Faculty of Biotechnology, Amol University of Special Modern Technologies, Amol 46156-64616, Iran.
Institute for Anatomy and Cell Biology, Medical Faculty, University of Heidelberg, Im Neuenheimer Feld 307, 69120 Heidelberg, Germany.
Int J Mol Sci. 2022 Oct 20;23(20):12570. doi: 10.3390/ijms232012570.
Non-obstructive azoospermia (NOA) is a serious cause of male infertility. The Sertoli cell responds to androgens and takes on roles supporting spermatogenesis, which may cause infertility. This work aims to enhance the genetic diagnosis of NOA via the discovery of new and hub genes implicated in human NOA and to better assess the odds of successful sperm extraction according to the individual's genotype. Whole exome sequencing (WES) was done on three NOA patients to find key genes involved in NOA. We evaluated genome-wide transcripts (about 50,000 transcripts) by microarray between the Sertoli of non-obstructive azoospermia and normal cells. The microarray analysis of three human cases with different non-obstructive azoospermia revealed that 32 genes were upregulated, and the expressions of 113 genes were downregulated versus the normal case. For this purpose, Enrich Shiny GO, STRING, and Cytoscape online evaluations were applied to predict the functional and molecular interactions of proteins and then recognize the master pathways. The functional enrichment analysis demonstrated that the biological process (BP) terms "inositol lipid-mediated signaling", "positive regulation of transcription by RNA polymerase II", and "positive regulation of DNA-templated transcription" significantly changed in upregulated differentially expressed genes (DEGs). The BP investigation of downregulated DEGs highlighted "mitotic cytokinesis", "regulation of protein-containing complex assembly", "cytoskeleton-dependent cytokinesis", and the "peptide metabolic process". Overrepresented molecular function (MF) terms in upregulated DEGs included "ubiquitin-specific protease binding", "protease binding", "phosphatidylinositol trisphosphate phosphatase activity", and "clathrin light chain binding". Interestingly, the MF analysis of the downregulated DEGs revealed overexpression in "ATPase inhibitor activity", "glutathione transferase activity", and "ATPase regulator activity". Our findings suggest that these genes and their interacting hub proteins could help determine the pathophysiologies of germ cell abnormalities and infertility.
非阻塞性无精子症（NOA）是男性不育的严重原因。支持精子发生的支持细胞对雄激素作出反应，并承担支持精子发生的作用，这可能导致不育。本工作旨在通过发现新的和与人类 NOA 相关的枢纽基因来增强 NOA 的遗传诊断，并根据个体的基因型更好地评估成功提取精子的几率。对 3 名 NOA 患者进行了全外显子组测序（WES），以找到涉及 NOA 的关键基因。我们通过微阵列评估了非阻塞性无精子症和正常细胞之间的全基因组转录物（约 50000 个转录物）。对 3 例不同非阻塞性无精子症患者的微阵列分析表明，32 个基因上调，113 个基因下调与正常病例相比。为此，应用了 Enrich Shiny GO、STRING 和 Cytoscape 在线评估来预测蛋白质的功能和分子相互作用，然后识别主要途径。功能富集分析表明，在上调的差异表达基因（DEGs）中，生物学过程（BP）术语“肌醇脂质介导的信号转导”、“RNA 聚合酶 II 转录的正调控”和“DNA 模板转录的正调控”显著改变。下调的 DEGs 的 BP 研究突出了“有丝分裂细胞分裂”、“蛋白质复合物组装的调节”、“细胞骨架依赖的细胞分裂”和“肽代谢过程”。上调的 DEGs 中过度表达的分子功能（MF）术语包括“泛素特异性蛋白酶结合”、“蛋白酶结合”、“磷酸肌醇三磷酸磷酸酶活性”和“网格蛋白轻链结合”。有趣的是，下调的 DEGs 的 MF 分析显示出“ATP 酶抑制剂活性”、“谷胱甘肽转移酶活性”和“ATP 酶调节剂活性”的过表达。我们的研究结果表明，这些基因及其相互作用的枢纽蛋白可以帮助确定生殖细胞异常和不育的病理生理学。
