Epigenetics Landscape in the Testes of Fertile and Infertile Men

生育和不育男性睾丸的表观遗传学景观

• 批准号: 10379352
• 负责人: Mellissa Rae Wigle Mann
• 金额: $ 11.25万
• 依托单位: MAGEE-WOMEN'S RES INST AND FOUNDATION
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10379352
• 关键词: Aberrant DNA Methylation; Address; Androgen Receptor; Biopsy; Cell Compartmentation; Cells; Chromatin; Clinic; Complex; Conceptions; DNA Methylation; DNMT3B gene; DNMT3a; Data; Defect; Development; Developmental Process; Epigenetic Process; Etiology; Failure; Foundations; Genetic; Genetic Transcription; Germ Cells; Health; Hospitals; Human; Infertility; Knock-out; Knowledge; Male Infertility; Methylation; Methyltransferase; Modeling; Modification; Mus; Mutant Strains Mice; Mutation; Nucleosomes; Pilot Projects; Play; Population; Process; Production; Role; Sampling; Sertoli cell only syndrome; Somatic Cell; Spermatogenic Cell; Spermatogonia; Testing; Testis; Undifferentiated; Variant; Woman; Work; comparative; epigenetic profiling; experience; genome-wide; male; men; mouse model; mutant; postnatal; prenatal; programs; sertoli cell; sperm cell

Abstract: Pilot Project In humans, 10-20% of infertile men are azoospermic, meaning that they lack spermatozoa. Of the two types, non-obstructive (no blockage) azoospermia, in many cases, is idiopathic. Production of mature sperm involves complex developmental processes, ultimately generating specialized cells for conception. Amongst these processes includes epigenetic programming. While prenatal prospermatogonial genome-scale DNA methylation programming has been well studied, the epigenetic landscape of postnatal testicular cells has not. Current evidence suggests that aberrant epigenetic modifications may be one mechanism that leads to idiopathic, non-obstructive azoospermia. However, this may relate to cell composition rather than etiologies of spermatogenic failure. To distinguish between these possibilities, studies are required to determine whether the epigenetic landscape of specific spermatogonial and Sertoli cells is altered in non-obstructive azoospermia. In Aim 1, we hypothesize that aberrant epigenetic landscapes in postnatal spermatogonia play a role in azoospermia due to etiology rather than whole testes cell composition. To test this hypothesis, we will use two genetic mouse models with azoospermia to delineate whether DNA methylation and chromatin accessibility are compromised. The first model will eliminate all de novo methyltransferase activity using conditional Dnmt3a/Dnmt3b double mutants targeting the germ cell compartment, while the second model will employ the androgen receptor knockout (SCARKOtm2.1) targeting the Sertoli compartment. In Aim 2, we hypothesis that men with non-obstructive azoospermia will harbor aberrant DNA methylation and chromatin accessibility within either the germ cell or Sertoli compartments. To test these hypothesis, we will produce DNA methylation, chromatin accessibility and expression reference sequences from normal undifferentiated and differentiating spermatogonia as well as Sertoli cells, following which we will compare the profile of these cells to those from mutant mice, or men with maturation arrest or Sertoli cell only syndrome. For both aims, cells will be isolated using the same enrichment strategy, and DNA methylation, chromatin accessibility and expression landscapes for undifferentiated spermatogonia, differentiating spermatogonia and Sertoli cells will be generated using NMT-seq (nucleosome, methylation and transcription sequencing), allowing comparative analysis between mouse and human. Overall, we will produce foundational data about the epigenetic landscape in germ cells and somatic cells of the testes and determine whether this landscape is altered in infertile mice and men.

摘要：试点项目 在人类中，有10-20％的不育男性是Azoospermic，这意味着他们缺乏精子。在这两种类型中 在许多情况下，非目标（无障碍物）azoospermia是特发性的。成熟精子的产生涉及 复杂的发育过程，最终生成专门的细胞进行构想。其中 过程包括表观遗传编程。而产前繁荣的基因组规模DNA 已经对甲基化编程进行了充分的研究，产后睾丸细胞的表观遗传景观尚未。 当前的证据表明，异常表观遗传修饰可能是导致的一种机制 特发性，非刺激性无植物植物。但是，这可能与细胞组成有关，而不是病因 精子发生衰竭。为了区分这些可能性，需要进行研究以确定是否是否 特异性精子和塞托利细胞的表观遗传景观在非刺激性植物植物中发生了改变。 在AIM 1中，我们假设产后精子中的异常表观遗传景观在 由于病因而不是整个睾丸组成引起的植物植物。为了检验这一假设，我们将使用两个 具有Azoospermia的遗传小鼠模型描绘DNA甲基化和染色质的可及性是否为 妥协。第一个模型将使用条件消除所有从头甲基转移酶活性 DNMT3A/DNMT3B针对生殖细胞室的双突变体，而第二个模型将采用 针对Sertoli室的雄激素受体敲除（scarkotm2.1）。在AIM 2中，我们假设 具有非刺激性植物植物的男性将在内部具有异常的DNA甲基化和染色质的可及性。 生殖细胞或Sertoli室。为了检验这些假设，我们将产生DNA甲基化， 来自正常未分化和分化的染色质访问性和表达参考序列 精子性细胞和Sertoli细胞，随后我们将将这些细胞的轮廓与来自的细胞进行比较 突变小鼠，或仅成熟的男性或仅Sertoli细胞综合征。对于这两个目标，都将隔离细胞 使用相同的富集策略和DNA甲基化，染色质可及性和表达景观 对于未分化的精子，将使用分化的精子和Sertoli细胞使用 NMT-seq（核小体，甲基化和转录测序），从而可以进行比较分析 老鼠和人类。总体而言，我们将产生有关生殖细胞表观遗传景观的基础数据 睾丸的体细胞，并确定在不育小鼠和男性中是否改变了这种景观。