Department of Pulmonary and Critical Care Medicine, Beijing Anzhen Hospital, Capital Medical University, Beijing, China.
National Center for Respiratory Medicine; State Key Laboratory of Respiratory Health and Multimorbidity; National Clinical Research Center for Respiratory Diseases; Institute of Respiratory Medicine, Chinese Academy of Medical Sciences; Department of Pulmonary and Critical Care Medicine, Center of Respiratory Medicine, China-Japan Friendship Hospital, Beijing, China.
Respir Res. 2024 Nov 28;25(1):418. doi: 10.1186/s12931-024-03036-1.
Identification and validation of potential biomarkers could facilitate the identification of pulmonary arterial hypertension (PAH) and thus aid to study their roles in the disease process.
We used the isobaric tag for relative and absolute quantitation approaches to compare the protein profiles between the serum of PAH patients and the controls. Bioinformatics analyses and enzyme-linked immunosorbent assay (ELISA) identification of PAH patients and the controls were performed to identify the potential biomarkers. The receiver operating characteristic curve (ROC) analysis was used to evaluate the diagnostic performance of these potential biomarkers. Mendelian randomization (MR) analysis further clarified the relationship between the potential biomarkers and PAH. Additionally, the expression levels of the potential biomarkers were further validated in two PAH animal models (monocrotaline-PH and Sugen5416 plus hypoxia-PH) using ELISA and reverse transcription-quantitative PCR (RT-qPCR).
We identified significant changes in three proteins including heparanase (HPSE), gelsolin (GSN), and hepatocyte growth factor activator (HGFA) in PAH patients. The ROC analysis showed that the areas under the curve of HPSE, GSN, and HGFA in differentiating PAH patients from controls were 0.769, 0.777, and 0.964, respectively. HGFA was correlated with multiple parameters of right ventricular functions in PAH patients. Besides proteomic analysis, we also used MR method to demonstrate the causal link between genetically reduced HGFA levels and an increased risk of PAH. In subsequent validation study in PAH animal models, the mRNA expression levels of HGFA in the lung tissues were significantly lower in PAH rat models than in controls. In the rat models, serum levels of HGFA were lower compared to the control group and showed a negative correlation with right ventricular systolic pressure.
The study demonstrated that HGFA might be a promising biomarker for noninvasive detection of PAH.
识别和验证潜在的生物标志物可以促进肺动脉高压（PAH）的识别，从而有助于研究它们在疾病过程中的作用。
我们使用相对和绝对定量的同位标签技术比较了 PAH 患者和对照组血清中的蛋白质谱。通过生物信息学分析和酶联免疫吸附试验（ELISA）对 PAH 患者和对照组进行鉴定，以确定潜在的生物标志物。利用受试者工作特征曲线（ROC）分析评估这些潜在生物标志物的诊断性能。孟德尔随机化（MR）分析进一步阐明了潜在生物标志物与 PAH 之间的关系。此外，还使用 ELISA 和逆转录定量 PCR（RT-qPCR）在两种 PAH 动物模型（野百合碱-PH 和 Sugen5416 加低氧-PH）中进一步验证了潜在生物标志物的表达水平。
我们发现 PAH 患者中有三种蛋白质（包括肝素酶[HPSE]、凝溶胶蛋白[GSN]和肝细胞生长因子激活剂[HGFA]）发生了显著变化。ROC 分析显示，HPSE、GSN 和 HGFA 区分 PAH 患者和对照组的曲线下面积分别为 0.769、0.777 和 0.964。HGFA 与 PAH 患者右心功能的多个参数相关。除了蛋白质组学分析，我们还使用 MR 方法证明了 HGFA 水平降低与 PAH 风险增加之间的因果关系。在随后的 PAH 动物模型验证研究中，HGFA 在肺组织中的 mRNA 表达水平在 PAH 大鼠模型中明显低于对照组。在大鼠模型中，血清 HGFA 水平低于对照组，与右心室收缩压呈负相关。
该研究表明，HGFA 可能是一种有前途的非侵入性检测 PAH 的生物标志物。
