Transcriptional consequences of impaired immune cell responses induced by cystic fibrosis plasma characterized via dual RNA sequencing
Transcriptional consequences of impaired immune cell responses induced by cystic fibrosis plasma characterized via dual RNA sequencing
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DOI：
10.1186/s12920-019-0529-0
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发表时间：
2019-05-22
影响因子：
2.7
通讯作者：
Levy, Hara
中科院分区：
文献类型：
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作者：
Ideozu, Justin E.;Rangaraj, Vittobai;Levy, Hara
背景在囊性纤维化（CF）中，由功能失调的CF跨膜电导调节（CFTR）基因驱动的免疫细胞反应受损可能决定疾病的严重程度，但临床异质性仍然是主要的治疗挑战。 CF 免疫反应受损的分子机制特征可能会揭示具有治疗潜力的新靶点。因此，我们利用同步 RNA 测序来识别差异表达的基因、转录本和 miRNA，这些基因、转录本和 miRNA 表征了 CF 及其表型引发的免疫反应受损的特征。方法用 CF 患者 (n = 9) 和健康对照 (n = 3) 的血浆刺激从健康供体中提取的外周血单核细胞 (PBMC)。将 PBMC 在 37 摄氏度、5% CO2 下培养（1x10(5) 细胞/孔）9 小时。培养后，从每个样本中提取总RNA，并用于同步总RNA和miRNA测序。结果分析CF患者和健康对照血浆诱导的外周血单核细胞的表达特征，发现CF中与HC相比差异表达的151个基因、154个个体转录物和41个miRNA，而285个基因、241个个体转录物和7个miRNA的表达特征因CF表型而不同。受 CF 影响的主要免疫途径包括粒细胞粘附、血细胞渗出信号传导和 IL17 信号传导，而受 CF 表型影响的免疫途径包括自然杀伤细胞信号传导和 B 淋巴细胞中的 PI3K 信号传导。上游调节因子分析表明，CF 导致 CCL5、NF-B 和 IL1A 失调，而 TREM1 和 TP53 调节因子失调与 CF 表型相关。 5 个 miRNA 显示出与 CF 相关受损免疫通路相关的 3 个靶基因的反向表达模式，而 2 个 miRNA 显示出与与 CF 表型相关的失调免疫通路相关的两个靶基因的反向表达模式。结论我们的结果表明 miRNA 和单个转录物变体是导致 CF 中免疫细胞反应受损的相关分子靶标。
BackgroundIn cystic fibrosis (CF), impaired immune cell responses, driven by the dysfunctional CF transmembrane conductance regulator (CFTR) gene, may determine the disease severity but clinical heterogeneity remains a major therapeutic challenge. The characterization of molecular mechanisms underlying impaired immune responses in CF may reveal novel targets with therapeutic potential.Therefore, we utilizedsimultaneous RNA sequencing targeted at identifying differentially expressed genes, transcripts, and miRNAs that characterize impaired immune responses triggered by CF and its phenotypes.MethodsPeripheral blood mononuclear cells (PBMCs) extracted from a healthy donor were stimulated with plasma from CF patients (n=9) and healthy controls (n=3). The PBMCs were cultured (1x10(5) cells/well) for 9h at 37 degrees C in 5% CO2. After culture, total RNA was extracted from each sample and used for simultaneous totalRNAand miRNA sequencing.ResultsAnalysis of expression signatures from peripheral blood mononuclear cells induced by plasma of CF patients and healthy controls identified 151 genes, 154 individual transcripts, and 41 miRNAs differentially expressed in CF compared to HC while the expression signatures of 285 genes, 241 individual transcripts, and seven miRNAs differed due to CF phenotypes. Top immune pathways influenced by CF included agranulocyte adhesion, diapedesis signaling, and IL17 signaling, while those influenced by CF phenotypes included natural killer cell signaling and PI3K signaling in B lymphocytes. Upstream regulator analysis indicated dysregulation of CCL5, NF-B and IL1A due to CF while dysregulation of TREM1 and TP53 regulators were associated with CF phenotype. Five miRNAs showed inverse expression patterns with three target genes relevant in CF-associated impaired immune pathways while two miRNAs showed inverse expression patterns with two target genes relevant to a dysregulated immune pathway associated with CF phenotypes.ConclusionsOur results indicate that miRNAs and individual transcript variants are relevant molecular targets contributing to impaired immune cell responses in CF.