Technical considerations and review of urinary microRNAs as biomarkers for chronic kidney disease in dogs and cats.

IF 1.2 4区 农林科学 Q3 VETERINARY SCIENCES
Candice P Chu, Mary B Nabity
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引用次数: 0

Abstract

MicroRNAs (miRNAs or miRs) are small, non-coding RNAs that play a crucial role in gene regulation, making them potential biomarkers for various diseases. In the field of veterinary medicine, there is a growing interest in exploring the diagnostic and therapeutic potential of miRNAs in kidney diseases affecting dogs and cats. This review focuses on the use of urinary miRNAs as biomarkers for chronic kidney disease (CKD) in these companion animals. We introduce miRNAs, their biogenesis, and their presence in biofluids, particularly within exosomes, and discuss studies investigating miRNAs in kidney tissue and urine. We acknowledge the challenges associated with miRNA studies, including preanalytical factors such as biological variation, sample collection/processing, storage conditions, and experimental design. We highlight the importance of technical considerations, such as sample pooling, sequencing depth, multiplexing, and the various steps of the miRNA experimental workflow. Furthermore, we discuss RNA isolation methods, small RNA sequencing data analysis, and the use of quantitative reverse transcription PCR (qRT-PCR) and droplet digital PCR for verification. We emphasize the importance of internal controls, spike-ins, and normalization methods to minimize technical variation and ensure reliable results in qRT-PCR analysis. This review concludes that while urinary miRNAs hold promise as non-invasive biomarkers for CKD in dogs and cats, addressing the challenges and standardization of protocols is vital for the successful translation of this research into clinical practice.

尿微rna作为狗和猫慢性肾脏疾病生物标志物的技术考虑和综述。
MicroRNAs (miRNAs或miRs)是一种小的非编码rna,在基因调控中起着至关重要的作用,使其成为各种疾病的潜在生物标志物。在兽医学领域,探索mirna在影响狗和猫的肾脏疾病中的诊断和治疗潜力的兴趣越来越大。本文综述了在这些伴侣动物中使用尿液mirna作为慢性肾脏疾病(CKD)的生物标志物。我们介绍了mirna,它们的生物发生,以及它们在生物流体中的存在,特别是在外泌体中,并讨论了研究肾脏组织和尿液中的mirna。我们承认与miRNA研究相关的挑战,包括生物变异、样品收集/处理、储存条件和实验设计等分析前因素。我们强调了技术考虑的重要性,如样本池、测序深度、多路复用和miRNA实验工作流程的各个步骤。此外,我们讨论了RNA分离方法,小RNA测序数据分析,以及使用定量反转录PCR (qRT-PCR)和液滴数字PCR进行验证。我们强调内部控制、峰值导入和标准化方法的重要性,以最大限度地减少技术差异,并确保qRT-PCR分析结果的可靠性。这篇综述的结论是,虽然尿液mirna有望成为狗和猫CKD的非侵入性生物标志物,但解决挑战和标准化的方案对于将这项研究成功转化为临床实践至关重要。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Veterinary clinical pathology
Veterinary clinical pathology 农林科学-兽医学
CiteScore
1.70
自引率
16.70%
发文量
133
审稿时长
18-36 weeks
期刊介绍: Veterinary Clinical Pathology is the official journal of the American Society for Veterinary Clinical Pathology (ASVCP) and the European Society of Veterinary Clinical Pathology (ESVCP). The journal''s mission is to provide an international forum for communication and discussion of scientific investigations and new developments that advance the art and science of laboratory diagnosis in animals. Veterinary Clinical Pathology welcomes original experimental research and clinical contributions involving domestic, laboratory, avian, and wildlife species in the areas of hematology, hemostasis, immunopathology, clinical chemistry, cytopathology, surgical pathology, toxicology, endocrinology, laboratory and analytical techniques, instrumentation, quality assurance, and clinical pathology education.
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