Impact of 3' untranslated region sequence variants in spermiogenic gene expression and infertility

3非翻译区序列变异对精子发生基因表达和不育的影响

• 批准号: 10398877
• 负责人: ANDREW W GRIMSON
• 金额: $ 54.25万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10398877
• 关键词: 3' Untranslated Regions; Address; Affect; Alleles; Bioinformatics; Biological Assay; Cell Culture Techniques; Cell Line; Cells; Chromatin; Clinical; Complex; Consequentialism; Data; Data Set; Defect; Developed Countries; Development; Epigenetic Process; Exhibits; Fertility; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genetic Transcription; Genome; Haploidy; Health; Human; In Vitro; Individual; Infertility; Length; Lesion; Male Infertility; Mastigophora; Measures; Messenger RNA; Modeling; Molecular; Morphology; Mus; Mutation; Nature; Organism; Pathogenicity; Patients; Phenotype; Phylogeny; Play; Polyadenylation; Population; Post-Transcriptional Regulation; Process; Protein Isoforms; Proteins; RNA; RNA Processing; RNA Stability; Regulation; Regulatory Element; Reporter; Reporter Genes; Ribosomes; Role; Seminal fluid; Single Nucleotide Polymorphism; Site; Source; Specific qualifier value; Sperm Count Procedure; Spermatids; Spermatocytes; Spermatogenesis; Spermiogenesis; System; Testing; Testis; Transcript; Translations; United States; Untranslated RNA; Untranslated Regions; Variant; Wild Type Mouse; asthenospermia; base; cell motility; exome; experimental study; fitness; genetic regulatory protein; mRNA Stability; mammalian genome; men; molecular phenotype; mouse model; particle; recruit; reproductive; screening; sperm cell; sperm function; trafficking; transcriptome sequencing; whole genome

Male infertility is well-recognized as a major health problem in the United States. Clinically, the major cause is poor semen quality, that is, oligozoospermia, asthenozoospermia, and/or teratozoospermia. About half of infertility cases are thought to be genetic in nature, although the molecular bases are mostly unknown. Remarkably, these sperm defects are not limited to infertile men; ~2% of all men exhibit suboptimal sperm parameters, and the sperm counts among men in many developed countries have more than halved in the past 40 years. Again, the underlying causes are unknown. Thus, it is a priority to understand the molecular basis of these disturbing increases in reproductive problems in men. With respect to genetic and epigenetic contributions to infertility, there are many stages and molecular processes that can be affected, including all stages of spermatogenesis, and roles for thousands of genes required for fertility. This project focuses on post- transcriptional gene regulation during spermiogenesis, the elaborate process by which round haploid spermatids undergo dramatic morphological development to become mature, flagellated spermatozoa with a highly compacted, inert genome. Post-transcriptional gene regulation plays a substantial role in spermiogenesis. The 3′UTR (3′ untranslated region) is central in this respect, affecting mRNA stability, translation and localization by influencing polyadenylation, access to regulatory proteins and noncoding RNAs, and loading onto ribosomes or ribonuclear protein particles (RNPs). This project seeks to understand if and how variation/mutation in the 3′UTRs of infertility patients plays a role in haploid gene expression and infertility. Aim I will determine if there are systematic or specific alterations in the post-transcriptional regulatory function of 3′UTRs in fertile versus infertile men, using 3′RNA-seq of round spermatids purified from ejaculates, and public genome sequence data. Based on these data, we will identify and prioritize variants that reside within motifs predicted bioinformatically to impact mRNA stability or other relevant attributes, focusing on genes implicated in spermiogenesis. In Aim II, candidate variants will be screened via a high-throughput in vitro system to identify those that indeed have an impact on mRNA stability and/or translation. Finally, Aim III will create mouse models of these putative pathogenic variants, and assess molecular and phenotypic effects upon reproductive parameters such as sperm number, morphology, motility, and fertility. Overall, if successful, this will be the largest effort to date to identify genetic lesions that cause infertility via disruption of post- transcriptional gene expression.

在美国，男性不育症被公认为一个主要的健康问题。临床上，主要原因是 精液质量差，即少精子症、弱精子症和/或畸精症。大约一半的人 不孕症被认为本质上是遗传的，尽管分子基础大多未知。 值得注意的是，这些精子缺陷并不局限于不育男性；大约2%的男性精子质量不佳。 参数，许多发达国家的男性精子数量在过去几年里减少了一半以上。 过去的40年里。同样，根本原因尚不清楚。因此，了解分子水平是当务之急。 这些令人不安的男性生殖问题增加的基础。关于遗传学和表观遗传学 对不孕症的贡献，有许多阶段和分子过程可以受到影响，包括所有 精子发生的阶段，以及生育所需的数千个基因的作用。本项目的重点是后 精子发生过程中转录基因的调控，即圆形单倍体的精细过程 精子细胞经历了戏剧性的形态发育，成为成熟的、有鞭毛的精子 高度紧凑、惰性的基因组。转录后基因调控在 精子发生。3‘UTR(3’非翻译区)在这方面是中心的，影响mRNA的稳定性， 通过影响多聚腺苷化、调节蛋白和非编码RNA的获得来进行翻译和定位， 并装载到核糖体或核核蛋白颗粒(RNPs)上。这个项目试图了解如果和 不孕症患者3‘端UTRs变异/突变在单倍体基因表达和不孕症中的作用 目的我将确定转录后调节功能是否有系统性或特异性的变化。 使用从精液中提纯的圆形精子细胞的3‘RNA序列，对生育男性和不育男性的3’UTRs进行比较，以及 公开基因组序列数据。基于这些数据，我们将确定驻留在 以生物信息预测的基序会影响mRNA稳定性或其他相关属性，重点放在基因上 与精子发生有关。在AIM II中，候选变异体将通过体外高通量筛选 系统识别那些确实对信使核糖核酸稳定性和/或翻译有影响的基因。最后，AIM III将 建立这些假定的致病变异的小鼠模型，并评估分子和表型效应 根据生殖参数，如精子数量、形态、活动度和生育力。总体而言，如果成功， 这将是迄今为止最大的努力，以确定通过中断后基因突变而导致不孕不育的基因损伤。 转录基因表达。