Department of Neurosurgery, Qilu Hospital, Cheeloo College of Medicine and Institute of Brain and Brain-Inspired Science, Shandong University, Jinan, 250012, China.
Shandong Key Laboratory of Brain Function Remodeling, Jinan, China.
J Neuroinflammation. 2024 Sep 15;21(1):225. doi: 10.1186/s12974-024-03226-0.
Intracranial aneurysm (IA) is a severe cerebrovascular disease, and effective gene therapy and drug interventions for its treatment are still lacking. Oxidative stress (OS) is closely associated with the IA, but the key regulatory genes involved are still unclear. Through multiomics analysis and experimental validation, we identified two diagnostic markers for IA associated with OS.
In this study, we first analyzed the IA dataset GSE75436 and conducted a joint analysis of oxidative stress-related genes (ORGs). Differential analysis, functional enrichment analysis, immune infiltration, WGCNA, PPI, LASSO, and other methods were used to identify IA diagnostic markers related to OS. Next, the functions of TLR4 and ALOX5 expression in IA and their potential targeted therapeutic drugs were analyzed. We also performed single-cell sequencing of patient IA and control (superficial temporal artery, STA) tissues. 23,342 cells were captured from 2 IA and 3 STA samples obtained from our center. Cell clustering and annotation were conducted using R software to observe the distribution of TLR4 and ALOX5 expression in IAs. Finally, the expression of TLR4 and ALOX5 were validated in IA patients and in an elastase-induced mouse IA model using experiments such as WB and immunofluorescence.
Through bioinformatics analysis, we identified 16 key ORGs associated with IA pathogenesis. Further screening revealed that ALOX5 and TLR4 were highly expressed to activate a series of inflammatory responses and reduce the production of myocytes. Methotrexate (MTX) may be a potential targeted drug. Single-cell analysis revealed a notable increase in immune cells in the IA group, with ALOX5 and TLR4 primarily localized to monocytes/macrophages. Validation through patient samples and mouse models confirmed high expression of ALOX5 and TLR4 in IAs.
Bioinformatics analysis indicated that ALOX5 and TLR4 are the most significant ORGs associated with the pathogenesis of IA. Single-cell sequencing and experiments revealed that the high expression of ALOX5 and TLR4 are closely related to IA. These two genes are promising new targets for IA therapy.
颅内动脉瘤（IA）是一种严重的脑血管疾病，目前仍缺乏有效的基因治疗和药物干预手段。氧化应激（OS）与 IA 密切相关，但涉及的关键调节基因仍不清楚。通过多组学分析和实验验证，我们确定了两个与 OS 相关的 IA 诊断标志物。
本研究首先分析了 IA 数据集 GSE75436，并对氧化应激相关基因（ORGs）进行了联合分析。采用差异分析、功能富集分析、免疫浸润、WGCNA、PPI、LASSO 等方法，鉴定与 OS 相关的 IA 诊断标志物。接着，分析了 TLR4 和 ALOX5 在 IA 中的表达及其潜在的靶向治疗药物。我们还对患者 IA 和对照（颞浅动脉，STA）组织进行了单细胞测序。从我们中心获得的 2 个 IA 和 3 个 STA 样本中捕获了 23342 个细胞。使用 R 软件对细胞聚类和注释进行分析，观察 TLR4 和 ALOX5 在 IA 中的表达分布。最后，通过 WB 和免疫荧光等实验，在 IA 患者和弹性蛋白酶诱导的 IA 小鼠模型中验证了 TLR4 和 ALOX5 的表达。
通过生物信息学分析，我们确定了 16 个与 IA 发病机制相关的关键 ORGs。进一步筛选发现，ALOX5 和 TLR4 表达上调，激活一系列炎症反应，减少肌细胞生成。甲氨蝶呤（MTX）可能是一种潜在的靶向药物。单细胞分析显示，IA 组免疫细胞明显增加，ALOX5 和 TLR4 主要定位于单核细胞/巨噬细胞。通过患者样本和小鼠模型的验证，证实了 ALOX5 和 TLR4 在 IA 中的高表达。
生物信息学分析表明，ALOX5 和 TLR4 是与 IA 发病机制最密切相关的关键 ORGs。单细胞测序和实验表明，ALOX5 和 TLR4 的高表达与 IA 密切相关。这两个基因是 IA 治疗的有前途的新靶点。
