阻塞性睡眠呼吸暂停综合征中铁死亡相关枢纽基因的鉴定与验证

Identification and validation of ferroptosis-related hub genes in obstructive sleep apnea syndrome.

作者信息

Liu Peijun, Zhao Dong, Pan Zhou, Tang Weihua, Chen Hao, Hu Ke

机构信息

Department of Respiratory and Critical Care Medicine, Renmin Hospital of Wuhan University, Wuhan, China.

Department of Radiology, The Central Hospital of Enshi Tujia and Miao Autonomous Prefecture, Enshi, China.

出版信息

Front Neurol. 2023 Mar 2;14:1130378. doi: 10.3389/fneur.2023.1130378. eCollection 2023.

BACKGROUND

By 2020, the prevalence of Obstructive Sleep Apnea Syndrome (OSAS) in the US has reached 26. 6-43.2% in men and 8.7-27.8% in women. OSAS promotes hypertension, diabetes, and tumor growth through unknown means. Chronic intermittent hypoxia (CIH), sleep fragmentation, and increased pleural pressure are central mechanisms of OSAS complications. CIH exacerbates ferroptosis, which is closely related to malignancies. The mechanism of ferroptosis in OSAS disease progression remains unknown.

METHODS

OSAS-related datasets (GSE135917 and GSE38792) were obtained from the GEO. Differentially expressed genes (DEGs) were screened using the R software and intersected with the ferroptosis database (FerrDb V2) to get ferroptosis-related DEGs (f-DEGs). GO, DO, KEGG, and GSEA enrichment were performed, a PPI network was constructed and hub genes were screened. The TCGA database was used to obtain the thyroid cancer (THCA) gene expression profile, and hub genes were analyzed for differential and survival analysis. The mechanism was investigated using GSEA and immune infiltration. The hub genes were validated with RT-qPCR, IHC, and other datasets. Sprague-Dawley rats were randomly separated into normoxia and CIH groups. ROS, MDA, and GSH methods were used to detect CIH-induced ferroptosis and oxidative stress.

RESULTS

GSEA revealed a statistically significant difference in ferroptosis in OSAS (FDR < 0.05). HIF1A, ATM, HSPA5, MAPK8, MAPK14, TLR4, and CREB1 were identified as hub genes among 3,144 DEGs and 74 f-DEGs. HIF1A and ATM were the only two validated genes. F-DEGs were mainly enriched in THCA. HIF1A overexpression in THCA promotes its development. HIF1A is associated with CD8 T cells and macrophages, which may affect the immunological milieu. The result found CIH increased ROS and MDA while lowering GSH indicating that it could cause ferroptosis. In OSAS patients, non-invasive ventilation did not affect HIF1A and ATM expression. Carvedilol, hydralazine, and caffeine may be important in the treatment of OSAS since they suppress HIF1A and ATM.

CONCLUSIONS

Our findings revealed that the genes HIF1A and ATM are highly expressed in OSAS, and can serve as biomarkers and targets for OSAS.

背景

到2020年，美国阻塞性睡眠呼吸暂停综合征（OSAS）的患病率在男性中已达到26.6 - 43.2%，在女性中为8.7 - 27.8%。OSAS通过未知途径促进高血压、糖尿病和肿瘤生长。慢性间歇性缺氧（CIH）、睡眠片段化和胸膜压力增加是OSAS并发症的核心机制。CIH会加剧铁死亡，而铁死亡与恶性肿瘤密切相关。OSAS疾病进展中铁死亡的机制尚不清楚。

方法

从基因表达综合数据库（GEO）获取OSAS相关数据集（GSE135917和GSE38792）。使用R软件筛选差异表达基因（DEGs），并与铁死亡数据库（FerrDb V2）进行交集分析，以获得铁死亡相关的DEGs（f - DEGs）。进行基因本体论（GO）、疾病本体（DO）、京都基因与基因组百科全书（KEGG）和基因集富集分析（GSEA），构建蛋白质 - 蛋白质相互作用（PPI）网络并筛选枢纽基因。利用癌症基因组图谱（TCGA）数据库获取甲状腺癌（THCA）基因表达谱，并对枢纽基因进行差异和生存分析。使用GSEA和免疫浸润研究其机制。通过逆转录定量聚合酶链反应（RT - qPCR）、免疫组化（IHC）和其他数据集验证枢纽基因。将Sprague - Dawley大鼠随机分为常氧组和CIH组。采用活性氧（ROS）、丙二醛（MDA）和谷胱甘肽（GSH）方法检测CIH诱导的铁死亡和氧化应激。

结果

GSEA显示OSAS中铁死亡存在统计学显著差异（错误发现率<0.05）。在3144个DEGs和74个f - DEGs中，缺氧诱导因子1α（HIF1A）、共济失调毛细血管扩张突变基因（ATM）、热休克蛋白家族A成员5（HSPA5）、丝裂原活化蛋白激酶8（MAPK8）、丝裂原活化蛋白激酶14（MAPK14）、Toll样受体4（TLR4）和环磷腺苷效应元件结合蛋白1（CREB1）被确定为枢纽基因。HIF1A和ATM是仅有的两个经验证的基因。f - DEGs主要富集于THCA。THCA中HIF1A的过表达促进其发展。HIF1A与CD8 T细胞和巨噬细胞相关，这可能会影响免疫微环境。结果发现CIH会增加ROS和MDA，同时降低GSH，表明其可导致铁死亡。在OSAS患者中，无创通气不影响HIF1A和ATM的表达。卡维地洛、肼屈嗪和咖啡因可能对OSAS的治疗很重要，因为它们可抑制HIF1A和ATM。