Journal: Neurotherapeutics

Article Title: The EGR1/ZFP36 axis governs glycosphingolipid metabolic reprogramming in monocyte-derived macrophages in guillain-barré syndrome

doi: 10.1016/j.neurot.2026.e00895

Figure Lengend Snippet: Analysis of differentially expressed genes in the peripheral blood immune cell profile and disease-characteristic monocyte subsets of GBS patients. (a–b) Types and proportions of immune cells in the peripheral blood of GBS patients compared to healthy controls. The proportion of CD14 + monocytes was significantly increased in GBS patients. ( c –d) Types and proportions of monocyte subsets in the peripheral blood of GBS patients compared to healthy controls. CD163 + and IL1R2 + monocyte subsets were enriched in GBS patients. (e) Volcano plot of the top 5 (IL1R2, FKBP5, CD163, SAP30, and CXCR4) upregulated and top 5 (EGR1, FOSB, KLF4, CXCL8, and CCL3) downregulated differentially expressed genes in IL1R2 + monocytes from GBS patients' peripheral blood. (f) Top 20 B P enriched by GO analysis of differentially expressed genes in IL1R2 + monocytes. (g) PPI network of differentially expressed genes in IL1R2 + monocytes ranked by node score.

Article Snippet: Lentiviral vectors for EGR1 overexpression, EGR1 knockdown (sh-EGR1), and corresponding controls were constructed by GeneChem (Shanghai, China).

Techniques:

Journal: Neurotherapeutics

Article Title: The EGR1/ZFP36 axis governs glycosphingolipid metabolic reprogramming in monocyte-derived macrophages in guillain-barré syndrome

doi: 10.1016/j.neurot.2026.e00895

Figure Lengend Snippet: Analysis of DEGs in sciatic nerve transcriptome data of EAN rats. (a) Volcano plot of DEGs during the peak period of sciatic nerve in EAN rats and the top 5 DEGs with the lowest P -value. Egr1 was a significantly downregulated gene. (b) Heatmap of the expression levels of the top 5 upregulated and downregulated genes during the peak period of sciatic nerve in EAN rats in various samples, showing consistent downregulation of Cyp2c11 and Egr1 across all EAN samples. (c) Immunoinfiltration analysis of sciatic nerve ssGSEA in EAN rats, revealing significantly elevated infiltration of monocyte lineage cells. (d) Intersection Venn diagram of DEGs in GBS peripheral blood monocytes and EAN sciatic nerve, identifying 14 common genes. (e) The correlation between 14 DEGs in the sciatic nerve of EAN rats and infiltrating immune cells demonstrates that EGR1, FOS, ZFP36, and FOSB were negatively correlated with monocyte-macrophage infiltration.

Article Snippet: Lentiviral vectors for EGR1 overexpression, EGR1 knockdown (sh-EGR1), and corresponding controls were constructed by GeneChem (Shanghai, China).

Techniques: Expressing

Journal: Neurotherapeutics

Article Title: The EGR1/ZFP36 axis governs glycosphingolipid metabolic reprogramming in monocyte-derived macrophages in guillain-barré syndrome

doi: 10.1016/j.neurot.2026.e00895

Figure Lengend Snippet: The expression levels of EGR1 and ZFP36 in GBS patients, monocyte-derived macrophages, and EAN. (a) The level of EGR1 in the cerebrospinal fluid of normal (n = 25) and GBS patients (n = 48). EGR1 was significantly downregulated in GBS patients. (b) ROC curve of diagnostic efficacy of EGR1 level in cerebrospinal fluid for GBS (AUC = 0.921). ( c –d) qRT-PCR was employed to quantify the mRNA expression levels of iNOS, CD86, Arg1, CD163, EGR1, and ZFP36 in M0, M1, and M2 monocyte-derived macrophages (n = 3). EGR1 and ZFP36 were downregulated in M1 macrophages but upregulated in M2 macrophages. (e) WB was performed to analyze the protein expression of EGR1 and ZFP36 in M0 and M1 macrophages, confirming reduced expression in M1 macrophages (n = 3). (f) Immunofluorescence was employed to assess the expression and localization of EGR1 and ZFP36 in M1 macrophages, showing cytoplasmic localization and reduced fluorescence intensity (n = 3). (g–h) The expression levels of EGR1 and ZFP36 in M1 macrophages from the peripheral blood of EAN rats were confirmed by qRT-qPCR and WB analyses, further confirming their downregulation (n = 3). Data are presented as mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs HC/M0/Con group.

