Molecular regulation of germline development

种系发育的分子调控

• 批准号: 9797218
• 负责人: JUDITH KIMBLE
• 金额: $ 55.47万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-15 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9797218
• 关键词: Address; Animals; Atrophic; Binding; Binding Proteins; Biological; Biological Process; Caenorhabditis elegans; Clinical; Cues; Defect; Development; Diagnosis; Disease; Epilepsy; Equilibrium; Family; Fertility; Gender; Germ Cells; Glean; Goals; Health; Homeostasis; Human; Infertility; Intervention; Learning; Link; Logic; Malignant Neoplasms; Mammals; Messenger RNA; Methods; Molecular; Nematoda; Neurobiology; Organism; Pattern; Phylogeny; Physiological; Proteins; Proteome; RNA; RNA-Binding Proteins; Regulation; Repression; Research; Role; Signal Transduction; Starvation; Stem cells; Temperature; Time; Tissues; Totipotency; Translations; Work; combinatorial; genetic regulatory protein; genome-wide; human disease; in vivo; insight; multidisciplinary; novel therapeutics; self-renewal; therapy design; transcriptome

PROJECT SUMMARY/ABSTRACT Our goal is to understand molecular principles of germ cell regulation. We focus on a decision that all stem cells make: to self-renew or begin differentiation. This decision is fundamental in all tissues and is critical to human health. Regulators that promote the stem cell state must be active to drive self-renewal and totipotency, but must be inactivated for differentiation; conversely, regulators that are critical for differentiation must be inactive in stem cells, but then become activated at the right time and place for differentiation. Regulation of the transition – from self-renewal to differentiation – balances the two states and is fundamental to development, homeostasis and human health. Defects in that balance cause human disease, including tissue atrophy and cancer. RNA regulation is a major conserved theme in germ cells from worms to mammals. Key RNA-binding proteins regulate self-renewal and differentiation across animal phylogeny. We focus on PUF (Pumilio and FBF) RNA- binding proteins, which have pivotal roles in germ cells and neurobiology across animal species. In humans, defective PUF proteins cause infertility and epilepsy. PUF proteins have many features in common: They sculpt the transcriptome and proteome by binding to more than 1000 mRNAs, with 100's conserved between nematodes and humans; they regulate target mRNAs by diverse mechanisms, including activation and repression; and they work in a combinatorial fashion with partner proteins, most of which are also conserved. The challenge now is to understand how PUF proteins regulate the balance between self-renewal and differentiation in molecular terms and in a native in vivo context. Our proposal addresses this challenge with a sharp focus on four issues. First, we elucidate how distinct PUF mechanisms – activation and repression – work together to control germ cell fates. Second, we elucidate how PUF partnerships operate in vivo, to understand their logic and glean general principles. Third, we elucidate regulation of the balance between self-renewal to differentiation. Fourth, we elucidate how a functional module of multiple PUF proteins and partners maintains stem cells under diverse physiological and environmental conditions. These four issues are fundamental to all PUF regulation. For each, we take a multidisciplinary and multiscale approach and exploit an exceptionally tractable network. Together, our findings will advance our understanding germ cell regulation with potential for broad impact, including of new therapy designs for humans.

项目摘要/摘要 我们的目标是了解生殖细胞调控的分子原理。我们专注于一个决定，所有这些都源于 细胞制造：自我更新或开始分化。这一决定在所有组织中都是基本的，并且对 人类健康。促进干细胞状态的监管机构必须积极推动自我更新和全能， 但必须停用以实现差异化；相反，对差异化至关重要的调节器必须 在干细胞中不活跃，但随后在适当的时间和地点被激活以进行分化。对 从自我更新到差异化的过渡平衡了这两个国家，是发展的基础， 动态平衡与人类健康。这种平衡的缺陷会导致人类疾病，包括组织萎缩和 癌症。 RNA调控是从蠕虫到哺乳动物的生殖细胞中一个主要的保守主题。关键的RNA结合蛋白 调节动物进化过程中的自我更新和分化。我们专注于PUF(Pumilio和FBF)RNA- 结合蛋白，在动物物种的生殖细胞和神经生物学中具有关键作用。在人类身上， PUF蛋白缺陷会导致不孕和癫痫。PUF蛋白有许多共同的特征：它们塑造 转录组和蛋白质组通过与1000多个mRNAs结合，与100‘端的S保守 线虫和人类；它们通过不同的机制调节靶mRNAs，包括激活和 抑制；它们与伙伴蛋白以组合的方式工作，其中大多数也是保守的。 现在的挑战是了解PUF蛋白如何调节自我更新和 在分子方面和在体内自然环境中的差异。我们的建议通过以下方式解决这一挑战 突出抓好四个问题。首先，我们阐明了不同的PUF机制--激活和抑制--是如何工作的 共同控制生殖细胞的命运。其次，我们阐明PUF伙伴关系在体内是如何运作的，以了解 他们的逻辑和收集的一般原则。第三，我们阐明了自我更新与自我更新之间的平衡调节 差异化。第四，我们阐明了多个PUF蛋白和伙伴的功能模块是如何维持的 干细胞在不同的生理和环境条件下。这四个问题是所有问题的基础 PUF法规。对于每个人，我们都采取了多学科和多规模的方法，并开发了一个例外的 易于管理的网络。总之，我们的发现将促进我们对生殖细胞调节的理解，具有潜在的 广泛的影响，包括对人类的新疗法设计。