,514 gene transcripts in thrombus EVs, of which 114 were differentially expressed in AT vs CE etiologies. The genes upregulated in EVs from CE thrombi were enriched in pathways related to cell surface interactions at the vascular wall, leukocyte migration and neutrophil degranulation. Consistent with the increased PECAM-1 expression in thrombus EVs from CE vs AT etiologies, the endothelial expression of PECAM-1 increased after thrombin exposure and diminished upon atherogenic stimulation (oxLDL and TNFα). Moreover, thrombus expression of PECAM-1 was localized predominantly in platelet rich areas containing or surrounded by inflammatory cells and fibrin, and was higher in CE stroke thrombi. Serum levels of PECAM-1 were associated [OR (95%CI) 1.52 (1.14–2.03), p = 0.004] with the severity of stroke at Hospital admission. These findings collectively suggest that the transcriptional study of thrombus EVs may be useful to unravel the molecular pathways behind thrombus formation, and to explore local biological differences between IS etiologies. Specifically, PECAM-1, an adhesion molecule implicated in immuno-inflammatory processes, was increased in thrombi EVs and thrombi of patients with CE IS. Regarding the use of PECAM-1 as systemic biomarker, it did not discriminate IS etiologies, but was positively correlated with worse neurological outcome, suggesting a possible role of PECAM-1 in processes leading to brain injury post-IS.Red de Investigación Cooperativa Orientada a Resultados en Salud-Enfermedades Vasculares Cerebrales (RICORS-Ictus) RD21/0006/0008, RD24/0009/0028, and PI22/00436 financiado por el Instituto de Salud Carlos III, "Cofinanciado por la Unión Europea".BMCCiencias de la SaludOsasun Zientziak2025info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfapplication/vnd.ms-powerpointhttps://hdl.handle.net/2454/55884reponame:Academica-e. Repositorio Institucional de la Universidad Pública de Navarrainstname:Universidad Pública de NavarraInglésinfo:eu-repo/grantAgreement/ISCIII//RD21%2F0006%2F0008info:eu-repo/grantAgreement/ISCIII//RD24%2F0009%2F0028info:eu-repo/grantAgreement/ISCIII/Plan Estatal de Investigación Científica, Técnica y de Innovación 2021-2023/PI22%2F00436© The Author(s) 2025. Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material.https://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessoai:academica-e.unavarra.es:2454/558842026-06-17T12:41:47Z
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