Article Snippet: Lentiviral vectors for EGR1 overexpression, EGR1 knockdown (sh-EGR1), and corresponding controls were constructed by GeneChem (Shanghai, China).

Techniques: Expressing, Derivative Assay, Diagnostic Assay, Quantitative RT-PCR, Immunofluorescence, Fluorescence

Journal: Neurotherapeutics

Article Title: The EGR1/ZFP36 axis governs glycosphingolipid metabolic reprogramming in monocyte-derived macrophages in guillain-barré syndrome

doi: 10.1016/j.neurot.2026.e00895

Figure Lengend Snippet: EGR1 specifically binds to ZFP36 and activates its transcription. (a) Venn diagram of the intersection between EGR1 downstream binding targets predicted based on four datasets from the TFBS database and DEGs of IL1R2 + monocytes in GBS, identifying ZFP36 and THBS1 as common targets. (b) WB detection of ZFP36 expression in M0 macrophages after overexpression and knockdown of EGR1, showing that EGR1 positively regulates ZFP36 expression (n = 3). (c) Schematic diagram showing the binding between EGR1 and ZFP36 promoter. (d). Enrichment of EGR1 binding to ZFP36 promoter was detected in M0 macrophages by ChIP-qPCR analysis, confirming specific binding of EGR1 to the ZFP36 promoter (n = 3). (e) M0 macrophages were co-transfected with ZFP36-WT or ZFP36-MUT reporter plasmids and OE-EGR1 or OE-NC plasmids; subsequently, luciferase reporter assays were performed to evaluate EGR1-mediated regulation of the ZFP36 promoter, demonstrating that EGR1 activates ZFP36 transcription through direct promoter binding (n = 3). Data are presented as mean ± SD. ∗∗∗ p < 0.001 vs ctrl/sh-NC/PMA/PMA + OE-EGR1/OE-NC + ZFP36-WT/OE-EGR1+ZFP36-WT.

Article Snippet: Lentiviral vectors for EGR1 overexpression, EGR1 knockdown (sh-EGR1), and corresponding controls were constructed by GeneChem (Shanghai, China).

Techniques: Binding Assay, Expressing, Over Expression, Knockdown, ChIP-qPCR, Transfection, Luciferase

Journal: Neurotherapeutics

Article Title: The EGR1/ZFP36 axis governs glycosphingolipid metabolic reprogramming in monocyte-derived macrophages in guillain-barré syndrome

doi: 10.1016/j.neurot.2026.e00895

Figure Lengend Snippet: Transcriptomic and metabolomic pathway enrichment analysis in EGR1-overexpressing and macrophages. (a–b) Top 20 KEGG pathways from GSEA of the ZFP36 gene set in transcriptomic data of EGR1-overexpression/knockdown macrophages. Glycosphingolipid metabolism pathways were commonly enriched in both datasets. ( c –d) Top 20 KEGG pathways from metabolomic data of EGR1-overexpression/knockdown macrophages. Sphingolipid metabolism showed positive enrichment upon EGR1 overexpression and negative enrichment upon EGR1 knockdown.

Article Snippet: Lentiviral vectors for EGR1 overexpression, EGR1 knockdown (sh-EGR1), and corresponding controls were constructed by GeneChem (Shanghai, China).

