Endothelial progenitor cells, potential biomarkers for diagnosis and prognosis of ischemic stroke: protocol for an observational case-control study

Kamini Rakkar, Othman Othman, Nikola Sprigg, Philip Bath, Ulvi Bayraktutan, Kamini Rakkar, Othman Othman, Nikola Sprigg, Philip Bath, Ulvi Bayraktutan

Abstract

Ischemic stroke is a devastating, life altering event which can severely reduce patient quality of life. Despite years of research there have been minimal therapeutic advances. Endothelial progenitor cells (EPCs), stem cells involved in both vasculogenesis and angiogenesis, may be a potential therapeutic target. After a stroke, EPCs migrate to the site of ischemic injury to repair cerebrovascular damage, and their numbers and functional capacity may determine patients' outcome. This study aims to determine whether the number of circulating EPCs and their functional aspects may be used as biomarkers to identify the type (cortical or lacunar) and/or severity of ischemic stroke. The study will also investigate if there are any differences in these characteristics between healthy volunteers over and under 65 years of age. 100 stroke patients (50 lacunar and 50 cortical strokes) will be recruited in this prospective, observational case-controlled study. Blood samples will be taken from stroke patients at baseline (within 48 hours of stroke) and days 7, 30 and 90. EPCs will be counted with flow cytometry. The plasma levels of pro- and anti-angiogenic factors and inflammatory cytokines will also be determined. Outgrowth endothelial cells will be cultured to be used in tube formation, migration and proliferation functional assays. Primary outcome is disability or dependence on day 90 after stroke, assessed by the modified Rankin Scale. Secondary outcomes are changes in circulating EPC numbers and/or functional capacity between patient and healthy volunteers, between patient subgroups and between elderly and young healthy volunteers. Recruitment started in February 2017, 167 participants have been recruited. Recruitment will end in November 2019. West Midlands - Coventry & Warwickshire Research Ethics Committee approved this study (REC number: 16/WM/0304) on September 8, 2016. Protocol version: 2.0. The Bayraktutan Dunhill Medical Trust EPC Study was registered in ClinicalTrials.gov (NCT02980354) on November 15, 2016. This study will determine whether the number of EPCs can be used as a prognostic or diagnostic marker for ischemic strokes and is a step towards discovering if transplantation of EPCs may aid patient recovery.

Keywords: ageing; biomarkers; cortical stroke; endothelial progenitor cells; ischemic stroke; lacunar stroke; observational study; stem cells.

Conflict of interest statement

None

Figures

Figure 1
Figure 1
Experimental processing. AcLDL: Acetylated low density lipoprotein; eNOS: endothelial NO synthase; NO: nitric oxide.
Figure 2
Figure 2
Patient pathway. BI: Barthel Index; CT: computerized tomography; EPC: endothelial progenitor cell; MRI: magnetic resonance imaging; mRS: modified Rankin Scale; NIHSS: National Institutes of Health Stroke Scale; OEC: outgrowth endothelial cell.
Figure 3
Figure 3
Healthy volunteer pathway. EPC: Endothelial progenitor cell; HVs: healthy volunteers; OEC; outgrowth endothelial cell.

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