Genetics of Male Fertility Group, Unitat de Biologia Cel·lular (Facultat de Biociències), Universitat Autònoma de Barcelona, Cerdanyola del Vallès, Spain.
Sequentia Biotech SL, Barcelona, Spain.
Syst Biol Reprod Med. 2023 Jun;69(3):188-195. doi: 10.1080/19396368.2023.2183912. Epub 2023 Mar 10.
The purpose of this study is to provide novel information through Next Generation Sequencing (NGS) for the characterization of viral and bacterial RNA cargo of human sperm cells from healthy fertile donors. For this, RNA-seq raw data of poly(A) RNA from 12 sperm samples from fertile donors were aligned to microbiome databases using the GAIA software. Species of viruses and bacteria were quantified in Operational Taxonomic Units (OTU) and filtered by minimal expression level (>1% OTU in at least one sample). Mean expression values (and their standard deviation) of each species were estimated. A Hierarchical Cluster Analysis (HCA) and a Principal Component Analysis (PCA) were performed to detect common microbiome patterns among samples. Sixteen microbiome species, families, domains, and orders surpassed the established expression threshold. Of the 16 categories, nine corresponded to viruses (23.07% OTU) and seven to bacteria (2.77% OTU), among which the order and were the most abundant, respectively. HCA and PCA displayed four clusters of samples with a differentiated microbiome fingerprint. This work represents a pilot study into the viruses and bacteria that make up the human sperm microbiome. Despite the high variability observed, some patterns of similarity among individuals were identified. Further NGS studies under standardized methodological procedures are necessary to achieve a deep knowledge of the semen microbiome and its implications in male fertility.
本研究旨在通过下一代测序（NGS）为健康有生育能力的供体的人类精子细胞中的病毒和细菌 RNA 货物的特征提供新信息。为此，使用 GAIA 软件将 12 个来自有生育能力的供体的精子样本的 poly(A) RNA 的 RNA-seq 原始数据与微生物组数据库进行比对。使用操作分类单元（OTU）对病毒和细菌进行定量，并通过最小表达水平（至少在一个样本中 >1% OTU）进行过滤。估计每个物种的平均表达值（及其标准差）。进行层次聚类分析（HCA）和主成分分析（PCA）以检测样品之间的常见微生物组模式。超过既定表达阈值的 16 个微生物组物种、科、域和目。在这 16 个类别中，有 9 个对应于病毒（23.07% OTU），7 个对应于细菌（2.77% OTU），其中目和目是最丰富的。HCA 和 PCA 显示了 4 个具有不同微生物组指纹的样本聚类。这项工作代表了对构成人类精子微生物组的病毒和细菌的初步研究。尽管观察到高度的可变性，但个体之间存在一些相似模式。需要进一步进行标准化方法学程序的 NGS 研究，以深入了解精液微生物组及其对男性生育能力的影响。
