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

基本信息

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
项目类别：
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

项目摘要

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.

彼得·苏托夫斯基 - P.D. 概括目前的项目与 RFA PAR-13-204 的目标直接相关，双重目的和双重利益：研究利用农业重要国内物种的生物医学和农业这将是第一个系统地进行的项目。将影响男性生育能力的遗传特征与可在人类/动物精液样本中轻松测量的精子表型联系起来。全基因组测序已鉴定出大量可能影响男性的基因多态性然而，人们对这些多态性如何影响人类和牲畜的生育能力知之甚少。精子的表型、受精能力及其对胚胎前发育和早期胚胎的影响我们追求精子发生和预表达过程中基因的独特突变/多态性。胚胎发育（进一步的“生育相关基因”）是导致男性生育力低下和早期胚胎丢失的原因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。通过雄性体内和体外的生育能力来反映我们选择人工授精（AI）公牛作为模型系统是因为。他们的基因型是公开的，他们有来自人工智能服务的大量生育记录，以及他们的体内生育能力我们已经鉴定出 3,601 个功能丧失的候选生育相关基因。 LOF) 代表 18 个品种和表现型的 219 头公牛的全基因组序列的多态性对几个候选基因产物的分析揭示了可育但表现不佳之间的显着差异 AI 生育力较差的公牛和 AI 生育力最高的公牛这两个基因也表现出非同义。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识别可育和可育之间的遗传差异生育力低下的公牛，包括与人工智能中高/低受孕率相关的多态性，以及在未通过育种健全性评估 (BSE) 的一岁公牛我们发现，可育公牛的情况正好相反。由于控制精子发生、精子的相对较少的基因中可识别的多态性，生育范围的范围 AIM 2 将结合蛋白质组学和细胞生物学方法，包括但不限于表型和精子功能。不限于创新的基于图像的流式细胞术 (IBFC)，将遗传生育性状与精子表型联系起来基于仅或主要表达的精子抗原的鉴定，可以在精液样本中轻松测量。作为影响生育能力的突变的携带者，我们将开发基于流式细胞术的多重高通量精液现场人工智能常规使用的质量分析我们认为基因控制中的非同义变化。精子发生或精子功能将通过数量、定位和/或的变化来改变精子表型该基因编码的精子或胚胎蛋白的功能，从而影响携带者男性的生育能力。总而言之，我们期望识别和验证由生育相关多态性编码的精子表型生物标志物该项目通过遗传选择和客观、自动化的精液评估来改善父系管理。还将产生针对人类男性和特发性不育症领域的新方法和潜在新疗法。通过更好地了解男性基因型和精子之间的联系，生殖生物学将得到进步表型。