Association of Serum Biomarkers With Post-Thrombolytic Symptomatic Intracranial Hemorrhage in Stroke: A Comprehensive Protein Microarray Analysis From INTRECIS Study

Abstract

Background: Symptomatic intracranial hemorrhage (sICH) after intravenous thrombolysis is closely related to the poor outcome of stroke.

Aims: To determine the serum biomarkers associated with sICH based on the INTRECIS study.

Methods: Enrolled patients with sICH and without any ICH were matched by propensity score matching with the ratio of 1:1. Preset 49 biomarkers were measured by protein microarray analysis. Gene Ontology and Pathway Enrichment Analysis and protein-protein interaction network (PPI) were analyzed in the identified biomarkers.

Results: Of the consecutive 358 patients, eight patients occurred with sICH, which was assigned as an sICH group, while eight matched patients without any ICH were assigned as a Non-sICH group. A total of nine biomarkers were found significantly different between groups, among which the levels of interferon (IFN)-γ and interleukin (IL)-4 were higher, while the levels of C-reactive protein (CRP), glial cell line-derived neurotrophic factor (GDNF), insulin-like growth factor-binding protein (IGFBP)-6, lymphatic vessel endothelial hyaluronan receptor (LYVE)-1, matrix metalloprotein (MMP)-2, plasminogen activator inhibitor (PAI)-1, and platelet-derived growth factor (PDGF)-AA were lower in the sICH group compared with those in the Non-sICH group.

Conclusions: Our finding indicated that baseline serum CRP, GDNF, IFN-γ, IGFBP-6, IL-4, LYVE-1, MMP-2, PAI-1, and PDGF-AA levels were associated with post-thrombolytic sICH in stroke.

Keywords: biomarkers; intravenous thrombolysis; ischemic stroke; microarray analysis; symptomatic intracranial hemorrhage; tPA.

Figure 1

Flow diagram. rtPA, recombinant tissue plasminogen activator; sICH, symptomatic intracranial hemorrhage.

Figure 2

Results of detected biomarkers in the microarray analysis. (A) The scatter plot for detected biomarkers; the X-axis represents the average of log₂ serum levels in the symptomatic intracranial hemorrhage (sICH) group, while the Y-axis represents the average of log₂ serum levels in Non-sICH group; compared with Non-sICH group, the blue point represents biomarkers with lower serum levels in sICH group, while the red point represents biomarkers with higher serum levels. sICH, symptomatic intracranial hemorrhage. (B) The volcano plot for detected biomarkers; the X-axis represents the log₂ fold-change value, while the Y-axis represents the –log₁₀ P value; the cyan point represents the biomarkers with significant difference, while the red point represents the biomarkers without significant difference. sICH, symptomatic intracranial hemorrhage. (C) The column plot for identified biomarkers; the X-axis represents the identified biomarkers, while the Y-axis represents the average of log₂ serum levels in two groups; the deep color represents the sICH group, while the light color represents the Non-sICH group. CRP, C-reactive protein; GDNF, glial cell line-derived neurotrophic factor; IFN-γ, interferon gamma; IGFBP-6, insulin-like growth factor binding protein 6; IL-4, interleukin 4; LYVE-1, lymphatic vessel endothelial hyaluronan receptor 1; MMP-2, matrix metalloprotein 2; PAI-1, plasminogen activator inhibitor 1; PDGF-AA, platelet-derived growth factor AA; sICH, symptomatic intracranial hemorrhage. (D) The heatmap for identified biomarkers; the red color represents biomarkers with higher serum levels, while the blue color represents the biomarkers with lower serum levels; the darker the color, the more significant the difference of biomarkers. CRP, C-reactive protein; GDNF, glial cell line-derived neurotrophic factor; IFN-γ, interferon gamma; IGFBP-6, insulin-like growth factor binding protein 6; IL-4, interleukin 4; LYVE-1, lymphatic vessel endothelial hyaluronan receptor 1; MMP-2, matrix metalloprotein 2; PAI-1, plasminogen activator inhibitor 1; PDGF-AA, platelet-derived growth factor AA; and sICH, symptomatic intracranial hemorrhage.

Figure 3

Protein function analysis of identified biomarkers. (A) Top 20 significantly enriched biological process of identified biomarkers; the X-axis represents the enrichment, while the Y-axis represents the biological process. (B) The molecular function enriched by identified biomarkers; the X-axis represents the enrichment, while the Y-axis represents the molecular function. (C) The cellular component enriched by identified biomarkers; the X-axis represents the enrichment, while the Y-axis represents the cellular component. (D) The pathway enriched by identified biomarkers; the X-axis represents the enrichment, while the Y-axis represents the pathway. The deeper the color, the larger the P value; the larger the circle, the bigger the counts.

Figure 4

Protein-protein interaction network of identified biomarkers. The CRP indicates C-reactive protein; GDNF, glial cell line-derived neurotrophic factor; IFNG, interferon gamma; IGFBP6, insulin-like growth factor binding protein 6; IL4, interleukin 4; LYVE1, lymphatic vessel endothelial hyaluronan receptor 1; MMP2, matrix metalloprotein 2; SERPINE1, plasminogen activator inhibitor 1; PDGFA, and platelet-derived growth factor AA.