Tang Dongdong, Li Kuokuo, Lv Mingrong, Xu Chuan, Geng Hao, Wang Chao, Cheng Huiru, He Xiaojin, Zhang Yan, Cao Yunxia
Reproductive Medicine Center, Department of Obstetrics and Gynecology, The First Affiliated Hospital of Anhui Medical University, Hefei, China.
NHC Key Laboratory of Study on Abnormal Gametes and Reproductive Tract (Anhui Medical University), Hefei, China.
Front Cell Dev Biol. 2022 May 12;10:824596. doi: 10.3389/fcell.2022.824596. eCollection 2022.
Non-obstructive azoospermia (NOA) is the most severe form of male infertility. Currently, known causative factors, including congenital and several acquired causes only account for approximately 30% of NOA cases. The causes for NOA remain unclear for most patients, which is known as idiopathic (iNOA). However, whether iNOA is due to congenital defects or acquired abnormalities is a confusing problem due to the delayed diagnosis of this frustrating condition until the childbearing age. Therefore, we collected several cases with "secondary idiopathic NOA" and detected the altered mRNAs profiles in the testicular tissues to explore the possible molecular basis. In this study, several patients with a previous history of natural pregnancy with their partners before, who were diagnosed as iNOA based on the outcomes of routine semen analysis and multiple testis biopsies now, were enrolled. Some known risk factors and genetic factors were excluded. Therefore, we defined this phenotype as "secondary idiopathic NOA." To explore the possible molecular basis of this disease, we performed mRNA expression analysis through next-generation sequencing on three cases and other three patients with obstructive azoospermia as controls. Bioinformatics analyses were conducted to assess differentially expressed genes and possible biological mechanisms involved in the disease. Quantitative real-time reverse transcription polymerase chain reaction assays were applied to confirm the results in several selected mRNAs involved in stages and metabolism of Sertoli cells. A series of mRNAs were found to be altered in testicular tissues between patients with "secondary idiopathic NOA" and controls, including 6,028 downregulated and 3,402 upregulated mRNAs. Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genome (KEGG) analyses revealed a range of GO and KEGG terms, such as cellular process involved in reproduction, protein degradation, and absorption. The present study introduces a novel classification called "secondary idiopathic NOA." We provide a global view of the altered mRNAs involved in spermatogenetic failure in these cases. Regarding the limited samples, further studies should be taken to understand this new classification.
非梗阻性无精子症（NOA）是男性不育最严重的形式。目前，已知的致病因素，包括先天性和一些后天性原因，仅占NOA病例的约30%。大多数患者的NOA病因仍不清楚，这被称为特发性（iNOA）。然而，由于这种令人沮丧的疾病直到育龄期才被诊断出来，iNOA是由于先天性缺陷还是后天异常是一个令人困惑的问题。因此，我们收集了几例“继发性特发性NOA”病例，并检测了睾丸组织中mRNA谱的变化，以探索可能的分子基础。在本研究中，纳入了几位之前曾与伴侣自然受孕、目前根据常规精液分析结果和多次睾丸活检被诊断为iNOA的患者。排除了一些已知的风险因素和遗传因素。因此，我们将这种表型定义为“继发性特发性NOA”。为了探索这种疾病可能的分子基础，我们通过下一代测序对3例患者和另外3例梗阻性无精子症患者作为对照进行了mRNA表达分析。进行了生物信息学分析，以评估差异表达基因和该疾病可能涉及的生物学机制。应用定量实时逆转录聚合酶链反应分析来确认在支持细胞阶段和代谢中涉及的几个选定mRNA的结果。发现在“继发性特发性NOA”患者和对照之间的睾丸组织中有一系列mRNA发生了改变，包括6028个下调的和3402个上调的mRNA。基因本体（GO）分析和京都基因与基因组百科全书（KEGG）分析揭示了一系列GO和KEGG术语，如参与生殖的细胞过程、蛋白质降解和吸收。本研究引入了一种名为“继发性特发性NOA”的新分类。我们提供了这些病例中参与生精失败的改变的mRNA的全局视图。鉴于样本有限，应进一步开展研究以了解这种新分类。
