变异、缺陷的 piRNA 加工和无精子症。
Variant , Defective piRNA Processing, and Azoospermia.
机构信息
From the Division of Genetics, Oregon National Primate Research Center, Oregon Health and Science University, Beaverton (L.N., D.F.C.); the Center for Embryonic Cell and Gene Therapy, Oregon Health and Science University, Portland (D.F.C.); the Department of Growth and Reproduction (N.M., J.E.N., R.S., I.G., S.B.W., N.E.S., E.R.-D.M., N.J., K.A.) and the International Center for Research and Research Training in Endocrine Disruption of Male Reproduction and Child Health (N.M., J.E.N., R.S., I.G., S.B.W., N.E.S., E.R.-D.M., N.J., K.A.), Rigshospitalet, and the Department of Cellular and Molecular Medicine, Faculty of Health and Medical Sciences (K.A.), University of Copenhagen, Copenhagen; the Laboratory of Molecular Neurooncology, Neuroscience Institute (R.S.), and the Institute of Biology Systems and Genetic Research (I.G.), Lithuanian University of Health Sciences, Kaunas, Lithuania; the Department of Human Genetics, Donders Institute for Brain, Cognition, and Behavior (M.S.O., G.W.H.), and the Department of Obstetrics and Gynecology (G.W.H.), Radboud University Medical Center, Nijmegen, the Netherlands; Serviço de Genética, Departamento de Patologia, Faculdade de Medicina da Universidade do Porto (F.C., C.J.M.), Instituto de Investigação e Inovação em Saúde, Universidade do Porto (F.C., C.J.M., A.M.L.), and the Institute of Molecular Pathology and Immunology of the University of Porto (A.M.L.) - all in Porto, Portugal; the Andrology and In Vitro Fertilization Laboratory, Department of Surgery (Urology), University of Utah School of Medicine, Salt Lake City (K.I.A.); the Departments of Pathology and Laboratory Medicine (F.K.) and Urology (P.N.S.), Weill Cornell Medicine, New York; and the Biosciences Institute, Faculty of Medical Sciences, Newcastle University, Newcastle upon Tyne, United Kingdom (J.A.V.).
出版信息
N Engl J Med. 2021 Aug 19;385(8):707-719. doi: 10.1056/NEJMoa2028973. Epub 2021 Aug 4.
BACKGROUND
P-element-induced wimpy testis (PIWI)-interacting RNAs (piRNAs) are short (21 to 35 nucleotides in length) and noncoding and are found almost exclusively in germ cells, where they regulate aberrant expression of transposable elements and postmeiotic gene expression. Critical to the processing of piRNAs is the protein poly(A)-specific RNase-like domain containing 1 (PNLDC1), which trims their 3' ends and, when disrupted in mice, causes azoospermia and male infertility.
METHODS
We performed exome sequencing on DNA samples from 924 men who had received a diagnosis of nonobstructive azoospermia. Testicular-biopsy samples were analyzed by means of histologic and immunohistochemical tests, in situ hybridization, reverse-transcriptase-quantitative-polymerase-chain-reaction assay, and small-RNA sequencing.
RESULTS
Four unrelated men of Middle Eastern descent who had nonobstructive azoospermia were found to carry mutations in : the first patient had a biallelic stop-gain mutation, p.R452Ter (rs200629089; minor allele frequency, 0.00004); the second, a novel biallelic missense variant, p.P84S; the third, two compound heterozygous mutations consisting of p.M259T (rs141903829; minor allele frequency, 0.0007) and p.L35PfsTer3 (rs754159168; minor allele frequency, 0.00004); and the fourth, a novel biallelic canonical splice acceptor site variant, c.607-2A→T. Testicular histologic findings consistently showed error-prone meiosis and spermatogenic arrest with round spermatids of type Sa as the most advanced population of germ cells. Gene and protein expression of PNLDC1, as well as the piRNA-processing proteins PIWIL1, PIWIL4, MYBL1, and TDRKH, were greatly diminished in cells of the testes. Furthermore, the length distribution of piRNAs and the number of pachytene piRNAs was significantly altered in men carrying mutations.
CONCLUSIONS
Our results suggest a direct mechanistic effect of faulty piRNA processing on meiosis and spermatogenesis in men, ultimately leading to male infertility. (Funded by Innovation Fund Denmark and others.).
背景
P 元素诱导的软弱(testis) (PIWI)-相互作用 RNA (piRNAs) 是短的(21 到 35 个核苷酸长)和非编码的，几乎只存在于生殖细胞中，在那里它们调节转座因子的异常表达和减数分裂后基因表达。对 piRNAs 加工至关重要的是蛋白 poly(A)-特异性核糖核酸酶样域包含 1 (PNLDC1)，它修剪它们的 3'端，当在小鼠中被破坏时，导致无精子症和男性不育。
方法
我们对 924 名被诊断为非梗阻性无精子症的男性的 DNA 样本进行了外显子组测序。通过组织学和免疫组织化学测试、原位杂交、逆转录-定量-聚合酶链反应检测和小 RNA 测序分析睾丸活检样本。
结果
四名来自中东的非梗阻性无精子症的无关男性被发现携带突变：第一个患者有一个双等位基因的终止突变，p.R452Ter(rs200629089; 次要等位基因频率，0.00004)；第二个，一种新的双等位基因错义变异，p.P84S；第三个，两个复合杂合突变，包括 p.M259T(rs141903829; 次要等位基因频率，0.0007)和 p.L35PfsTer3(rs754159168; 次要等位基因频率，0.00004)；第四个，一个新的双等位基因规范剪接受体位点变异，c.607-2A→T。睾丸组织学发现一致显示易错减数分裂和精母细胞停滞，以圆精母细胞 Sa 型为最先进的生殖细胞群体。PNLDC1 的基因和蛋白表达，以及 piRNA 加工蛋白 PIWIL1、PIWIL4、MYBL1 和 TDRKH，在睾丸细胞中大大减少。此外，携带 突变的男性的 piRNAs 的长度分布和精母细胞 piRNAs 的数量显著改变。
结论
我们的结果表明，错误的 piRNA 处理对男性减数分裂和精子发生有直接的机制作用，最终导致男性不育。(由丹麦创新基金和其他机构资助)。
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