Decoding the signature of sperm RNA & RNA modification of environmental stressors on the intergenerational transmission of metabolic phenotypes

解码精子 RNA 的特征

• 批准号: 10869714
• 负责人: Qi Chen
• 金额: $ 41.62万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10869714
• 关键词: Arsenic; Arsenites; Biogenesis; Bioinformatics; Biological; Cells; Code; Comparative Study; Complex; Computer software; DNA Modification Methylases; Data; Diet; Embryo; Embryonic Development; Endocrine Disruptors; Environment; Environmental Exposure; Enzymes; Exposure to; Future; Genetic Transcription; Genets; Health; Heterogeneity; High Fat Diet; Individual; Inherited; Injections; Knowledge; Lead; Ligation; Liquid Chromatography; Mammals; Masks; Maternal Exposure; Mediating; Messenger RNA; Metabolic; Metabolic Diseases; Methods; Methyltransferase; Modification; Molecular; Nature; Obesity; Oxidative Stress; Pathologic Processes; Performance; Phenotype; Physiological; Property; Protocols documentation; RNA; RNA-Directed DNA Polymerase; Ribosomes; Science; Site; Small RNA; Specificity; Testing; Therapeutic Intervention; Transfer RNA; Translating; Untranslated RNA; comparative; environmental stressor; glucose metabolism; improved; interest; intergenerational; lipid metabolism; male; metabolic phenotype; mouse model; novel; obesogenic; offspring; precision medicine; programs; ribosome profiling; sperm cell; tandem mass spectrometry; transcriptome sequencing; transcriptomics; translational applications; transmission process; tributyltin

Project Summary Emerging evidence has shown that small non-coding RNAs (sRNAs) harbor a diversity of RNA modifications. RNA modifications have the potential to store a secondary layer of labile biological information that is responsive to various environmental exposures and can modulate RNA properties such as stability and interaction potential, thus contributing to complex physiological/pathological processes. In our previous mouse model of paternal high- fat diet (HFD)-induced intergenerational inheritance, we found that tRNA-derived small RNAs (tsRNAs) and RNA methylatranserase (Dnmt2)-mediated site-specific RNA modification established a “sperm RNA code” that is required for intergenerational transmission of paternally acquired metabolic disorders (Science 2016; Nat Cell Biol 2018). These data, along with others, support an emerging concept that RNA modifications in sperm small RNAs serve as an additional layer of paternal hereditary information that can be modulated by environmental input, and is essential for regulating offspring phenotype via embryo development. These advances have set the stage to further examine whether a wider range of paternal environmental exposures, such as tributyltin (TBT) and arsenite (both are known to associate with obesity and metabolic disorders) will similarly alter sperm RNAs to confer offspring phenotype. This concerns the nature of the core sperm RNA code (i.e. a group of modified tsRNAs) shared by different exposure that is responsible for the intergenerational phenotype transmission; and also the molecular mechanism by which the modified sperm tsRNAs regulate embryo development to dictate offspring’s metabolic performance. In present project, we aim to first decipher the essential sperm tsRNAs & associated RNA modifications that responsible for programming offspring metabolic health, by comparatively studying different paternal environmental stressors (HFD, TBT & arenite exposure) with improved small RNA- seq protocol, which reduces sequencing bias by enzymatically removing RNA modifications that block reverse transcriptase and terminal adaptor ligation; we also explore the upstream regulators of the altered sperm tsRNAs, with a focus on RNA modifications enzymes (Aim 1). We will further isolate individual tsRNAs followed by RNA modification quantification using Liquid Chromatography-tandem Mass Spectrometry (LC-MS/MS), and test their function in conferring offspring phenotype by zygotic RNA injection and offspring phenotype tracking (Aim 2). Mechanistically, we will test the hypothesis whether modified sperm tsRNAs can program the metabolic state by regulating ribosome heterogeneity that control distinct translational pool of mRNAs (Aim3). In other words, we propose that environmental stressor-induced “sperm RNA code” is transformed into an “embryonic ribosome code”, which generates translational specificity to define the metabolic phenotype of offspring. Data from the proposed study may not only reveal the nature and mechanism of metabolic disorder related sperm RNA code, but also generate fundamental knowledge for future therapeutic intervention facing the obesogenic environment.

项目摘要 新的证据表明，小的非编码RNA(SRNAs)含有多种RNA修饰。 RNA修饰有可能存储第二层不稳定的生物信息 对各种环境暴露，并且可以调节诸如稳定性和相互作用势的RNA属性， 从而导致复杂的生理/病理过程。在我们之前的父性兴奋的小鼠模型中- 脂肪饮食(HFD)诱导的代际遗传，我们发现tRNA衍生的小RNA(TsRNAs)和RNA 甲基转移酶(DNMT2)介导的位点特异性RNA修饰建立了一个“精子RNA密码”，即 父系获得性代谢紊乱的代际传播所需(科学2016；NAT细胞 Biol 2018)。这些数据和其他数据一起，支持了一个新兴的概念，即精子中的RNA修饰很小 RNAs作为父系遗传信息的额外一层，可以受到环境的调节 输入，对于通过胚胎发育调节后代的表型至关重要。这些进步已经使 进一步研究更广泛的父性环境暴露，如三丁基锡(TBT) 和亚砷酸盐(已知两者都与肥胖和代谢紊乱有关)会同样改变精子的RNA 授予后代表型。这涉及到核心精子RNA编码的性质(即一组经修饰的 TsRNAs)由负责代际表型传播的不同暴露所共享；以及 此外，修饰后的精子tsRNA调节胚胎发育的分子机制也是如此。 后代的新陈代谢表现。在本项目中，我们的目标是首先破译基本精子tsRNAs& 相关的RNA修饰负责规划后代的代谢健康，通过比较 用改良的小RNA研究不同的父亲环境应激源(HFD、TBT和Arenite暴露) SEQ协议，它通过酶促移除阻碍反向的RNA修饰来减少测序偏差 转录酶和末端接头连接；我们还探索了改变的精子tsRNAs的上游调节因子， 重点是RNA修饰酶(目标1)。我们将进一步分离单个tsRNA，然后再分离RNA 用LC-MS/MS进行修饰定量，并对其进行检测 合子RNA注射和子代表型跟踪在赋予子代表型中的作用(目标2)。 从机制上讲，我们将测试这一假设，即修改后的精子tsRNAs是否可以通过以下方式来编程代谢状态 调控核糖体异质性，从而控制不同的mRNAs翻译池(Aim3)。换句话说，我们 提出环境应激源诱导的“精子RNA密码”转化为“胚胎核糖体” 密码子“，它产生翻译专一性来定义后代的代谢表型。数据来自 拟议中的研究不仅可能揭示与精子RNA密码相关的代谢障碍的性质和机制， 而且还为未来面对肥胖环境的治疗干预提供了基础知识。