Mangoud Nadia, Hegazy Mohamed I, Estfanous Shady, Ali Sahar A
Center of Excellence, Helwan Structure Biology Research, Cairo, Egypt; Biochemistry and Molecular Biology Department, Faculty of Pharmacy, Helwan University, Cairo, Egypt.
Neurology Department, Faculty of Medicine, Cairo University, Cairo, Egypt.
Biochim Biophys Acta Mol Basis Dis. 2025 Oct;1871(7):167922. doi: 10.1016/j.bbadis.2025.167922. Epub 2025 May 26.
Multiple sclerosis (MS) and Neuromyelitis Optica (NMO) are complex, multifactorial diseases that often present with similar clinical symptoms, leading to frequent misdiagnosis, especially in cases where NMO is aquaporin-4 seronegative. Identifying the mechanisms underlying MS may help uncover disease biomarkers and facilitate timely diagnosis, and MS treatment. There is an urgent need for differential diagnostic and prognostic tools for these devastating diseases to ensure appropriate therapy and improve patient outcomes.
This study enrolled 252 participants, divided into four groups. Group I (Relapsing-Remitting MS: RRMS group), Group II (Secondary Progressive MS: SPMS group), Group III (NMO group), and Group IV (Healthy controls), with matched age and sex. Blood samples were collected from all participants for determination of serum NRG1. Total RNA was also extracted from the whole blood then utilized for cDNA synthesis, qPCR was performed and the expression levels of lncRNA 74.1 and miR-324-3p were calculated.
NRG1 levels were significantly lower in all patient groups (RRMS, SPMS, NMO) compared to healthy controls (p-values <0.05), with the most pronounced reduction observed in NMO. ROC curve analysis indicated that NRG1 could distinguish NMO from MS with an accuracy (AUC) of 0.70. miR-324-3p was notably upregulated in RRMS and NMO compared to controls and showed strong potential in differentiating NMO from other conditions. A significant difference in miR-324-3p was observed between RRMS and SPMS (p = 0.0012). In contrast, lncRNA 74.1 did not exhibit significant diagnostic or prognostic value across the groups. Correlation analyses revealed no significant relationships between NRG1 and miR-324-3p or lncRNA 74.1 in any patient group, though a significant positive correlation was found between lncRNA 74.1 and miR-324-3p in NMO and healthy controls.
In this study, we reaffirm the previously observed reduction in NRG1 levels in NMO patients but extend these findings by elucidating a novel regulatory axis involving lncRNA 74.1 and miR-324-3p. Our results highlight NRG1 as a valuable biomarker with strong potential to improve diagnostic differentiation between MS and NMO, particularly in cases where conventional diagnostics are inconclusive. Moreover, we identify miR-324-3p as a promising marker for distinguishing RRMS from other phenotypes, with possible prognostic value in predicting transition to SPMS. Together, these findings not only support the diagnostic relevance of NRG1 but also introduce new mechanistic insights that may enhance both diagnostic precision and future therapeutic strategies.
多发性硬化症（MS）和视神经脊髓炎（NMO）是复杂的多因素疾病，常表现出相似的临床症状，导致误诊频繁，尤其是在NMO水通道蛋白4血清阴性的情况下。确定MS的潜在机制可能有助于发现疾病生物标志物，促进及时诊断和MS治疗。迫切需要针对这些破坏性疾病的鉴别诊断和预后工具，以确保适当治疗并改善患者预后。
本研究招募了252名参与者，分为四组。第一组（复发缓解型MS：RRMS组），第二组（继发进展型MS：SPMS组），第三组（NMO组）和第四组（健康对照），年龄和性别匹配。采集所有参与者的血样以测定血清NRG1。还从全血中提取总RNA，然后用于cDNA合成，进行qPCR并计算lncRNA 74.1和miR-324-3p的表达水平。
与健康对照相比，所有患者组（RRMS、SPMS、NMO）的NRG1水平均显著降低（p值<0.05），其中NMO组降低最为明显。ROC曲线分析表明，NRG1能够以0.70的准确度（AUC）区分NMO和MS。与对照组相比，miR-324-3p在RRMS和NMO中显著上调，在区分NMO与其他疾病方面显示出强大潜力。RRMS和SPMS之间观察到miR-324-3p存在显著差异（p = 0.0012）。相比之下，lncRNA 74.1在各组中未表现出显著的诊断或预后价值。相关性分析显示，在任何患者组中，NRG1与miR-324-3p或lncRNA 74.1之间均无显著关系，尽管在NMO和健康对照中lncRNA 74.1与miR-324-3p之间存在显著正相关。
在本研究中，我们再次证实了之前观察到的NMO患者NRG1水平降低的现象，但通过阐明涉及lncRNA 74.1和miR-324-3p的新调控轴扩展了这些发现。我们的结果突出了NRG1作为一种有价值的生物标志物，在改善MS和NMO的诊断鉴别方面具有强大潜力，特别是在传统诊断不确定的情况下。此外，我们确定miR-324-3p是区分RRMS与其他表型的有前景的标志物，在预测向SPMS转变方面可能具有预后价值。总之，这些发现不仅支持了NRG1的诊断相关性，还引入了新的机制见解，可能会提高诊断准确性和未来的治疗策略。
