Congenital heart disease (CHD) with extracardiac malformations (EM) is the most common multiple malformation, resulting from the interaction between genetic abnormalities and environmental factors. Most studies have attributed the causes of CHD with EM to chromosomal abnormalities. However, multi‐system dysplasia is usually caused by both genetic mutations and epigenetic dysregulation. The epigenetic mechanisms underlying the pathogenesis of CHD with EM remain unclear. In this study, we investigated the mechanisms of imprinting alterations, including those of the Small nuclear ribonucleoprotein polypeptide N (SNRPN), PLAG1 like zinc finger 1 (ZAC1) and inositol polyphosphate‐5‐phosphatase F (INPP5F) genes, in the pathogenesis of CHD with EM. The methylation levels of SNRPN, ZAC1, and INPP5F genes were analysed by the MassARRAY platform in 24 children with CHD with EM and 20 healthy controls. The expression levels of these genes were detected by real‐time polymerase chain reaction (PCR). The correlation between methylation regulation and gene expression was confirmed using 5‐azacytidine (5‐Aza) treated cells. The methylation levels of SNRPN and ZAC1 genes were significantly increased in CHD with EM, while that of INPP5F was decreased. The methylation alterations of these genes were negatively correlated with expression. Risk analysis showed that abnormal hypermethylation of SNRPN and ZAC1 resulted in 5.545 and 7.438 times higher risks of CHD with EM, respectively, and the abnormal hypomethylation of INPP5F was 8.38 times higher than that of the control group. We concluded that abnormally high methylation levels of SNRPN and ZAC1 and decreased levels of INPP5F imply an increased risk of CHD with EM by altering their gene functions. This study provides evidence of imprinted regulation in the pathogenesis of multiple malformations.

中文翻译：

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参与先天性心脏病伴心外畸形发病机制的 SNRPN、ZAC1 和 INPP5F 基因的印记畸变。

先天性心脏病（CHD）伴心外畸形（EM）是最常见的多发性畸形，由遗传异常和环境因素相互作用引起。大多数研究将 EM 的 CHD 原因归因于染色体异常。然而，多系统发育不良通常是由基因突变和表观遗传失调引起的。具有 EM 的 CHD 发病机制的表观遗传机制仍不清楚。在这项研究中，我们调查压印改变的机制，包括那些小核核糖多肽N（的SNRPN），PLAG1像锌指1（ZAC1）和肌醇多磷酸-5-磷酸酶F（INPP5F) 基因，在 CHD 与 EM 的发病机制中。的甲基化水平SNRPN，ZAC1，并INPP5F基因通过的MassARRAY平台在24个孩子与CHD与EM和20个健康对照分析。通过实时聚合酶链反应（PCR）检测这些基因的表达水平。使用 5-氮杂胞苷 (5-Aza) 处理的细胞证实了甲基化调控与基因表达之间的相关性。的甲基化水平SNRPN和ZAC1基因显著冠心病与EM增加，而中INPP5F下降。这些基因的甲基化改变与表达呈负相关。风险分析表明异常高甲基化SNRPN和ZAC1导致具有EM，分别CHD的5.545和更高的7.438倍的风险，和异常低甲基化INPP5F比对照组的高8.38倍。我们得出的结论的那异常高甲基化水平SNRPN和ZAC1和下降的水平INPP5F意味着冠心病的风险增加EM通过改变它们的基因功能。这项研究提供了多发畸形发病机制中印迹调节的证据。