ERK-mediated regulation of non-coding RNAs during development and disease

发育和疾病过程中 ERK 介导的非编码 RNA 调节

• 批准号: 10487399
• 负责人: Swathi Arur
• 金额: $ 40.5万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-10 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10487399
• 关键词: Area; Arginine; Biogenesis; Biological Assay; Biology; Birth; CRISPR/Cas technology; Caenorhabditis elegans; Cell Culture System; Cells; Congenital Abnormality; Development; Disease; Embryo; Embryonic Development; Endometrial Carcinoma; Event; Generations; Growth Factor; Human; Human Biology; Image; Infertility; Investigation; Knowledge; Light; Link; Malignant Neoplasms; Mammalian Cell; Mammals; Mediating; Meiosis; Methods; Methylation; Modeling; Morphogenesis; Oocytes; Phosphorylation; Population; Process; Proteome; Proteomics; Puberty; RNA; RNA Degradation; Regulation; Reproduction; Research; Role; Signal Transduction; Small RNA; Sterility; Untranslated RNA; Work; genome editing; human female; in vivo Model; mouse model; next generation sequencing; oocyte quality; sperm cell; transcriptome

PROJECT SUMMARY Successful reproduction through the fusion of the sperm and oocyte is essential for the perpetuation of species. In human females, oocytes complete meiosis I at birth, and enter a long period of meiotic II arrest until onset of meiotic maturation at puberty. Because the oocytes are quiescent and arrested during this period, RNAs are loaded into the developing oocytes prior to the arrest and these RNAs are critical for early embryonic development. Mechanisms that regulate generation and protection (from degradation) of maternal RNAs during the long meiotic arrest as well as mechanisms that regulate the degradation of these RNAs in the embryo remain an active area of investigation. Our work in C. elegans and work from mammalian models in the past few years turned the light on regulation of the maternal transcriptome which dictates oocyte quality and impacts progeny development. Specifically, we uncovered a direct link between RAS/ERK growth factor signaling and the small RNA biogenesis factors Dicer1, Drosha and DIS3 (an RNA exosomal component) which regulates distinct populations of small non-coding RNAs and thus the maternal transcriptome and proteome. We propose a model wherein ERK-mediated phosphorylation of Dicer1 (and a subsequent arginine methylation of Dicer1), phosphorylation of Drosha and DIS3 results in a regulatory circuit that fine tunes the generation of small non-coding RNAs in specific subsets and regulates the maternal and zygotic transcriptome and proteome. We investigate this model in vivo during oocyte development and oocyte-to-embryo transition using a combination of live imaging, next generation sequencing, single oocyte sequencing, mass spectrometric and proteomic methods, CRISPR Cas9 genome editing and cell biological assays. We find that Dicer1, Drosha and Dis3 are phosphorylated in mammals as well. Additionally, we identified arginine methylation of Dicer1 adjacent to the phosphorylation event in mammalian cell culture system. Given their conserved role in RNA biology, reproduction and their aberrations associated with cancer onset and progression, we expect this work to have direct relevance to human biology.

项目概要 通过精子和卵母细胞融合的成功繁殖对于物种的延续至关重要。 在人类女性中，卵母细胞在出生时完成减数分裂 I，并进入长期的减数分裂 II 停滞期，直到 减数分裂在青春期开始成熟。因为卵母细胞在此期间处于静止和停滞状态， RNA 在逮捕前被加载到发育中的卵母细胞中，这些 RNA 对于早期发育至关重要。 胚胎发育。调节生成和保护（防止退化）的机制 长期减数分裂停滞期间的母体 RNA 以及调节这些降解的机制 胚胎中的 RNA 仍然是一个活跃的研究领域。我们在秀丽隐杆线虫和哺乳动物中的工作 过去几年的模型揭示了决定卵母细胞的母体转录组的调节 质量并影响后代发育。具体来说，我们发现了 RAS/ERK 生长之间的直接联系 因子信号转导和小 RNA 生物发生因子 Dicer1、Drosha 和 DIS3（RNA 外泌体成分） 它调节不同的小非编码 RNA 群体，从而调节母体转录组和 蛋白质组。我们提出了一个模型，其中 ERK 介导的 Dicer1 磷酸化（以及随后的精氨酸 Dicer1 的甲基化）、Drosha 和 DIS3 的磷酸化会形成一个调节电路，可微调 在特定亚群中生成小非编码 RNA 并调节母体和合子转录组 和蛋白质组。我们在卵母细胞发育和卵母细胞向胚胎转变过程中体内研究了该模型 结合使用实时成像、下一代测序、单卵母细胞测序、质谱 和蛋白质组学方法、CRISPR Cas9 基因组编辑和细胞生物学测定。我们发现 Dicer1、Drosha 和 Dis3 在哺乳动物中也被磷酸化。此外，我们还发现了 Dicer1 的精氨酸甲基化 与哺乳动物细胞培养系统中的磷酸化事件相邻。鉴于它们在 RNA 中的保守作用 生物学、生殖及其与癌症发病和进展相关的畸变，我们期待这项工作 与人类生物学有直接关系。