Single Sperm RNA signatures reveal MicroRNA biomarkers for male subfertility.

IF 4.3 3区材料科学 Q1 ENGINEERING, ELECTRICAL & ELECTRONIC ACS Applied Electronic Materials Pub Date : 2024-09-23 DOI:10.1007/s10815-024-03264-w

Masood Abu-Halima, Ulrike Fischer, Mohammad A Al Smadi, Nicole Ludwig, Anissa Acheli, Annika Engel, Hashim Abdul-Khaliq, Eckart Meese

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Abstract

Purpose: To investigate small RNA profiles in sperm, identify stable miRNA patterns unique to sperm, and assess the behavior of consistently expressed miRNAs in sperm from subfertile men compared to fertile controls.

Methods: The small RNA profiles of single sperm from four proven fertile men were analyzed using Small RNA next-generation sequencing (NGS). Subsequently, a specific set of miRNAs was validated using RT-qPCR on additional sperm samples from 65 subfertile men from an infertility clinic and 30 proven fertile men.

Results: Small RNA sequencing revealed a diverse range of sperm small RNA biotypes, including miRNAs. The mapped read percentage ranged from 22.19% for single sperm to 83.29% for enriched sperm samples used at different RNA concentrations. In single sperm, a smaller proportion of sequences were attributed to piRNAs (2.79%), miRNA (0.94%), tRNA (0.82%), and rRNA (0.47%) compared to enriched sperm samples, where piRNA (41.68%), tRNA (20.31%), miRNA (11.11%), and rRNA (6.54%) were observed. Distinct detection rates and a higher number of detected miRNAs were noted with enriched sperm samples compared to single sperm obtained using either a micromanipulator or microdissection systems. Among the identified miRNAs, 110 were consistently present in all samples. RT-qPCR revealed 15 miRNAs with increased expression and 5 miRNAs with decreased expression in sperm samples from subfertile men compared to proven fertile men. These differentially validated miRNAs were significantly correlated, either positively or negatively, with sperm count, motility, and morphology.

Conclusion: The study extensively examines small RNAs in single sperm, identifying sperm-specific miRNAs that could serve as molecular markers to distinguish between subfertile and fertile men in clinical settings.

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单精子 RNA 特征揭示了男性不育症的 MicroRNA 生物标记。

目的：研究精子中的小 RNA 图谱，确定精子特有的稳定 miRNA 模式，并评估与可育对照组相比，亚可育男性精子中持续表达的 miRNA 的行为：方法：使用小 RNA 下一代测序技术（NGS）分析了四名已证实具有生育能力男性的单个精子的小 RNA 图谱。随后，利用 RT-qPCR 对来自不孕不育诊所的 65 名亚不育男性和 30 名已证明可育男性的精子样本中的一组特定 miRNA 进行了验证：结果：小RNA测序显示了精子小RNA生物型的多样性，包括miRNA。在不同RNA浓度的富集精子样本中，映射读取率从单精子的22.19%到83.29%不等。在单个精子中，piRNA（2.79%）、miRNA（0.94%）、tRNA（0.82%）和 rRNA（0.47%）的序列比例较低，而在富集精子样本中，piRNA（41.68%）、tRNA（20.31%）、miRNA（11.11%）和 rRNA（6.54%）的序列比例较高。与使用显微机械手或显微切割系统获得的单个精子相比，富集精子样本的检测率不同，检测到的 miRNA 数量也更多。在已鉴定的 miRNA 中，有 110 个始终存在于所有样本中。RT-qPCR 发现，与已证实具有生育能力的男性相比，亚育男性精子样本中有 15 个 miRNA 表达增加，5 个 miRNA 表达减少。这些经过差异验证的 miRNA 与精子数量、活力和形态呈显著正相关或负相关：这项研究对单个精子中的小 RNA 进行了广泛研究，确定了精子特异性 miRNA，这些 miRNA 可作为分子标记，在临床上区分亚育和育龄男性。

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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