Differentially Expressed microRNAs in Peritoneal Dialysis Effluent-Derived Exosomes from the Patients with Ultrafiltration Failure.

IF 2.1 4区 生物学 Q4 GENETICS & HEREDITY
Genetics research Pub Date : 2022-08-31 eCollection Date: 2022-01-01 DOI:10.1155/2022/2276175
Weifei Wu, Xu Wu, Zhiqun Cheng, Zhenzhen Yang, Minhui Lu, Jing Cheng
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引用次数: 0

Abstract

Background: Ultrafiltration failure remains one of the most severe complications of long-term peritoneal dialysis (PD), which results in death. This study aimed to characterize the circulating exosomal microRNA (miRNA) profiles associated with ultrafiltration failure and explore its underlying mechanisms.

Methods: Exosomes were isolated from the peritoneal dialysis effluent (PDE) of patients with ultrafiltration failure or success using the ultracentrifugation method, and then transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and western blot were used for exosome characterization. After that, the isolated exosomes were sent for small RNA sequencing, and eight differentially expressed miRNAs (DE-miRNAs) were chosen for RT-qPCR validation.

Results: TEM, NTA, and western blot revealed that exosomes were successfully isolated. After sequencing, 70 DE-miRNAs involved in ultrafiltration were identified, including 41 upregulated ones and 29 downregulated ones. Functional analyses revealed that these DE-miRNAs were significantly enriched in pathways of cancer, ubiquitin-mediated proteolysis, axon orientation, and the Rap1 and Ras signaling pathways. In addition, the consistency rate of RT-qPCR and sequencing results was 75%, which indicated the relatively high reliability of the sequencing data.

Conclusions: Our findings implied that these DE-miRNAs may be potential biomarkers of ultrafiltration failure, which would help us to discover novel therapeutic targets/pathways for ultrafiltration failure in patients with end-stage renal disease.

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超滤失败患者腹膜透析流出液外泌体中差异表达的microrna
背景:超滤失败仍然是长期腹膜透析(PD)最严重的并发症之一,可导致死亡。本研究旨在描述与超滤失败相关的循环外泌体microRNA (miRNA)谱,并探讨其潜在机制。方法:采用超离心方法从超滤失败或成功患者的腹膜透析流出液(PDE)中分离外泌体,并利用透射电镜(TEM)、纳米颗粒跟踪分析(NTA)和western blot对外泌体进行表征。之后,将分离的外泌体进行小RNA测序,并选择8个差异表达的miRNAs (DE-miRNAs)进行RT-qPCR验证。结果:TEM、NTA和western blot显示成功分离外泌体。测序后,共鉴定出70个参与超滤的de - mirna,其中上调41个,下调29个。功能分析显示,这些de - mirna在癌症、泛素介导的蛋白质水解、轴突定向以及Rap1和Ras信号通路中显著富集。此外,RT-qPCR与测序结果的一致性为75%,表明测序数据具有较高的可靠性。结论:我们的研究结果表明,这些de - mirna可能是超滤衰竭的潜在生物标志物,这将有助于我们发现终末期肾病患者超滤衰竭的新治疗靶点/途径。
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来源期刊
Genetics research
Genetics research 生物-遗传学
自引率
6.70%
发文量
74
审稿时长
>12 weeks
期刊介绍: Genetics Research is a key forum for original research on all aspects of human and animal genetics, reporting key findings on genomes, genes, mutations and molecular interactions, extending out to developmental, evolutionary, and population genetics as well as ethical, legal and social aspects. Our aim is to lead to a better understanding of genetic processes in health and disease. The journal focuses on the use of new technologies, such as next generation sequencing together with bioinformatics analysis, to produce increasingly detailed views of how genes function in tissues and how these genes perform, individually or collectively, in normal development and disease aetiology. The journal publishes original work, review articles, short papers, computational studies, and novel methods and techniques in research covering humans and well-established genetic organisms. Key subject areas include medical genetics, genomics, human evolutionary and population genetics, bioinformatics, genetics of complex traits, molecular and developmental genetics, Evo-Devo, quantitative and statistical genetics, behavioural genetics and environmental genetics. The breadth and quality of research make the journal an invaluable resource for medical geneticists, molecular biologists, bioinformaticians and researchers involved in genetic basis of diseases, evolutionary and developmental studies.
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