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，10-20% 的不育男性患有无精子症，这意味着他们缺乏精子。在这两种类型中， 在许多情况下，非阻塞性​​（无阻塞）无精症是特发性的。成熟精子的产生涉及 复杂的发育过程，最终产生用于受孕的专门细胞。其中 过程包括表观遗传编程。产前精原细胞基因组规模 DNA 甲基化编程已得到充分研究，但出生后睾丸细胞的表观遗传景观尚未得到充分研究。 目前的证据表明，异常的表观遗传修饰可能是导致 特发性、非梗阻性无精症。然而，这可能与细胞组成有关，而不是与病因有关。 生精失败。为了区分这些可能性，需要进行研究以确定是否 在非梗阻性无精子症中，特定精原细胞和支持细胞的表观遗传景观发生了改变。 在目标 1 中，我们假设出生后精原细胞中异常的表观遗传景观在 无精子症是由于病因而不是整个睾丸细胞组成造成的。为了检验这个假设，我们将使用两个 无精子症遗传小鼠模型来描绘 DNA 甲基化和染色质可及性是否 妥协了。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 双突变体靶向生殖细胞区室，而第二个模型将采用 针对支持室的雄激素受体敲除 (SCARKOtm2.1)。在目标 2 中，我们假设 患有非梗阻性无精症的男性体内会存在异常的 DNA 甲基化和染色质可及性 生殖细胞或支持细胞室。为了检验这些假设，我们将产生 DNA 甲基化， 正常未分化和分化的染色质可及性和表达参考序列 精原细胞以及支持细胞，接下来我们将这些细胞的概况与来自 突变小鼠，或患有成熟停滞或仅支持细胞综合症的男性。为了这两个目的，细胞将被隔离 使用相同的富集策略以及 DNA 甲基化、染色质可及性和表达景观 对于未分化的精原细胞，将使用以下方法生成分化的精原细胞和支持细胞 NMT-seq（核小体、甲基化和转录测序），允许对 老鼠和人类。总的来说，我们将产生有关生殖细胞表观遗传景观的基础数据 和睾丸的体细胞，并确定这种景观是否在不育小鼠和男性中发生改变。