The genome integrity and epigenome of sperm from men with recurrent pregnancy loss

反复流产男性精子的基因组完整性和表观基因组

• 批准号: 10556688
• 负责人: Winifred W Mak
• 金额: $ 51.26万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10556688
• 关键词: Aberrant DNA Methylation; Address; Affect; Aging; Aniline; Animals; Area; Biological Assay; Candidate Disease Gene; Chromosomal translocation; Classification; Clinical; Couples; DNA; DNA Damage; DNA Fragmentation; DNA Methylation; Data; Data Set; Defect; Development; Diagnosis; Diagnostic; Diagnostic tests; Embryonic Development; Ensure; Epigenetic Process; Exclusion; Female; Genes; Genetic; Genome; Genomic Imprinting; Genomic Instability; Habitual Abortion; Histones; Human; Infertility; Karyotype; Knowledge; Measures; Micrococcal Nuclease; Mission; Molecular; Molecular Analysis; Mosaicism; Multiomic Data; Mutation; National Institute of Child Health and Human Development; Nested Case-Control Study; Nucleotides; Pattern; Phenotype; Play; Pregnancy; Pregnancy Complications; Pregnancy loss; Premature aging syndrome; Public Health; Recurrence; Reproductive Medicine; Research; Research Priority; Risk; Role; Seminal fluid; Spontaneous abortion; Stains; Technology; Testing; Therapeutic; United States; Work; adverse pregnancy outcome; animal data; clinical diagnostics; cohort; diagnostic strategy; early pregnancy loss; epidemiology study; epigenome; experience; experimental study; genetic analysis; genome integrity; genome-wide; genome-wide analysis; genomic locus; healthy pregnancy; imprint; improved; innovation; insight; male; men; methylome; novel; predictive modeling; sperm analysis; sperm cell; sperm quality; treatment strategy; trying to conceive; unethical

PROJECT SUMMARY/ABSTRACT: Miscarriage is the most frequent pregnancy complication, and around 750,000 to 1,000,000 miscarriages occur annually in the United States. Up to five percent of couples will experience two or more miscarriages and have recurrent pregnancy loss (RPL). RPL is a significant clinical problem as even after a diagnostic workup, 50% of couples will not find a cause for their recurrent losses and will be diagnosed with unexplained RPL (uRPL). Therefore, investigating new causes for RPL is crucial to help couples with RPL to identify a cause and, therefore, possible therapeutic options. Male factor causes of uRPL are understudied, and clinically, the only diagnostic testing performed on men with uRPL is a karyotype to exclude chromosomal translocation/inversion. Our proposal is highly significant as it addresses this major gap in our knowledge regarding paternal causes of RPL. Several lines of evidence show that abnormal sperm quality as objectively measured by high DNA fragmentation and aberrant DNA methylation in imprinted genes is associated with uRPL. This project will shift the present clinical diagnostic paradigm for RPL couples from a mainly female-centric approach to a balanced approach, including molecular analysis of sperm. Our project will involve a comprehensive analysis of sperm from a large cohort of uRPL couples to study DNA fragmentation (Aim 1), DNA methylation (Aim 2), and histone patterns (Aim 3). In Aim 1, our experiments will investigate the molecular mechanisms by which increased sperm DNA fragmentation in humans can lead to RPL by investigating the type of DNA damage occurring and the genomic loci affected. One major innovation in our approach is to uniquely study in parallel the genome of the miscarriages resulting from high DNA fragmented sperm to elucidate the underlying defect. Although sperm genomic integrity is one crucial aspect of ensuring a healthy pregnancy, another aspect is the sperm epigenome, which also plays a crucial role in pregnancy viability. Prior studies show that sperm DNA methylation at imprinted loci is perturbed in men with uRPL, and increased retention of histones in sperm from uRPL men has been observed. Building upon these observations, in aim 2, we will pursue both a candidate (imprinted genes) and an unbiased genome-wide approach to studying the sperm methylome using the latest comprehensive DNA methylation array technology. In addition, we will also leverage a prior predictive model using DNA methylation as a marker of paternal aging to assess if there is premature aging in the sperm from uRPL men. This work will identify epimutations in sperm that could play a mechanistic role in RPL. In aim 3, we will expand our epigenetic analysis to perform a genome-wide analysis of sperm histone patterns in uRPL men. Finally, we will combine our multi-omics datasets to compare and contrast the genomic loci affected. In summary, this project will contribute significantly to identifying novel male factor causes of RPL, which will ultimately lead to significant advancements in the diagnosis and treatment of couples with RPL.

项目摘要/摘要：流产是最常见的妊娠并发症，大约 美国每年发生 750,000 至 1,000,000 起流产。高达百分之五的夫妇会 经历两次或两次以上流产并有复发性流产 (RPL)。 RPL是一个重要的临床 问题在于，即使在进行诊断检查后，50% 的夫妇也无法找到其经常性损失的原因，并且 将被诊断为不明原因的 RPL (uRPL)。因此，调查 RPL 的新原因对于帮助患者至关重要 与 RPL 结合以确定病因，从而确定可能的治疗方案。男性因素导致uRPL 尚未得到充分研究，并且在临床上，对患有 uRPL 的男性进行的唯一诊断测试是核型 排除染色体易位/倒位。我们的提案非常重要，因为它解决了这一重大差距 据我们所知，RPL 的父亲原因。多项证据表明，异常精子 通过印记基因中的高 DNA 碎片和异常 DNA 甲基化客观衡量质量 与 uRPL 相关。该项目将改变目前 RPL 夫妇的临床诊断模式 主要以女性为中心的方法采取平衡的方法，包括精子的分子分析。我们的项目将 涉及对一大群 uRPL 夫妇的精子进行全面分析，以研究 DNA 碎片 （目标 1）、DNA 甲基化（目标 2）和组蛋白模式（目标 3）。在目标 1 中，我们的实验将研究 人类精子 DNA 碎片增加导致 RPL 的分子机制 研究发生的 DNA 损伤类型和受影响的基因组位点。我们的一项重大创新 方法是并行研究由高 DNA 片段化导致的流产的基因组 精子来阐明潜在的缺陷。尽管精子基因组完整性是确保精子基因组完整性的一个重要方面 健康怀孕，另一个方面是精子表观基因组，它对怀孕也起着至关重要的作用 生存能力。先前的研究表明，在患有 uRPL 的男性中，印记位点的精子 DNA 甲基化受到干扰，并且 已观察到 uRPL 男性精子中组蛋白的保留增加。在此基础上 根据观察结果，在目标 2 中，我们将追求候选基因（印记基因）和无偏见的全基因组 使用最新的综合 DNA 甲基化阵列技术研究精子甲基化组的方法。 此外，我们还将利用先前的预测模型，使用 DNA 甲基化作为父亲衰老的标志 评估 uRPL 男性的精子是否存在过早衰老。这项工作将鉴定表观突变 精子可能在 RPL 中发挥机械作用。在目标 3 中，我们将扩展我们的表观遗传分析以执行 uRPL 男性精子组蛋白模式的全基因组分析。最后，我们将结合我们的多组学 数据集来比较和对比受影响的基因组位点。总而言之，该项目将贡献 显着有助于识别 RPL 的新男性因素，这最终将导致显着的 RPL 夫妇诊断和治疗方面的进展。