Department of Pediatrics (Cardiology) University of Washington Seattle WA.
Center for Developmental Biology and Regenerative Medicine Seattle Children's Research Institute Seattle WA.
J Am Heart Assoc. 2022 Sep 6;11(17):e025864. doi: 10.1161/JAHA.122.025864. Epub 2022 Aug 24.
Background The systemic inflammation that occurs after exposure to cardiopulmonary bypass (CPB), which is especially severe in neonatal patients, is associated with poorer outcomes and is not well understood. In order to gain deeper insight into how exposure to bypass activates inflammatory responses in circulating leukocytes, we studied changes in microRNA (miRNA) expression during and after exposure to bypass. miRNAs are small noncoding RNAs that have important roles in modulating protein levels and function of cells. Methods and Results We performed miRNA-sequencing on leukocytes isolated from neonatal patients with CPB (n=5) at 7 time points during the process of CPB, including before the initiation of bypass, during bypass, and at 3 time points during the first 24 hours after weaning from bypass. We identified significant differentially expressed miRNAs using generalized linear regression models, and miRNAs were defined as statistically significant using a false discovery rate-adjusted <0.05. We identified gene targets of these miRNAs using the TargetScan database and identified significantly enriched biological pathways for these gene targets. We identified 54 miRNAs with differential expression during and after CPB. These miRNAs clustered into 3 groups, including miRNAs that were increased during and after CPB (3 miRNAs), miRNAs that decreased during and after CPB (10 miRNAs), and miRNAs that decreased during CPB but then increased 8 to 24 hours after CPB. A total of 38.9% of the target genes of these miRNAs were significantly differentially expressed in our previous study. miRNAs with altered expression levels are predicted to significantly modulate pathways related to inflammation and signal transduction. Conclusions The unbiased profiling of the miRNA changes that occur in the circulating leukocytes of patients with bypass provides deeper insight into the mechanisms that underpin the systemic inflammatory response that occurs in patients after exposure to CPB. These data will help the development of novel treatments and biomarkers for bypass-associated inflammation.
体外循环（CPB）后发生的全身炎症反应在新生儿患者中尤为严重，与较差的预后相关，但目前尚不清楚其发生机制。为了更深入地了解体外循环暴露如何激活循环白细胞中的炎症反应，我们研究了体外循环期间和之后 miRNA（miRNA）表达的变化。miRNA 是一种小的非编码 RNA，在调节细胞蛋白质水平和功能方面发挥着重要作用。
我们对接受 CPB 的新生儿患者（n=5）的白细胞进行了 miRNA 测序，在 CPB 过程中，包括在开始体外循环之前、体外循环期间以及从体外循环脱机后的前 24 小时内的 7 个时间点进行了 miRNA 测序。我们使用广义线性回归模型确定了显著差异表达的 miRNA，使用错误发现率校正<0.05 来定义 miRNA 具有统计学意义。我们使用 TargetScan 数据库确定了这些 miRNA 的基因靶标，并确定了这些基因靶标显著富集的生物学途径。我们确定了 54 个在 CPB 期间和之后差异表达的 miRNA。这些 miRNA 聚类为 3 组，包括在 CPB 期间和之后增加的 miRNA（3 个）、在 CPB 期间和之后减少的 miRNA（10 个）以及在 CPB 期间减少但在 CPB 后 8 至 24 小时增加的 miRNA。我们之前的研究中，这些 miRNA 的靶基因有 38.9%表达显著差异。表达水平改变的 miRNA 被预测会显著调节与炎症和信号转导相关的途径。
对接受体外循环的患者循环白细胞中发生的 miRNA 变化进行的无偏分析，提供了对体外循环暴露后患者发生全身炎症反应的机制的更深入了解。这些数据将有助于开发与体外循环相关炎症相关的新治疗方法和生物标志物。
