Qian Hongyan, Qian Yi, Liu Yi, Cao Jiaxin, Wang Yuhang, Yang Aihua, Zhao Wenjing, Lu Yingnan, Liu Huanxin, Zhu Weizhong
Department of Pharmacology, School of Medicine and School of Pharmacy Nantong University, Nantong, China.
Cancer Research Center Nantong, Nantong Tumor Hospital and Tumor Hospital Affiliated to Nantong University, Nantong, China.
Front Cardiovasc Med. 2023 Jan 11;9:996809. doi: 10.3389/fcvm.2022.996809. eCollection 2022.
The mechanisms of doxorubicin (DOX) cardiotoxicity were complex and controversial, with various contradictions between experimental and clinical data. Understanding the differences in the molecular mechanism between DOX-induced acute and chronic cardiotoxicity may be an ideal entry point to solve this dilemma.
Mice were injected intraperitoneally with DOX [(20 mg/kg, once) or (5 mg/kg/week, three times)] to construct acute and chronic cardiotoxicity models, respectively. Survival record and ultrasound monitored the cardiac function. The corresponding left ventricular (LV) myocardium tissues were analyzed by RNA-seq to identify differentially expressed genes (DEGs). Gene Ontology (GO), Kyoto Encyclopedia of Gene and Genome (KEGG), and Gene Set Enrichment Analysis (GSEA) found the key biological processes and signaling pathways. DOX cardiotoxicity datasets from the Gene expression omnibus (GEO) database were combined with RNA-seq to identify the common genes. Cytoscape analyzed the hub genes, which were validated by quantitative real-time PCR. ImmuCo and ImmGen databases analyzed the correlations between hub genes and immunity-relative markers in immune cells. Cibersort analyzed the immune infiltration and correlations between the hub genes and the immune cells. Logistic regression, receiver operator characteristic curve, and artificial neural network analysis evaluated the diagnosis ability of hub genes for clinical data in the GEO dataset.
The survival curves and ultrasound monitoring demonstrated that cardiotoxicity models were constructed successfully. In the acute model, 788 DEGs were enriched in the activated metabolism and the suppressed immunity-associated signaling pathways. Three hub genes (Alas1, Atp5g1, and Ptgds) were upregulated and were negatively correlated with a colony of immune-activating cells. However, in the chronic model, 281 DEGs showed that G protein-coupled receptor (GPCR)-related signaling pathways were the critical events. Three hub genes (Hsph1, Abcb1a, and Vegfa) were increased in the chronic model. Furthermore, Hsph1 combined with Vegfa was positively correlated with dilated cardiomyopathy (DCM)-induced heart failure (HF) and had high accuracy in the diagnosis of DCM-induced HF (AUC = 0.898, = 0.000).
Alas1, Atp5g1, and Ptgds were ideal biomarkers in DOX acute cardiotoxicity. However, Hsph1 and Vegfa were potential biomarkers in the myocardium in the chronic model. Our research, first, provided bioinformatics and clinical evidence for the discovery of the differences in mechanism and potential biomarkers of DOX-induced acute and chronic cardiotoxicity to find a therapeutic strategy precisely.
阿霉素（DOX）心脏毒性的机制复杂且存在争议，实验数据和临床数据之间存在各种矛盾。了解DOX诱导的急性和慢性心脏毒性分子机制的差异可能是解决这一困境的理想切入点。
分别给小鼠腹腔注射DOX（20mg/kg，一次）或（5mg/kg/周，三次）以构建急性和慢性心脏毒性模型。生存记录和超声监测心脏功能。通过RNA测序分析相应的左心室（LV）心肌组织以鉴定差异表达基因（DEG）。基因本体论（GO）、京都基因与基因组百科全书（KEGG）和基因集富集分析（GSEA）发现关键生物学过程和信号通路。将来自基因表达综合数据库（GEO）的DOX心脏毒性数据集与RNA测序相结合以鉴定共同基因。Cytoscape分析枢纽基因，并通过定量实时PCR进行验证。ImmuCo和ImmGen数据库分析枢纽基因与免疫细胞中免疫相关标志物之间的相关性。Cibersort分析免疫浸润以及枢纽基因与免疫细胞之间的相关性。逻辑回归、受试者工作特征曲线和人工神经网络分析评估枢纽基因对GEO数据集中临床数据的诊断能力。
生存曲线和超声监测表明成功构建了心脏毒性模型。在急性模型中，788个DEG富集于激活的代谢和抑制的免疫相关信号通路。三个枢纽基因（Alas1、Atp5g1和Ptgds）上调，且与免疫激活细胞集落呈负相关。然而，在慢性模型中，281个DEG表明G蛋白偶联受体（GPCR）相关信号通路是关键事件。慢性模型中有三个枢纽基因（Hsph1、Abcb1a和Vegfa）增加。此外，Hsph1与Vegfa结合与扩张型心肌病（DCM）诱导的心力衰竭（HF）呈正相关，并且在诊断DCM诱导的HF方面具有较高准确性（AUC = 0.898，P = 0.000）。
Alas1、Atp5g1和Ptgds是DOX急性心脏毒性的理想生物标志物。然而，Hsph1和Vegfa是慢性模型中心肌的潜在生物标志物。我们的研究首先为发现DOX诱导的急性和慢性心脏毒性机制差异及潜在生物标志物提供了生物信息学和临床证据，以精确找到治疗策略。
