Testicular piRNA profile comparison between successful and unsuccessful micro-TESE retrieval in NOA patients
Testicular piRNA profile comparison between successful and unsuccessful micro-TESE retrieval in NOA patients
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NOA 患者成功和不成功的 micro-TESE 检索的睾丸 piRNA 谱比较
DOI：
10.1007/s10815-018-1134-4
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发表时间：
2018-05-01
影响因子：
3.1
通讯作者：
Huang, Xunbin
中科院分区：
文献类型：
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作者：
Cao, Congcong;Wen, Yujiao;Huang, Xunbin
PIWI相互作用RNA（皮尔纳）是一类在哺乳动物生殖细胞中特异性表达的小分子RNA。尽管piRNA在精子发生调节中起关键作用，但在不育男性的睾丸组织中知之甚少。为探讨皮尔纳谱是否可作为临床诊断男性不育的生物标志物，本研究系统研究了显微解剖睾丸精子提取（micro-TESE）前取精成功与取精失败的非梗阻性无精子症（NOA）患者睾丸组织中piRNA的表达谱，并采用小RNA测序法（small RNA-Seq）评估piRNA的差异表达水平。对NOA患者睾丸组织进行了本体分析，以确定是否存在富集的生物过程。通过小RNA测序，从NOA患者睾丸组织中总共鉴定了18，324个智人piRNA;其中，959个piRNA在成功和不成功的精子回收组之间显著改变，其中951个睾丸piRNA显著下调，8个piRNA在精子回收不成功的NOA患者中上调。组与成功取精（SSR）组相比。出乎意料的是，发现553个睾丸piRNA在USR中完全不存在，但在SSR中显示出丰富，这表明这些piRNA可能用作micro-TESE应用的生物标志物。总共20个显著差异的piRNA（hsa-piR-20830、hsa-piR-4731、hsa-piR-6254、hsa-piR-419、hsa-piR-7152、hsa-piR-7548、hsa-piR-14195、hsa-piR-5026、hsa-piR-11482、hsa-piR-17765、hsa-piR-17102、hsa-piR-4484、hsa-piR-17260、hsa-piR-17098、hsa-piR-20511、hsa-piR-5802、hsa-piR-19121、hsa-piR-2510、hsa-piR-4745、hsa-piR-11873）以通过定量实时聚合酶链反应进一步验证RNA-Seq数据。此外，生物信息学分析显示，这些改变的piRNA参与了许多重要的生物学途径，包括细胞凋亡，细胞增殖和分化。我们的研究结果表明，NOA患者的睾丸组织中，成功和不成功的精子检索表现出不同的皮尔纳谱。这项研究为进一步阐明piRNA在精子发生中的调节作用提供了有用的资源，并为鉴定用于预测辅助生殖治疗期间NOA患者中残留生精位点的有用生物标志物提供了深刻的线索。
PIWI-interacting RNA (piRNA) is a sub-group of small RNAs about 30 nucleotides length which specifically expressed in mammalian germ cells. Although piRNAs play pivotal roles in spermatogenesis regulation, little is known in the testicular tissues of infertile men. To explore whether piRNA profile could serve as a biomarker for male infertility diagnosis in a clinic, in this study, we systematically investigated the expression profile of piRNAs in testicular tissues from the patients with non-obstructive azoospermia (NOA) between successful and unsuccessful sperm retrieval before micro-dissection testicular sperm extraction (micro-TESE).The differential expression levels of piRNAs were evaluated using small RNA-Seq method. Ontologic analyses were performed to determine the presence of enriched biological processes.A total of 18,324 Homo sapiens piRNAs were identified by small RNA-Seq from NOA patient testicular tissues; among them, 959 piRNAs were significantly altered between successful and unsuccessful sperm retrieval groups, of which 951 testicular piRNAs were significantly downregulated and 8 piRNAs were upregulated in NOA patients with unsuccessful sperm retrieval (USR) groups compared to those with successful sperm retrieval (SSR) groups, respectively. Unexpectedly, 553 testicular piRNAs were found completely absent in USR but showing abundant in SSR, which suggests that those piRNAs might serve as a biomarker for micro-TESE application. A total of 20 significantly differential piRNAs (hsa-piR-20830, hsa-piR-4731, hsa-piR-6254, hsa-piR-419, hsa-piR-7152, hsa-piR-7548, hsa-piR-14195, hsa-piR-5026, hsa-piR-11482, hsa-piR-17765, hsa-piR-17102, hsa-piR-4484, hsa-piR-17260, hsa-piR-17098, hsa-piR-20511, hsa-piR-5802, hsa-piR-19121, hsa-piR-2510, hsa-piR-4745, hsa-piR-11873) were selected to further validate the RNA-Seq data by quantitative real-time polymerase chain reaction. In addition, bioinformatic analyses revealed that those altered piRNAs were involved in many important biological pathways, including apoptosis, cell proliferation, and differentiation.Our results demonstrate that testicular tissues from NOA patients with successful and unsuccessful spermatozoa retrieval exhibit differential piRNA profiles. This study provides a useful resource to further elucidate the regulatory role of piRNAs in spermatogenesis and provides a profound clue to identify useful biomarkers for predicting residual spermatogenic loci in NOA patients during assisted reproductive treatment.