The molecular mechanisms underlying pulmonary arterial hypertension (PAH) in congenital ventricular septal defects (VSD) are unclear. We aimed to reveal molecular pathways and potential biomarkers by multi-omics analysis in VSD-PAH. Plasma from 160 children, including 120 VSD patients with/without PAH and 40 healthy children was studied by integrated proteomics, metabolomics, and bioinformatics analyses. Proteomics (iTRAQ) identified 107 differential proteins (DPs) between patients with/without PAH including significantly increased ADIPO, DBH, ANPEP, TRF1, GP1BA and decreased GNAS in VSD-PAH. Metabolomics discovered 191 differential metabolites between patients with/without PAH including elevation of 5-HT, taurine, creatine, sarcosine, and 2-oxobutanoate and decrease of vanillylmandelic acid, 3,4-dihydroxymandelate, 15-Keto-PGF_2ᵯC;, fructose-6-phosphate , L-glutamine, dehydroascorbate, hydroxypyruvate, threonine, L-cystine, and 1-aminocyclopropane-1-carboxylate. The DPs differential proteins were validated in a new cohort of patients (N=80). Integrated analyses identified key pathways including cAMP, ECM-receptor interaction, AMPK, HIF-1, PI3K-Akt signaling pathways, and amino acid metabolisms. Increased plasma protein levels of DBH, ADIPO, and ANPEP were found to be independently associated with the occurrence of PAH with a new total risk score from these three proteins developed for clinical diagnosis. In this integrated multi-omics analysis in VSD-PAH patients, we have for the first time found that VSD-PAH patients present important differential proteins, metabolites, and key pathways. We have developed a total-risk-score (based on the plasma concentration of DBH, ANPEP and ADIPO) as predictor of development of PAH in CHD-VSD patients. Therefore, these proteins may be used as biomarkers and the new total-risk-score has significant clinical implications in the diagnosis of PAH.

中文翻译：

---

与室间隔缺损相关的肺动脉高压中的蛋白质生物标记物和风险评分：多组学和验证的整合。

先天性室间隔缺损（VSD）中的肺动脉高压（PAH）的分子机制尚不清楚。我们旨在通过VSD-PAH中的多组学分析揭示分子途径和潜在的生物标志物。通过综合蛋白质组学，代谢组学和生物信息学分析，研究了160名儿童的血浆，包括120名有/没有PAH的VSD患者和40名健康儿童。蛋白质组学（iTRAQ）在有或没有PAH的患者之间鉴定出107种差异蛋白（DP），包括ADIPO，DBH，ANPEP，TRF1，GP1BA显着增加，而VSD-PAH中的GNAS降低。代谢组学发现患有或不患有PAH的患者之间有191种差异代谢物，包括5-HT，牛磺酸，肌酸，肌氨酸和2-氧代丁酸酯的升高，香草醛酸，3,4-二羟基扁桃酸酯，15-酮基-PGF的降低_{2 ᵯC;,}果糖6-磷酸酯，L-谷氨酰胺，脱氢抗坏血酸盐，羟基丙酮酸，苏氨酸，L-胱氨酸和1-氨基环丙烷-1-羧酸酯。在新的患者队列中验证了DPs差异蛋白（N = 80）。集成分析确定了关键途径，包括cAMP，ECM-受体相互作用，AMPK，HIF-1，PI3K-Akt信号传导途径和氨基酸代谢。发现DBH，ADIPO和ANPEP的血浆蛋白水平升高与PAH的发生独立相关，这三种为临床诊断而开发的蛋白具有新的总风险评分。在VSD-PAH患者的这种综合多组学分析中，我们首次发现VSD-PAH患者具有重要的差异蛋白，代谢产物和关键途径。我们已经制定了总风险评分（根据DBH的血浆浓度，ANPEP和ADIPO）可预测CHD-VSD患者PAH的发展。因此，这些蛋白质可以用作生物标志物，新的总风险评分对PAH的诊断具有重要的临床意义。