6th Edition of Neurology World Conference 2026

Abstract

Part I Analysis of MeCP2 mRNA Expression between NMOSD Patients and healthy Controls

Background and aims. This study aimed to investigate the expression levels of methyl-CpG-binding protein 2 (MeCP2) mRNA in peripheral blood mononuclear cells (PBMCs) of patients with neuromyelitis optica spectrum disorder (NMOSD) and healthy controls (HCs) and examine its correlation with clinical severity.

Methods. A total of 104 Northern Chinese Han adults, including 53 NMOSD patients and 51 HCs, were included in the study. MeCP2 mRNA expression was measured.

Results and Discussion. MeCP2 mRNA expression is significantly lower in the NMOSD group compared to the HC group. A negative correlation is observed between MeCP2 mRNA expression and the Expanded Disability Status Scale (EDSS) scores, indicating that lower MeCP2 expression is associated with higher disease severity. Logistic regression analysis revealed that MeCP2 mRNA expression is an independent protective factor against the severity of acute NMOSD. MeCP2 showed lower protein expression levels in NMOSD patients compared to HCs. Enzyme-linked immunosorbent assay (ELISA) analysis revealed significantly higher levels of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-a) in the serum of acute NMOSD patients compared to HCs.

Conclusion. MeCP2 reduced expression could be involved in the pathogenesis of NMOSD and could serve as a potential biomarker for disease severity. The elevated levels of IL-6 and TNF-a indicate the presence of inflammation in NMOSD patients.

Part II mRNA and protein expression of MeCP2 decreased after inflammatory induction of THP-1 cells by LPS

Background and aims. Methyl-CpG-binding protein 2 (MeCP2) regulates gene expression and neuronal development. Dysregulation of MeCP2 implicated in neuroinflammatory disorders. This study aimed to investigate the effect of lipopolysaccharide (LPS)-induced inflammation on the mRNA and protein expression levels of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) of MeCP2 in THP-1 cells, a human monocytic cell line.

Methods. THP-1 cells were divided into two groups: LPS and control groups. THP-1 cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin mixture under standard conditions at 37°C and 5% CO₂. Cells were revived from frozen stocks, passaged, and grown to 80% confluence before initiating the experiments. In inflammatory induction, THP-1 cells were seeded in 6-well plates and treated with LPS (1μg/ml) with a complete medium, and the control group received a complete medium. After 48h of incubation, the cells were harvested, and both the precipitated cells and supernatant were collected for subsequent experiments.

Results and Discussion. The results revealed significantly higher concentrations of both IL-6 and TNF-α in the LPS group compared to the control group (P=0.006), indicating an inflammatory response in the THP-1 cells induced by LPS. mRNA and protein expression levels of MeCP2 showed a significant decrease in THP-1 cells after inflammatory induction by LPS compared to the control group. LPS could downregulate MeCP2 expression and affect gene expression and neuronal function.

Conclusion. The induction of inflammation in THP-1 cells via LPS treatment increased the expression of IL-6 and TNF-α by a decrease in both the mRNA and protein expression levels of MeCP2. MeCP2 dysregulation is potent in response to inflammation induced by LPS in THP-1 cells. MeCP2 contributes to the development of novel therapeutic strategies that target neuroinflammation-associated conditions.

Part III MeCP2 knockdown in THP-1 cells reveals elevated inflammatory factors

Background and aims. This study aimed to investigate the effects of methyl-CpG binding protein 2 (MeCP2) gene knockdown on the expression of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) in THP-1 cells.

Methods. Lentiviral transduction introduced small interfering RNAs (siRNAs) targeting the MeCP2 genes into THP-1 cells. Four types of lentiviral transduction were used, including one control virus (si-scramble) and three viruses containing different siRNAs (si-MeCP2-1, si-MeCP2-2, si-MeCP2-3). MeCP2 knockdown efficiency was evaluated by RNA extraction on THP-1 cell lines from both control and transfected groups. MeCP2 mRNA was validated using RT-qPCR. After MeCP2 knockdown, TNF-α and IL-6 were assessed in the culture supernatants of the control, si-scramble, and three si-MeCP2 groups using enzyme-linked immunosorbent assay (ELISA).

Results and Discussion. MeCP2 genes showed successful transfection of THP-1 cells by expression of inflammatory factors. MeCP2 mRNA showed no significant change in MeCP2 levels in the si-scramble group compared to the control group. si-MeCP2-2 and si-MeCP2-3 groups showed significant downregulation of MeCP2 levels. MeCP2 levels in the si-MeCP2-1 group decreased, although not statistically significant. MeCP2 TNF-α and IL-6 showed no significant change in MeCP2 levels in the si-scramble group compared to the control group. si-MeCP2-1, si-MeCP2-2, and si-MeCP2-3 groups showed significant increases in both TNF-α and IL-6 levels compared to the control group. MeCP2 knockdown increased the secretion of TNF-α and IL-6.

Conclusion. MeCP2 knockdown increased expression of proinflammatory cytokines, TNF-α, and IL-6 in THP-1 cells. MeCP2 could be a potential therapeutic target for immune-related disorders.