Integrated Multiomics Analysis Identifies a Novel Biomarker Associated with Prognosis in Intracerebral Hemorrhage

Abstract

Existing treatments for intracerebral hemorrhage (ICH) are unable to satisfactorily prevent development of secondary brain injury after ICH and multiple pathological mechanisms are involved in the development of the injury. In this study, we aimed to identify novel genes and proteins and integrated their molecular alternations to reveal key network modules involved in ICH pathology. A total of 30 C57BL/6 male mice were used for this study. The collagenase model of ICH was employed, 3 days after ICH animals were tested neurological. After it, animals were euthanized and perihematomal brain tissues were collected for transcriptome and TMT labeling-based quantitative proteome analyses. Protein-protein interaction (PPI) network, Gene Set Enrichment Analysis (GSEA), and regularized Canonical Correlation Analysis (rCCA) were performed to integrated multiomics data. For validation of hub genes and proteins, qRT-PCR and Western blot were carried out. The candidate biomarkers were further measured by ELISA in the plasma of ICH patients and the controls. A total of 2218 differentially expressed genes (DEGs) and 353 differentially expressed proteins (DEPs) between the ICH model group and control group were identified. GSEA revealed that immune-related gene sets were prominently upregulated and significantly enriched in pathways of inflammasome complex, negative regulation of interleukin-12 production, and pyroptosis during the ICH process. The rCCA network presented two highly connective clusters which were involved in the sphingolipid catabolic process and inflammatory response. Among ten hub genes screened out by integrative analysis, significantly upregulated Itgb2, Serpina3n, and Ctss were validated in the ICH group by qRT-PCR and Western blot. Plasma levels of human SERPINA3 (homologue of murine Serpina3n) were elevated in ICH patients compared with the healthy controls (SERPINA3: 13.3 ng/mL vs. 11.2 ng/mL, p = 0.015 ). Within the ICH group, higher plasma SERPINA3 levels with a predictive threshold of 14.31 ng/mL ( sensitivity = 64.3 % ; specificity = 80.8 % ; AUC = 0.742 , 95% CI: 0.567-0.916) were highly associated with poor outcome (mRS scores 4-6). Taken together, the results of our study exhibited molecular changes related to ICH-induced brain injury by multidimensional analysis and effectively identified three biomarker candidates in a mouse ICH model, as well as pointed out that Serpina3n/SERPINA3 was a potential biomarker associated with poor functional outcome in ICH patients.

Regulation of CNS pathology by Serpina3n/SERPINA3: The knowns and the puzzles

“…44,45,88 However, increasing data have challenged this concept. Clinically, SERPINA3 was reported to be elevated in the body fluid (plasma and CSF) of individuals affected by AD, 5 MS, 7 stroke, 89 intracerebral haemorrhage (ICH), 62 brain tumours 90 and normal ageing 73 funding procurement, Fuzheng Guo, Yan Wang.…”

“…However, increasing data have challenged this concept. Clinically, SERPINA3 was reported to be elevated in the body fluid (plasma and CSF) of individuals affected by AD, 5 MS, 7 stroke, 89 intracerebral haemorrhage (ICH), 62 brain tumours 90 and normal ageing 73 and positively correlated with respective disease burden/progression or poor outcomes. These clinical observations suggest that Serpina3n/SERPINA3 represents a common mechanism and a therapeutic target for shared disease pathophysiology.…”

A comprehensive analysis of the co-pathogenesis of atrial fibrillation and atherosclerosis based on bioinformatics

“…Studies have shown that C1QB and C1QC play an important role in the pathogenesis of atherosclerosis [34,39]. ITGB2 (CD18), a cell-surface glycoprotein, belongs to the integrin family and acts as a coordinator of extra-and intracellular signaling [40]. Traditionally considered important regulators of cell survival, proliferation, adhesion, and migration [41], De cient ITGB2 could also exert a neuroprotective effect by reducing in ltrating leukocytes, which decrease in ammatory cytokines and chemokines [42].…”

SERPINA3: A novel inflammatory biomarker associated with cerebral small vessel disease burden in ischemic stroke

“… 11 Conversely, recent studies have found that SERPINA3 might be a potential biomarker of poor functional outcome in patients with ICH. 12 This dual nature of SERPINA3, potentially acting as both a protective agent in ischemic stroke and a risk factor in ICH, underlines the complexity of cerebrovascular system and the diverse roles that a single protein can play within it. In our current investigation, we observed a potential link between SERPINA3 and CSVD, where higher level of SERPINA3 was associated with greater severity of WMH, a prominent imaging hallmark of CSVD.…”

“…In this study, we selected several candidate inflammatory molecules that could potentially be associated with CSVD. Serine proteinase inhibitor A3 (SERPINA3), a member of the serine protease inhibitor superfamily, has been reported to participate in pathological inflammatory processes of numerous neurological diseases such as ischemic stroke, 11 intracerebral hemorrhage, 12 and Alzheimer's disease. 13 However, despite being a potential predisposing factor for ischemic stroke, the relationship between CSVD burden and SERPINA3 has never been investigated in patients with IS.…”