Li Zhongheng, Duolikun Maimaitiyasen, Chen Hangyu, Zhang Lei, Liu Yishuo, Li Ruining, Li Dan, Sun Lijie, Chen Long
Department of Cardiology and Institute of Vascular Medicine, Peking University Third Hospital, Beijing, China.
State Key Laboratory of Vascular Homeostasis and Remodeling, Peking University, Beijing, China.
Int J Cardiol Heart Vasc. 2025 Jul 28;60:101727. doi: 10.1016/j.ijcha.2025.101727. eCollection 2025 Oct.
While 5hmC features in cell-free DNA (cfDNA) show promise as early biomarkers for COVID-19 severity, myocardial injury, and long-term outcomes, their specific role in acute coronary syndrome (ACS) triggered shortly after COVID-19 infection remains unclear.
We generated genome-wide 5hmC profiles from plasma cfDNA using the 5hmC-Seal technique across three cohorts: ACS patients without prior COVID-19 infection (ACS, n = 16), patients experiencing ACS within 2 months post-COVID-19 (ACS2N, n = 24), and patients experiencing ACS beyond 2 months post-COVID-19 (ACS2W, n = 28). Differential 5hmC analysis, functional enrichment (GO, KEGG), immune cell deconvolution, and protein-protein interaction (PPI) network analysis were performed.
Significant differences in 5hmC profiles were identified between ACS2N and ACS patients, but not between ACS2W and ACS. Functional analysis implicated immune and inflammatory pathways. Immune infiltration analysis revealed abnormal neutrophil activation specifically in the ACS2N group. PPI network analysis pinpointed phosphodiesterase 4D (PDE4D) as a key hub gene; it was highly expressed in the ACS2N group, a finding corroborated using external datasets.
Our findings indicate that plasma cfDNA 5hmC markers can distinguish ACS occurring shortly after COVID-19 infection (ACS2N) from ACS without prior infection. We observed distinct immune dysregulation, notably abnormal neutrophil activation, in ACS2N patients. Critically, we identified PDE4D as a potential key mediator, suggesting that recent COVID-19 infection may contribute to ACS onset by abnormally upregulating PDE4D expression. This study highlights 5hmC signatures and PDE4D as potential biomarkers and therapeutic targets for post-COVID ACS.
虽然游离DNA（cfDNA）中的5-羟甲基胞嘧啶（5hmC）特征有望作为COVID-19严重程度、心肌损伤和长期预后的早期生物标志物，但其在COVID-19感染后不久引发的急性冠状动脉综合征（ACS）中的具体作用仍不清楚。
我们使用5hmC-Seal技术从血浆cfDNA中生成全基因组5hmC图谱，研究对象包括三个队列：无既往COVID-19感染的ACS患者（ACS组，n = 16）、COVID-19感染后2个月内发生ACS的患者（ACS2N组，n = 24）以及COVID-19感染后2个月以上发生ACS的患者（ACS2W组，n = 28）。进行了差异5hmC分析、功能富集（基因本体论、京都基因与基因组百科全书）、免疫细胞反卷积和蛋白质-蛋白质相互作用（PPI）网络分析。
在ACS2N组和ACS组之间发现了5hmC图谱的显著差异，但在ACS2W组和ACS组之间未发现。功能分析涉及免疫和炎症途径。免疫浸润分析显示，特别是在ACS2N组中存在异常的中性粒细胞激活。PPI网络分析确定磷酸二酯酶4D（PDE4D）为关键枢纽基因；它在ACS2N组中高表达，这一发现通过外部数据集得到了证实。
我们的研究结果表明，血浆cfDNA 5hmC标志物可以区分COVID-19感染后不久发生的ACS（ACS2N）和无既往感染的ACS。我们在ACS2N患者中观察到明显的免疫失调，尤其是异常中性粒细胞激活。至关重要的是，我们确定PDE4D为潜在的关键介质，表明近期的COVID-19感染可能通过异常上调PDE4D表达而导致ACS发病。这项研究强调了5hmC特征和PDE4D作为COVID-19后ACS的潜在生物标志物和治疗靶点。
