Identification and Validation of Phagocytosis‐Regulating Factors as Potential Prognostic and Diagnostic Biomarkers for Intracerebral Hemorrhage Through Single‐Cell and Bulk Transcriptomic

ABSTRACT Intracerebral hemorrhage (ICH) is a major cause of death and disability worldwide. Despite treatment advances, reliable prognostic biomarkers are still lacking. While phagocytosis regulation is implicated in ICH pathogenesis, its potential for diagnosis and prognosis remains underexplored. Identifying phagocytic regulatory factors‐related genes (PRGs) as biomarkers could provide new insights into ICH. We used multiple datasets from the Gene Expression Omnibus (GEO) database to investigate the role of phagocytic regulatory factors in ICH. We performed Gene Set Variation Analysis (GSVA) on transcriptomic data to evaluate phagocytic regulation in ICH tissues. Single‐cell RNA sequencing (scRNA‐seq) was used to explore cell‐specific PRGs' activity. We then used machine learning for feature selection. Immunoinfiltration levels were assessed using the CIBERSORT algorithm. The biological functions of identified biomarkers were further analyzed through Gene Set Enrichment Analysis (GSEA) and a competing endogenous RNA (ceRNA) network was constructed to reveal interactions between mRNAs, miRNAs, and lncRNAs. GSVA analysis revealed significantly elevated phagocytic regulatory activity in ICH compared to normal brain tissues. Using machine learning methods, four key biomarkers—ANXA2, ANXA5, MGAT1, and VASP—were identified as potential biomarkers with diagnostic and prognostic relevance in ICH. Immunoinfiltration analysis showed a correlation between immune cell infiltration and the identified biomarkers. GSEA highlighted the involvement of these biomarkers in various biological processes. Additionally, the ceRNA network uncovered complex regulatory interactions, where miRNAs and lncRNAs modulated the expression of the identified biomarkers. The four biomarkers offer promising candidates for early detection and may also provide insights into prognosis during post‐ICH recovery. These biomarkers are linked to immune cell infiltration and phagocytic regulation, offering new insights into ICH pathogenesis and potential therapeutic targets.