Direct measurement of the male germline mutation rate using sequential sperm samples

使用连续精子样本直接测量男性种系突变率

• 批准号: 10458747
• 负责人: Gilad David Evrony
• 金额: $ 25.21万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-30 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10458747
• 关键词: Affect; Age; Aging; Back; Bar Codes; Benign; Biological Markers; Biological Specimen Banks; Blood; Characteristics; Child; Clinical; Collaborations; DNA; DNA Repair; DNA Sequence Alteration; DNA sequencing; Data; Data Analytics; Data Collection; Detection; Disease; Environmental Exposure; Environmental Risk Factor; Evolution; Experimental Designs; Family; Family Study; Fathers; Fertility; Fertilization; Foundations; Future; Genes; Genetic; Genetic Diseases; Genetic Risk; Germ-Line Mutation; Grant; Health; Human; Human Genetics; Individual; Inherited; Intellectual functioning disability; Lead; Libraries; Malignant Neoplasms; Measurement; Measures; Medical; Methodology; Methods; Molecular; Mosaicism; Mutation; Only Child; Parents; Paternal Age; Play; Population; Pregnancy; Process; Recontacts; Recording of previous events; Research; Role; Sampling; Seeds; Sperm Banks; Techniques; Technology; Time; Variant; Woman; Work; analytical method; autism spectrum disorder; base; blind; burden of illness; cohort; cost; de novo mutation; developmental disease; disorder risk; egg; genetic variant; innovation; insight; inter-individual variation; male; men; new technology; novel; novel marker; novel strategies; offspring; prospective; social; sperm cell; sperm quality; tool

PROJECT SUMMARY/ABSTRACT Most new genetic mutations in humans arise in the male germline. While most such de novo mutations are benign, collectively they drive human evolution and are responsible for an immense burden of disease— contributing to disorders such as autism, intellectual disability, and a wide range of developmental disorders. But remarkably, the rate at which mutations accumulate in the male germline, one of the most fundamental processes in human genetics, has never been directly measured. Indirect estimates of germline mutation rates have been based on family studies that identify de novo mutations in children that are not detected in their parents’ blood. These studies have shown that approximately 80% of de novo mutations arise in the male germline, and moreover, that the number of mutations transmitted by fathers via their sperm increases with paternal age. However, these ”family”-based measurements are limited by the number of children borne by any single individual, so they are only able to estimate the average mutation rate across a large number of individuals. They are also blind to mutations in sperm incapable of fertilization or viable pregnancies. Reliance on family-based measurements has therefore precluded quantification of how germline mutations accumulate with paternal age at the level of single individuals, the degree to which germline mutation rates vary across the population, and discovery of genetic and environmental factors that modify and control mutation rates. We hypothesize that directly sequencing sequential sperm samples of single individuals will define individual germline mutation rates and their inter-individual variability. In this project, we will develop a novel methodology to directly quantify, for the first time, the male germline mutation rate at the level of individuals and its variability among individuals. Aim 1 will develop a novel DNA sequencing technology (HiDEF-seq) for ultra-high fidelity detection of mosaic mutations at significantly lower cost than current methods. Aim 2 will then use HiDEF-seq to measure germline mutation rates in a novel approach whereby we will obtain sperm samples collected > 10 years apart from the same individuals in a collaboration with two of the largest sperm banks in the world. We will also develop data collection and analytic methods for use in future larger studies to define the relationships between measured mutation rates and genetic factors (e.g., variants in DNA repair/replication genes), health history, environmental exposures, fertility, and sperm quality. This work will provide a novel, scalable platform to directly measure the male germline mutation rate and, in future larger cohorts, to discover its genetic and environmental modifiers, all fundamental unknowns in human genetics. Answering these questions will provide key insights into the relationships between mutation, aging, health, and disease. This work will also provide an approach for developing germline mutation rate as a potential novel biomarker for aging, offspring disease risk, and fertility.

项目摘要/摘要 人类中的大多数新遗传突变都出现在男性种系中。虽然大多数从头突变是 良性，共同驱动人类进化，并负责巨大的疾病燃烧 - 促进自闭症，智力残疾和广泛发育障碍等疾病。 但值得注意的是，突变在男性种系中积累的速率是最多的。 人类遗传学的基本过程从未直接测量。间接估计 种系突变率是基于家庭研究，这些家庭研究鉴定了儿童的从头突变 未在父母的血液中发现。这些研究表明，大约80％的从头突变 在男性种系中出现，此外，父亲通过其精子传播的突变数量 随着父亲的年龄的增加。但是，这些基于“家庭”的测量受到的限制 任何一个人所生的孩子，因此他们只能估计一个人的平均突变率 大量个人。它们也对无法受精或可行的精子中的突变视而不见 怀孕。因此，依赖基于家庭的测量已无法定量种系 突变在单个个体水平上以父亲的年龄积聚，种系的程度 突变率在整个人群中各不相同，发现修改和改变的遗传和环境因素 控制突变率。我们假设直接测序单个个体的顺序精子样本 将定义单个种系突变率及其个体间的变异性。在这个项目中，我们将 开发一种新的方法，以首次直接量化男性种系突变率 个体的水平及其个人之间的变异性。 AIM 1将开发新的DNA测序 用于超高保真度检测镶嵌突变的技术（hidef-seq），成本明显低于 当前方法。 AIM 2然后将使用hidef-seq以一种新颖的方法来测量种系突变率 因此，我们将在合作中获得与同一个人相距十年> 10年的精子样本 拥有世界两家最大的精子库。我们还将开发数据收集和分析方法 在未来的较大研究中使用来定义测量突变率与遗传因素之间的关系 （例如，DNA修复/复制基因的变体），健康史，环境暴露，生育能力和精子 质量。这项工作将提供一个新颖，可扩展的平台，以直接测量男性种系突变率 并且，在将来的较大人群中，要发现其遗传和环境修饰符，所有的基本未知数 人类遗传学。回答这些问题将为突变之间的关系提供关键见解， 衰老，健康和疾病。这项工作还将为开发种系突变率作为一种方法 潜在的新型生物标志物，用于衰老，后代疾病风险和生育能力。