Linking Fertility-Associated Gene Polymorphisms to Aberrant Sperm Phenotypes

将生育相关基因多态性与异常精子表型联系起来

• 批准号: 9351559
• 负责人: PETER Sutovsky SUTOVSKY
• 金额: $ 43.21万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-12 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9351559
• 关键词: Affect; Agriculture; Alleles; Amino Acids; Animals; Antigens; Artificial Insemination; Biological; Biological Assay; Biological Markers; Biological Models; Breeding; Candidate Disease Gene; Cattle; Cells; Computer Simulation; Conceptions; DNA; DNA Sequence Alteration; Data; Databases; Electrons; Embryo Loss; Embryonic Development; Evaluation; Fertility; Flow Cytometry; Frequencies; Frozen Semen; Gene Targeting; Genes; Genetic; Genetic Polymorphism; Genomics; Genotype; Goals; Health; Heritability; Human; Image; In Vitro; Individual; Industry; Infertility; Knockout Mice; Life; Light; Link; Livestock; Male Infertility; Measurable; Measures; Medicine; Methods; Microscopic; Microscopy; Mutate; Mutation; Outcome; Pattern; Phenotype; Population; Pregnancy; Pregnancy loss; Proteins; Proteomics; Records; Reproductive Biology; Research; Sampling; Seminal fluid; Services; Spermatogenesis; Structure; Testing; Variant; Work; base; cellular imaging; design; embryonic protein; experimental study; falls; fertility improvement; gene product; genetic selection; genetic variant; genome sequencing; human male; improved; in vivo; innovation; insertion/deletion mutation; loss of function; loss of function mutation; male; men; microscopic imaging; novel diagnostics; phenotypic biomarker; protein function; protein structure function; reproductive; sound; sperm cell; sperm function; sperm morphology; sperm protein; sperm quality; tool; trait; whole genome

Sutovsky, Peter - PD SUMMARY The present project is directly relevant to the goals of RFA PAR-13-204, Dual Purpose with Dual Benefit: Research in Biomedicine and Agriculture Using Agriculturally Important Domestic Species. It will be the first project to systematically link the genetic traits affecting male fertility to sperm phenotypes easily measurable in a human/animal semen sample. Whole genome sequencing has identified a large number of gene polymorphisms with the potential to affect male fertility in humans and livestock. However, there is very little understanding of how these polymorphisms may affect the phenotypes of spermatozoa, their fertilizing ability and their influence on pre-embryo development and early embryo loss. We hypothesize that unique mutations/polymorphisms in genes expressed during spermatogenesis and pre- embryo development (further “fertility associated genes”) are responsible for male subfertility and for early embryo loss during pregnancy. Our goal is to link these genetic traits to sperm phenotypes measurable in a semen sample and reflected by the males’ fertility in vivo and in vitro. We chose artificial insemination (AI) bulls as model system because their genotypes are publically accessible, they have extensive fertility records from AI services, and their fertility in vivo correlates with their in vitro fertility. We have identified 3,601 candidate fertility-related genes with loss-of-function LOF) polymorphisms in the whole genome sequences of 219 bulls representing 18 breeds and performed phenotype analysis on several candidate gene products that revealed significant differences between fertile but under-performing bulls with less than satisfactory AI fertility and top bulls with highest AI fertility. Both genes also show non-synonymous polymorphisms in humans. AIM 1 of this project will employ genotyping and in silico search to identify genetic differences between fertile and subfertile bulls, including polymorphisms associated with high/low conception rates in AI, as well as those prevalent in yearling bulls that failed Breeding Soundness Evaluation (BSE). We hypothesize that fertile bulls fall on the opposite ends of fertility range due to identifiable polymorphisms in relatively few genes controlling spermatogenesis, sperm phenotype and sperm function. AIM 2 will use a combination of proteomic and cell biological approaches, including but not limited to innovative image based flow cytometry (IBFC), to link genetic fertility traits to the sperm phenotypes easily measurable in a semen sample. Based on the identification of sperm antigens expressed only or predominantly by carriers of the fertility-affecting mutations, we will develop multiplex, flow cytometry-based high throughput semen quality assays for routine use in field AI. We hypothesize that a non-synonymous change in a gene controlling spermatogenesis or sperm function will alter the sperm phenotype via change in the quantity, localization and/or function of a sperm or embryo protein encoded by this gene, thus influencing fertility of the carrier male. Altogether, we expect to identify and validate sperm phenotype biomarkers encoded by fertility associated polymorphic genes, and to improve sire management by genetic selection and objective, automated semen evaluation. This project will also yield new methods and potentially new treatments for human male and idiopathic infertility. The field of reproductive biology will be advanced through better understanding of the link between male genotype and sperm phenotype.

苏托夫斯基，彼得-PD 概括 本项目与RFA Par-13-204的目标直接相关，具有双重好处的双重目的：研究 使用农业重要的国内物种的生物医学和农业。这将是系统地的第一个项目 将影响男性生育能力的遗传特征与可在人/动物精液样本中易于测量的精子表型联系起来。 整个基因组测序已经确定了大量的基因多态性，可能会影响男性 人类和牲畜的生育能力。但是，对这些多态性如何影响 精子的表型，其施肥能力及其对胚胎前发育和早期胚胎的影响 损失。我们假设在精子发生和前表达的基因中的独特突变/多态性 胚胎发育（进一步的“生育力相关基因”）负责雄性亚生物和早期胚胎丧失 怀孕期间。我们的目标是将这些遗传特征与精液样本中可测量的精子表型联系起来 雄性在体内和体外的生育能力反映。我们选择人工授精（AI）公牛作为模型系统，因为 它们的基因型是可以公开访问的，他们拥有AI服务的广泛生育记录，并且在体内生育能力 与它们的体外生育能力相关。我们已经确定了3,601个候选生育力相关的基因 LOF）在代表18个品种并进行表型的219公牛的整个基因组序列中的多态性 对几种候选基因产品的分析，这些基因产物揭示了肥沃但表现不佳之间的显着差异 公牛少于满意的工厂AI生育能力和AI最高生育能力的顶级公牛。这两个基因也表现出非同义词 人类的多态性。 该项目的目标1将采用基因分型和计算机搜索来确定肥沃和 亚军公牛，包括与AI中高/低概念率相关的多态性，以及普遍存在的多态性 一岁的公牛，失败的繁殖健全评估（BSE）。我们假设肥沃的公牛落在相反的末端 由于可识别的多态性而导致的生育范围，在相对较少的基因控制精子发生的基因中 表型和精子功能。 AIM 2将结合蛋白质组学和细胞生物学方法的组合，包括 不限于基于创新图像的流式细胞仪（IBFC），将遗传生育特征与精子表型联系起来 在精液样品中易于测量。基于仅表达或主要通过的精子鉴定 影响生育力突变的载体，我们将发展基于流式细胞术的高吞吐量精液 在现场AI中常规使用的质量测定。我们假设基因控制的非同义变化 精子发生或精子功能将通过变化数量，定位和/或 由该基因编码的精子或胚胎蛋白的功能，因此会影响载体男性的生育能力。 总之，我们希望识别和验证由生育能力相关多态性编码的精子表型生物标志物 基因，并通过遗传选择和客观，自动精液评估来改善父亲的管理。这个项目 还将为人类和特发性不育症提供新的方法和潜在的新方法。领域 生殖生物学将通过更好地理解男性基因型与精子之间的联系来提高生殖生物学 表型。