Techniques: Metabolomic, Over Expression, Knockdown

Journal: Neurotherapeutics

Article Title: The EGR1/ZFP36 axis governs glycosphingolipid metabolic reprogramming in monocyte-derived macrophages in guillain-barré syndrome

doi: 10.1016/j.neurot.2026.e00895

Figure Lengend Snippet: Enrichment patterns of sphingolipid metabolism pathway genes and metabolites in EGR1-overexpressing/knockdown macrophages. (a) Heatmap of gene expression in the sphingolipid metabolism pathway in EGR1-overexpressing and EGR1-knockdown macrophages, showing upregulation of 17 genes including HEXA and HEXB in the flag-EGR1 group. (b) KEGG network diagram integrating changes in genes and metabolites within the sphingolipid metabolism pathway in EGR1-overexpressing macrophages, demonstrating coordinated upregulation of HEXA, HEXB, and their metabolites globotriaosylceramide and lactosylceramide.

Article Snippet: Lentiviral vectors for EGR1 overexpression, EGR1 knockdown (sh-EGR1), and corresponding controls were constructed by GeneChem (Shanghai, China).

Techniques: Knockdown, Gene Expression

Journal: Neurotherapeutics

Article Title: The EGR1/ZFP36 axis governs glycosphingolipid metabolic reprogramming in monocyte-derived macrophages in guillain-barré syndrome

doi: 10.1016/j.neurot.2026.e00895

Figure Lengend Snippet: EGR1 targets ZFP36 to regulate glycosphingolipid metabolism and macrophage phenotype. (a) qRT-PCR was conducted to detect the efficiency of OE-ZFP36 and si-ZFP36 in macrophages, confirming successful overexpression and knockdown of ZFP36 for subsequent rescue experiments (n = 3). In vitro experimental groups: OE-EGR1+si-NC, OE-EGR1+si-ZFP36, OE-EGR1+si-ZFP36+OE-ZFP36. (b–c) Expression levels of glycosphingolipid metabolism markers (HEXA, HEXB, GLA) in macrophages were determined by qRT-PCR and WB (n = 3). Overexpression of EGR1 combined with ZFP36 knockdown downregulates the expression of these markers, while subsequent overexpression of ZFP36 reverses this trend. ( d –e) The proportion of M1 macrophages (CD86 + ) and M2 macrophages (CD163 + ) was determined by flow cytometry, with representative gating plots and quantitative bar charts shown, demonstrating that ZFP36 mediates EGR1 regulation of M1/M2 balance (n = 3). Data are presented as mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs OE-NC/si-NC/OE + EGR1+si-NC/OE + EGR1+si-ZFP36.

Article Snippet: Lentiviral vectors for EGR1 overexpression, EGR1 knockdown (sh-EGR1), and corresponding controls were constructed by GeneChem (Shanghai, China).

Techniques: Quantitative RT-PCR, Over Expression, Knockdown, In Vitro, Expressing, Flow Cytometry

Journal: Neurotherapeutics

Article Title: The EGR1/ZFP36 axis governs glycosphingolipid metabolic reprogramming in monocyte-derived macrophages in guillain-barré syndrome

doi: 10.1016/j.neurot.2026.e00895

Figure Lengend Snippet: Validation of EGR1 targeting ZFP36 to regulate myelination in EAN rat models. (a–b) Knockdown efficiency of ZFP36 as verified by qRT-PCR and WB, confirming successful ZFP36 knockdown in rat sciatic nerve following intrathecal siRNA injection (n = 3). (c) Assessment of hindlimb motor function using a grip strength test, showing that ZFP36 knockdown abrogated EGR1 overexpression-induced improvement in motor function (n = 6). (d) TEM was used to examine the pathological changes in myelin, revealing that ZFP36 knockdown reversed the protective effect of EGR1 on myelin structure. (e–f) qRT-PCR and WB were employed to assess the expression levels of myelin-related markers (MBP, S100B, and MPZ), demonstrating that ZFP36 mediates EGR1 regulation of myelin integrity markers (n = 3). Data are presented as mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 vs si-NC/EAN + OE-NC/EAN + OE-EGR1.

Article Snippet: Lentiviral vectors for EGR1 overexpression, EGR1 knockdown (sh-EGR1), and corresponding controls were constructed by GeneChem (Shanghai, China).

Techniques: Biomarker Discovery, Knockdown, Quantitative RT-PCR, Injection, Over Expression, Expressing