Potential Biomarkers of Acute Ischemic Stroke Etiology Revealed by Mass Spectrometry-Based Proteomic Characterization of Formalin-Fixed Paraffin-Embedded Blood Clots

Rosanna Rossi; Oana Madalina Mereuta; Mariel Barbachan E Silva; Sara Molina Gil; Andrew Douglas; Abhay Pandit; Michael Gilvarry; Ray McCarthy; Shane O'Connell; Ciara Tierney; Klearchos Psychogios; Georgios Tsivgoulis; István Szikora; Turgut Tatlisumak; Alexandros Rentzos; John Thornton; Pilib Ó Broin; Karen M Doyle

doi:10.3389/fneur.2022.854846

Potential Biomarkers of Acute Ischemic Stroke Etiology Revealed by Mass Spectrometry-Based Proteomic Characterization of Formalin-Fixed Paraffin-Embedded Blood Clots

Front Neurol. 2022 Apr 19:13:854846. doi: 10.3389/fneur.2022.854846. eCollection 2022.

Authors

Rosanna Rossi^{1

2}, Oana Madalina Mereuta^{1

2}, Mariel Barbachan E Silva³, Sara Molina Gil^{1

2}, Andrew Douglas^{1

2}, Abhay Pandit², Michael Gilvarry⁴, Ray McCarthy⁴, Shane O'Connell³, Ciara Tierney^{1

2}, Klearchos Psychogios⁵, Georgios Tsivgoulis⁶, István Szikora⁷, Turgut Tatlisumak^{8

9}, Alexandros Rentzos¹⁰, John Thornton¹¹, Pilib Ó Broin³, Karen M Doyle^{1

2}

Affiliations

¹ Department of Physiology and Galway Neuroscience Centre, School of Medicine, National University of Ireland, Galway, Ireland.
² CÚRAM-SFI Research Centre for Medical Devices, National University of Ireland Galway, Galway, Ireland.
³ School of Mathematical and Statistical Sciences, National University of Ireland Galway, Galway, Ireland.
⁴ Cerenovus, Galway, Ireland.
⁵ Metropolitan Hospital, Stroke Unit, Piraeus, Greece.
⁶ Second Department of Neurology, National and Kapodistrian University of Athens, "Attikon" University Hospital, Athens, Greece.
⁷ Department of Neurointerventions, National Institute of Clinical Neurosciences, Budapest, Hungary.
⁸ Department of Neurology, Sahlgrenska University Hospital, Gothenburg, Sweden.
⁹ Department of Clinical Neuroscience, Institute of Neuroscience and Physiology, Sahlgrenska Academy at University of Gothenburg, Gothenburg, Sweden.
¹⁰ Department of Interventional and Diagnostic Neuroradiology, Sahlgrenska University Hospital, University of Gothenburg, Gothenburg, Sweden.
¹¹ Department of Radiology, Royal College of Surgeons in Ireland, Beaumont Hospital, Dublin, Ireland.

Abstract

Background and aims: Besides the crucial role in the treatment of acute ischemic stroke (AIS), mechanical thrombectomy represents a unique opportunity for researchers to study the retrieved clots, with the possibility of unveiling biological patterns linked to stroke pathophysiology and etiology. We aimed to develop a shotgun proteomic approach to study and compare the proteome of formalin-fixed paraffin-embedded (FFPE) cardioembolic and large artery atherosclerotic (LAA) clots.

Methods: We used 16 cardioembolic and 15 LAA FFPE thrombi from 31 AIS patients. The thrombus proteome was analyzed by label-free quantitative liquid chromatography-tandem mass spectrometry (LC-MS/MS). MaxQuant v1.5.2.8 and Perseus v.1.6.15.0 were used for bioinformatics analysis. Protein classes were identified using the PANTHER database and the STRING database was used to predict protein interactions.

Results: We identified 1,581 protein groups as part of the AIS thrombus proteome. Fourteen significantly differentially abundant proteins across the two etiologies were identified. Four proteins involved in the ubiquitin-proteasome pathway, blood coagulation or plasminogen activating cascade were identified as significantly abundant in LAA clots. Ten proteins involved in the ubiquitin proteasome-pathway, cytoskeletal remodeling of platelets, platelet adhesion or blood coagulation were identified as significantly abundant in cardioembolic clots.

Conclusion: Our results outlined a set of 14 proteins for a proof-of-principle characterization of cardioembolic and LAA FFPE clots, advancing the proteome profile of AIS human thrombi and understanding the pathophysiology of ischemic stroke.

Keywords: FFPE proteomics; biomarkers stroke etiology; mass spectrometry proteomics; stroke biomarkers; thrombus proteome.

Copyright © 2022 Rossi, Mereuta, Barbachan e Silva, Molina Gil, Douglas, Pandit, Gilvarry, McCarthy, O'Connell, Tierney, Psychogios, Tsivgoulis, Szikora, Tatlisumak, Rentzos, Thornton, Ó Broin and Doyle.