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采用小 RNA 深度测序筛选感染 COVID-19 急性呼吸窘迫综合征患者 BALF 和血液样本中的差异表达 microRNAs。

Screening for differentially expressed microRNAs in BALF and blood samples of infected COVID-19 ARDS patients by small RNA deep sequencing.

机构信息

Genetics Research Center, The University of Social Welfare and Rehabilitation Science, Tehran, Iran.

Department of Surgery, School of Medicine, Qazvin University of Medical Sciences, Qazvin, Iran.

出版信息

J Clin Lab Anal. 2022 Nov;36(11):e24672. doi: 10.1002/jcla.24672. Epub 2022 Sep 27.

Abstract

BACKGROUND

The pandemic COVID-19 has caused a high mortality rate and poses a significant threat to the population of the entire world. Due to the novelty of this disease, the pathogenic mechanism of the disease and the host cell's response are not yet fully known, so lack of evidence prevents a definitive conclusion about treatment strategies. The current study employed a small RNA deep-sequencing approach for screening differentially expressed microRNA (miRNA) in blood and bronchoalveolar fluid (BALF) samples of acute respiratory distress syndrome (ARDS) patients.

METHODS

In this study, BALF and blood samples were taken from patients with ARDS (n = 5). Control samples were those with suspected lung cancer candidates for lung biopsy (n = 3). Illumina high-throughput (HiSeq 2000) sequencing was performed to identify known and novel miRNAs differentially expressed in the blood and BALFs of ARDS patients compared with controls.

RESULTS

Results showed 2234 and 8324 miRNAs were differentially expressed in blood and BALF samples, respectively. In BALF samples, miR-282, miR-15-5p, miR-4485-3p, miR-483-3p, miR-6891-5p, miR-200c, miR-4463, miR-483-5p, and miR-98-5p were upregulated and miR-15a-5p, miR-548c-5p, miR-548d-3p, miR-365a-3p, miR-3939, miR-514-b-5p, miR-513a-3p, miR-513a-5p, miR-664a-3p, and miR-766-3p were downregulated. On the contrary, in blood samples miR-15b-5p, miR-18a-3p, miR-486-3p, miR-486-5p, miR-146a-5p, miR-16-2-3p, miR-6501-5p, miR-365-3p, miR-618, and miR-623 were top upregulated miRNAs and miR-21-5p, miR-142a-3p, miR-181-a, miR-31-5p, miR-99-5p, miR-342-5p, miR-183-5p, miR-627-5p, and miR-144-3p were downregulated miRNAs. Network functional analysis for Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG), in ARDS patients' blood and BALF samples, showed that the target genes were more involved in activating inflammatory and apoptosis process.

CONCLUSION

Based on our results, the transcriptome profile of ARDS patients would be a valuable source for understanding molecular mechanisms of host response and developing clinical guidance on anti-inflammatory medication.

摘要

背景

新冠疫情大流行导致高死亡率,对全球人口构成重大威胁。由于这种疾病具有新颖性,其发病机制和宿主细胞的反应尚不完全清楚,因此缺乏证据无法对治疗策略做出明确结论。本研究采用小 RNA 深度测序方法筛选急性呼吸窘迫综合征(ARDS)患者血液和支气管肺泡灌洗液(BALF)样本中差异表达的 microRNA(miRNA)。

方法

本研究中,从 5 名 ARDS 患者中采集 BALF 和血液样本(n=5)。对照组为疑似肺癌患者,拟行肺活检(n=3)。采用 Illumina 高通量(HiSeq 2000)测序方法,鉴定 ARDS 患者血液和 BALF 样本中与对照组相比差异表达的已知和新型 miRNA。

结果

结果显示,血液和 BALF 样本中分别有 2234 个和 8324 个 miRNA 差异表达。在 BALF 样本中,miR-282、miR-15-5p、miR-4485-3p、miR-483-3p、miR-6891-5p、miR-200c、miR-4463、miR-483-5p 和 miR-98-5p 上调,而 miR-15a-5p、miR-548c-5p、miR-548d-3p、miR-365a-3p、miR-3939、miR-514-b-5p、miR-513a-3p、miR-513a-5p、miR-664a-3p 和 miR-766-3p 下调。相反,在血液样本中,miR-15b-5p、miR-18a-3p、miR-486-3p、miR-486-5p、miR-146a-5p、miR-16-2-3p、miR-6501-5p、miR-365-3p、miR-618 和 miR-623 是上调最明显的 miRNA,而 miR-21-5p、miR-142a-3p、miR-181-a、miR-31-5p、miR-99-5p、miR-342-5p、miR-183-5p、miR-627-5p 和 miR-144-3p 是下调最明显的 miRNA。对 ARDS 患者血液和 BALF 样本的基因本体论(GO)和京都基因与基因组百科全书(KEGG)网络功能分析表明,靶基因更多地参与激活炎症和细胞凋亡过程。

结论

基于我们的结果,ARDS 患者的转录组图谱将是理解宿主反应分子机制和开发抗炎药物临床指导的有价值的资源。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d35e/9701869/b681a1609277/JCLA-36-e24672-g008.jpg

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