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是一个主要的健康问题，其主要原因是。 精液质量差，即少精症、弱精症和/或畸形精子症。 尽管分子基础大多未知，但不孕症病例被认为本质上是遗传性的。 值得注意的是，这些精子缺陷不仅限于不育男性；约 2% 的男性表现出不理想的精子。 参数，许多发达国家男性的精子数量减少了一半以上 过去 40 年来，根本原因尚不清楚，因此，首先要了解分子。 这些令人不安的男性生殖问题增加的基础是遗传和表观遗传。 对不孕不育的贡献，有许多阶段和分子过程可能受到影响，包括所有 精子发生的各个阶段以及生育所需的数千个基因的作用该项目重点关注精子发生后的过程。 精子发生过程中的转录基因调控，这是单倍体形成的复杂过程 精子细胞经历戏剧性的形态发育，成为成熟的、有鞭毛的精子， 高度紧凑的惰性基因组在转录后基因调控中发挥着重要作用。 3'UTR（3'非翻译区）在这方面至关重要，影响 mRNA 的稳定性， 通过影响多聚腺苷酸化、调节蛋白和非编码 RNA 的获取来进行翻译和定位， 该项目旨在了解是否和加载到核糖体或核糖核蛋白颗粒（RNP）上。 不孕症患者 3'UTR 的变异/突变如何在单倍体基因表达和不孕症中发挥作用。 目标 我将确定转录后调节功能是否存在系统或特定的改变 使用从精液中纯化的圆形精子细胞的 3'RNA-seq 分析生育男性与不育男性的 3'UTR，以及 根据这些数据，我们将识别并优先考虑其中存在的变异。 以生物信息学方式预测影响 mRNA 稳定性或其他相关属性的基序，重点关注基因 在 Aim II 中，将通过高通量体外筛选候选变体。 系统来识别那些确实对 mRNA 稳定性和/或翻译有影响的因素。最后，Aim III 将会。 创建这些假定致病变异的小鼠模型，并评估分子和表型效应 根据精子数量、形态、活力和生育力等生殖参数，如果成功的话， 这将是迄今为止最大的努力，旨在识别通过破坏后遗症而导致不孕的基因损伤。 转录基因表达。