An increasing body of evidence now shows that the long-term mortality of patients with sepsis are associated with various sepsis-related immune cell defects. Alternative splicing (AS), as a sepsis-related immune cell defect, is considered as a potential immunomodulatory therapy target to improve patient outcomes. However, our understanding of the role AS plays in sepsis is currently insufficient. This study investigated possible associations between AS and the gene regulatory networks affecting immune cells. We also investigated apoptosis and AS functionality in sepsis pathophysiology. In this study, we assessed publicly available mRNA-seq data that was obtained from the NCBI GEO dataset (GSE154918), which included a healthy group (HLTY), a mild infection group (INF1), asepsis group (Seps), and a septic shock group (Shock). A total of 79 samples (excluding significant outliers) were identified by a poly-A capture method to generate RNA-seq data. The variable splicing events and highly correlated RNA binding protein (RBP) genes in each group were then systematically analyzed. For the first time, we used systematic RNA-seq analysis of sepsis-related AS and identified 1505 variable AS events that differed significantly (p <= 0.01) across the four groups. In the sepsis group, the genes related to significant AS events, such as, SHISA5 and IFI27, were mostly enriched in the cell apoptosis pathway. Furthermore, we identified differential splicing patterns within each of the four groups. Significant differences in the expression of RNA Binding Protein(RBP) genes were observed between the control group and the sepsis group. RBP gene expression was highly correlated with variant splicing events in sepsis, as determined by co-expression analysis; The expression of DDX24, CBFA2T2, NOP, ILF3, DNMT1, FTO, PPRC1, NOLC1 RBPs were significant reduced in sepsis compared to the healthy group. Finally, we constructed an RBP-AS functional network. Analysis indicated that the RBP-AS functional network serves as a critical post-transcriptional mechanism that regulates the development of sepsis. AS dysregulation is associated with alterations in the regulatory gene expression network that is involved in sepsis. Therefore, the RBP-AS expression network could be useful in refining biomarker predictions in the development of new therapeutic targets for the pathogenesis of sepsis.

中文翻译：

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败血症中失调的选择性剪接谱的全基因组鉴定和功能分析

现在越来越多的证据表明，脓毒症患者的长期死亡率与各种脓毒症相关的免疫细胞缺陷有关。选择性剪接（AS）作为一种与脓毒症相关的免疫细胞缺陷，被认为是改善患者预后的潜在免疫调节治疗靶点。然而，目前我们对 AS 在脓毒症中所起的作用的了解还不够。这项研究调查了 AS 与影响免疫细胞的基因调控网络之间可能存在的关联。我们还研究了脓毒症病理生理学中的细胞凋亡和 AS 功能。在本研究中，我们评估了从 NCBI GEO 数据集 (GSE154918) 获得的公开 mRNA-seq 数据，其中包括健康组 (HLTY)、轻度感染组 (INF1)、无菌组 (Seps) 和脓毒症组休克组（Shock）。通过 Poly-A 捕获方法鉴定了总共 79 个样本（排除显着异常值）以生成 RNA-seq 数据。然后系统地分析了每组中的可变剪接事件和高度相关的RNA结合蛋白（RBP）基因。我们首次对脓毒症相关的 AS 进行系统性 RNA 测序分析，并确定了 1505 个可变的 AS 事件，这些事件在四组中存在显着差异 (p <= 0.01)。在脓毒症组中，与重要AS事件相关的基因，如SHISA5和IFI27，大多富集在细胞凋亡途径中。此外，我们还确定了四组中每一组内的差异剪接模式。对照组和脓毒症组之间RNA结合蛋白（RBP）基因的表达存在显着差异。通过共表达分析确定，RBP 基因表达与脓毒症中的变异剪接事件高度相关；与健康组相比，脓毒症组 DDX24、CBFA2T2、NOP、ILF3、DNMT1、FTO、PPRC1、NOLC1 RBP 的表达显着降低。最后，我们构建了RBP-AS功能网络。分析表明，RBP-AS 功能网络是调节脓毒症发展的关键转录后机制。AS 失调与败血症中涉及的调节基因表达网络的改变有关。因此，RBP-AS 表达网络可用于完善生物标志物预测，从而开发脓毒症发病机制的新治疗靶点。