Epitranscriptomic regulation of spermatogenesis and male fertility

精子发生和男性生育力的表观转录调控

基本信息

批准号：
10251021
负责人：
Wei Yan
金额：
$ 37.6万
依托单位：
LUNDQUIST INSTITUTE FOR BIOMEDICAL INNOVATION AT HARBOR-UCLA MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10251021
关键词：
3&apos Untranslated Regions Ablation Affect Alternative Splicing Animal Model Attention Biochemical Biological Assay Cell Nucleus Cells Chemicals Code Comparative Study Complex Cytoplasm Data Data Reporting Development Diagnostic Gene Expression Genes Germ Cells Health Homologous Gene Human In Vitro Initiator Codon Investigation Knockout Mice Knowledge Lead Male Contraceptive Agents Male Infertility Maps Messenger RNA Modification Molecular Mus Physiological Play Production Proteins Publishing RNA RNA Splicing Reader Regulation Reporting Reproductive Health Role Series Signal Transduction Site Somatic Cell Spermatids Spermatocytes Spermatogenesis Spermatogenic Cell Terminator Codon Testing Testis Transcript Work alpha Actin base cell type conditional knockout crosslinking and immunoprecipitation sequencing epitranscriptomics in vitro Assay in vivo in vivo Model insight interest mRNA Precursor male male fertility novel novel marker offspring protein complex sperm quality therapeutic target transcriptome transcriptome sequencing transcriptomics

项目摘要

Project Summary Reversible chemical modifications of mRNAs have recently been recognized as a major regulatory mechanism of gene expression. Among >100 various RAN chemical modifications identified so far, N6- methyladenosine (m6A) represents the most abundant one with ~3-5 m6A sites per mRNA in eukaryotic transcriptomes. Our recent work (PNAS, 2017, 115:E325) has demonstrated that ALKBH5 acts as a m6A eraser, and m6A serves as a signal for alternative splicing in the nucleus of spermatocytes and round spermatids, and for degradation in the cytoplasm of elongating and elongated spermatids. The scientific premise that proper epitranscriptomic regulation (e.g., m6A) is essential for successful spermatogenesis and male fertility prompted us to embark on investigations aiming to understand how m6A levels are controlled in developing male germ cells and what role this specific chemical modification of mRNA plays in the regulation of spermatogenesis. Based upon our published and preliminary data, we hypothesize that ALKBH4, a homolog of ALKBH5, regulates mRNA m6A levels in spermatogenic cells either as a novel eraser or a co-factor of ALKBH5. To test this hypothesis, we will first determine whether ALKBH4 functions as a novel RNA m6A demethylase or as a co-factor for ALKBH5 using biochemical analyses in vitro (Aim1). The in vitro findings will then be validated using knockout mouse models in vivo and the physiological role of ALKBH4 in the regulation of spermatogenesis will also be determined (Aim2). The proposed project will help us gain insights into the epitranscriptomic regulation of spermatogenesis and the knowledge gained would help us discover the underlying causes of poor sperm quality and male infertility.

项目概要 mRNA 的可逆化学修饰最近被认为是一种主要的调控机制。基因表达机制。迄今为止已发现的超过 100 种 RAN 化学修饰中，N6- 甲基腺苷 (m6A) 是真核生物中最丰富的一种，每个 mRNA 具有约 3-5 个 m6A 位点转录组。我们最近的工作 (PNAS, 2017, 115:E325) 证明 ALKBH5 充当 m6A 橡皮擦，m6A 作为精母细胞核中选择性剪接的信号，圆形精子细胞，以及在伸长和拉长精子细胞的细胞质中降解。这科学前提是适当的表观转录组调控（例如 m6A）对于成功至关重要精子发生和男性生育能力促使我们开始进行旨在了解的研究 m6A 水平在雄性生殖细胞发育过程中是如何控制的以及这种特定化学物质的作用 mRNA 的修饰在精子发生的调节中发挥作用。根据我们发布的和根据初步数据，我们假设 ALKBH4（ALKBH5 的同源物）调节 mRNA m6A 水平生精细胞作为新型擦除器或 ALKBH5 的辅助因子。为了检验这个假设，我们将首先确定 ALKBH4 是否作为一种新型 RNA m6A 去甲基化酶或作为使用体外生化分析的 ALKBH5 (Aim1)。然后将使用以下方法验证体外研究结果体内敲除小鼠模型以及 ALKBH4 在调节中的生理作用精子发生也将被确定（目标2）。拟议的项目将帮助我们深入了解精子发生的表观转录调控和所获得的知识将帮助我们发现精子质量差和男性不育的根本原因。