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，男性不育症被广泛认为是主要的健康问题。临床上，主要原因是 精液质量差，即低脂性植物，哮喘特经疗法和/或Teratozoospermia。大约一半 尽管分子碱基大多未知，但不孕病例本质上被认为是遗传性的。 值得注意的是，这些精子缺陷不仅限于不育男人。所有男性中有2％暴露了次优的精子 参数和许多发达国家中男性的精子数量已超过一半 过去40年。同样，基本原因是未知的。那是理解分子的优先事项 这些令人不安的男性生殖问题增加的基础。关于遗传和表观遗传学 对不孕症的贡献，有许多阶段和分子过程可能会受到影响，包括所有 精子发生的阶段，以及成千上万的生育基因的作用。该项目的重点是后 精子发生过程中的转录基因调节，这是一个圆形单倍体的精细过程 精子会经历戏剧性的形态发展，成为成熟的，鞭毛的精子 高度压实，惰性基因组。转录后基因调节在 精子发生。在这方面，3'UTR（3'未翻译区域）是核心，影响mRNA稳定性， 通过影响聚腺苷酸化，调节蛋白和非编码RNA的翻译和定位， 并加载到核糖体或核糖核蛋白质颗粒（RNP）上。该项目试图了解是否和 不育患者3'UTR中的变异/突变如何在单倍体基因表达和不育中起作用。 目的，我将确定转录后调节功能是否有系统的或特定的更改 使用从射精纯净的圆形精子纯化的3'RNA-seq，肥沃的男性和不育男人的3'Utrs的 公共基因组序列数据。基于这些数据，我们将确定并确定存在于此的变体 主题在生物信息上预测会影响mRNA稳定性或其他相关属性，重点是基因 在精子发生中实施。在AIM II中，将通过高通量在体外筛选候选变体 确定确实对mRNA稳定性和/或翻译产生影响的系统。最后，AIM III将 创建这些假定的致病变异的小鼠模型，并评估分子和表型效应 在生殖参数（例如精子数，形态，运动和生育）之类的参数下。总的来说，如果成功， 迄今 转录基因表达。