Reference gene panel for urinary exosome-based molecular diagnostics in patients with kidney disease.

Deendayal D Mishra, Pramod K Maurya, Swasti Tiwari
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Abstract

Background: Kidney disease is a severe complication of diabetes that often leads to end-stage renal disease. Early diagnosis is crucial for prevention or delay. However, the current diagnostic methods, with their limitations in detecting the disease in its early stages, underscore the urgency and importance of finding new solutions. miRNAs encapsulated inside urinary exosomes (UEs) have potential as early biomarkers for kidney diseases. The need for reference miRNAs for accurate interpretation currently limits their translational potential.

Aim: To identify consistently expressing reference miRNAs from UEs of controls and patients with type 2 diabetesmellitus (T2DM) and biopsy-confirmed kidney diseases.

Methods: miRNA profiling was performed on UEs from 31 human urine samples using a rigorous and unbiased method. The UEs were isolated from urine samples collected from healthy individuals (n = 6), patients with T2DM (n = 13), and T2DM patients who also had kidney diseases (including diabetic nephropathy, n = 5; membranous nephropathy, n = 5; and IgA nephropathy, n = 2) through differential ultracentrifugation. After characterizing the UEs, miRNA expression profiling using microarray technology was conducted.

Results: Microarray data analysis identified 14 miRNAs that were consistently expressed in UEs from 31 human samples, representing various kidney conditions: diabetic controls, diabetic nephropathy, membrane nephropathy, IgA nephropathy, and healthy controls. Through in silico analysis, we determined that 10 of these miRNAs had significant potential to serve as reference genes in UEs.

Conclusion: We identified uniformly expressing UE miRNAs that could serve as reference genes kidney disease biomarkers.

用于肾病患者尿液外泌体分子诊断的参考基因面板。
背景:肾病是糖尿病的严重并发症,通常会导致终末期肾病。早期诊断对于预防或延缓病情至关重要。然而,目前的诊断方法在早期检测疾病方面存在局限性,这凸显了寻找新解决方案的紧迫性和重要性。封装在尿液外泌体(UEs)中的 miRNAs 有潜力成为肾脏疾病的早期生物标记物。目的:从对照组和 2 型糖尿病(T2DM)患者及活检证实的肾脏疾病患者的尿液外泌体中鉴定持续表达的参考 miRNA。方法:采用严格、无偏见的方法对 31 份人体尿液样本中的尿液外泌体进行 miRNA 分析。这些 UEs 是通过差分超速离心从健康人(6 人)、T2DM 患者(13 人)和同时患有肾脏疾病(包括糖尿病肾病,5 人;膜性肾病,5 人;IgA 肾病,2 人)的 T2DM 患者的尿液样本中分离出来的。在确定 UEs 的特征后,利用芯片技术进行了 miRNA 表达谱分析:结果:微阵列数据分析确定了 14 个 miRNA,它们在 31 个人类样本的 UEs 中持续表达,代表了不同的肾脏状况:糖尿病对照组、糖尿病肾病、膜性肾病、IgA 肾病和健康对照组。通过硅学分析,我们确定其中 10 个 miRNA 具有作为 UEs 参考基因的巨大潜力:结论:我们发现了均匀表达的 UE miRNA,它们可作为肾病生物标志物的参考基因。
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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CiteScore
3.40
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