Changes in specific circulating RNA (circRNA) expressions can serve as diagnostic noninvasive biomarkers for prostate cancer (PCa). However, there are still unmet needs, such as unclear types and roles of circRNAs, PCa detection in benign prostatic hyperplasia (BPH) by unstandardized methods, and limitations of sample volume capacity and low circRNA concentrations. This study reports a simple and rapid circRNA enrichment and isolation technique named “HAZIS-CirR” for the analysis of urinary circRNAs. The method utilizes homobifunctional hydrazides with amine-modified zeolite and polyvinylidene fluoride (PVDF) syringe filtration for combining electrostatic and covalent coupling and size-based filtration, and it offers instrument-free isolation of circRNAs in 20 min without volume limitation, thermoregulation, and lysis. HAZIS-CirR has high capture efficiency (82.03%–92.38%) and a 10-fold more sensitive detection limit (20 fM) than before enrichment (200 fM). The clinical utility of HAZIS-CirR is confirmed by analyzing circulating mRNAs and circulating miRNAs in 89 urine samples. Furthermore, three miRNA panels that differentiate PCa from BPH and control, PCa from control, and BPH from control, respectively, are established by comparing miRNA levels. HAZIS-CirR will be used as an optimal and established method for the enrichment and isolation of circRNAs as diagnostic, prognostic, and predictive biomarkers in human cancers.

中文翻译：

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使用同双功能酰肼富集和分离尿液循环 RNA 的新型平台


特定循环 RNA (circRNA) 表达的变化可以作为前列腺癌 (PCa) 的诊断性非侵入性生物标志物。然而，仍然存在未满足的需求，例如circRNA的类型和作用不清楚、良性前列腺增生（BPH）中PCa的检测方法不标准化、样本容量和低circRNA浓度的限制等。本研究报告了一种简单快速的 circRNA 富集和分离技术，名为“HAZIS-CirR”，用于分析尿液 circRNA。该方法利用同双功能酰肼、胺改性沸石和聚偏二氟乙烯 (PVDF) 注射器过滤，将静电和共价偶联以及基于尺寸的过滤相结合，无需仪器即可在 20 分钟内分离 circRNA，无需体积限制、温度调节和裂解。 。 HAZIS-CirR 具有较高的捕获效率 (82.03%–92.38%)，检测限 (20 fM) 比富集前 (200 fM) 高 10 倍。通过分析 89 个尿液样本中的循环 mRNA 和循环 miRNA，证实了 HAZIS-CirR 的临床实用性。此外，通过比较 miRNA 水平，建立了三个 miRNA 组，分别区分 PCa 与 BPH 和对照、PCa 与对照、以及 BPH 与对照。 HAZIS-CirR 将用作富集和分离 circRNA 作为人类癌症诊断、预后和预测生物标志物的最佳且成熟的方法。