CAREER: microRNA-mediated regulation of dosage sensitive genes involved in morphogenesis

职业：微小RNA介导的形态发生剂量敏感基因的调节

基本信息

批准号：
2238425
负责人：
Luisa Cochella
金额：
$ 93.5万
依托单位：
Johns Hopkins University
依托单位国家：
美国
项目类别：
Continuing Grant
财政年份：
2023
资助国家：
美国
起止时间：
2023-04-01 至 2028-03-31
项目状态：
未结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=2238425&HistoricalAwards=false
关键词：
CAREER microRNA mediated regulation dosage

项目摘要

The human genome has about 20,000 genes, which contain the information to produce proteins that carry out a myriad of functions in and around the cells that make up our bodies. For the correct development and function of human bodies, and those of any organism, specific proteins need to be produced in specific tissues or organs, at specific times. It is also essential that proteins be produced at their correct levels to achieve their normal cellular functions. A number of genes called “dosage sensitive genes” produce proteins that are exquisitely sensitive to changes in level, meaning that too much or too little of these proteins is detrimental to the cells, and thus to the living organism. Changes in the level of these proteins, which may be caused by having too few or too many copies of the genes that produce them, often result in human disease. This project studies a mechanism for ensuring that genes are not produced at overly high levels. The researchers study this type of regulation in a model organism that enables a quantitative and detailed examination of development and of protein production under different conditions. They use a microscopic nematode, C. elegans, in which they can modify genes and measure the output during development. Studying such mechanisms can inform our understanding of why some gene mutations cause disease, and could ultimately teach us how to manipulate gene dosage for therapeutic purposes. A significant fraction of genes in a genome are dosage sensitive, such that copy number variation (loss or gain of even a single genomic copy) often results in disease. Identifying the regulators that enable precise gene expression levels is critical to fully understand gene regulation and to reveal modifiers of disease relevant genes. MicroRNAs (miRNAs) are quantitative repressors of target mRNAs at the post-transcriptional level and it has been proposed that dosage sensitive genes rely on miRNA-mediated regulation. The researchers have established an experimental paradigm to test this in the context of a key developmental process that relies on precise gene dosage: animal morphogenesis. miR-100 is the most deeply conserved animal miRNA, present across all Eumetazoa. Despite its conservation, the molecular, cellular and organismal roles of miR-100 are practically unknown. The researchers propose to use C. elegans to study the function of this ancient animal miRNA and gain insight into the regulation of dosage sensitive genes in development. They have evidence that ECM components are conserved targets of miR-100 in worms, fish, mice and humans, suggesting that this miRNA may be involved in cellular signaling and adhesion in C. elegans, and likely other animals. Using genetic approaches, the researchers will reveal the role of this ancient miRNA in C. elegans, provide insight into its function in other animals based on shared targets, define the role of a dosage dependent regulator of morphogenesis, and gain new understanding of the contribution of miRNAs as regulators of dosage sensitive genes.This award is co-funded by the Developmental Systems Program in the Integrative Organismal Systems Division and the Genetic Mechanisms Program in the Molecular and Cellular Biosciences Division of the Directorate for Biological Sciences.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

人类基因组有大约20,000个基因，其中包含可产生蛋白质的信息，这些蛋白质在构成我们身体的细胞内及其周围进行了无数功能。为了使人体的正确发育和功能以及任何生物体的特定蛋白质，需要在特定的组织或器官中产生特定的蛋白质。也必须以正确的水平生产蛋白质以达到其正常的细胞功能。许多称为“剂量敏感基因”的基因会产生对水平变化完全敏感的蛋白质，这意味着这些蛋白质中的太多或太少对细胞有害，因此对生命组织有害。这些蛋白质水平的变化可能是由于产生它们的基因的副本太少或太多而引起的，通常会导致人类疾病。该项目研究了一种确保基因在过高水平过高产生的机制。研究人员在模型生物体中研究了这种调节，该调节能够对不同条件下的发育和蛋白质产生进行定量和详细的研究。他们使用微型线虫，秀丽隐杆线虫，其中可以修改基因并测量发育过程中的输出。研究这种机制可以告知我们对某些基因突变引起疾病的理解，并最终可以教会我们如何以治疗目的操纵基因剂量。基因组中的很大一部分基因对剂量敏感，因此拷贝数变化（甚至单个基因组拷贝的损失或增益）通常会导致疾病。识别能够使精确基因表达水平的调节剂对于完全了解基因调节并揭示疾病相关基因的修饰剂至关重要。 microRNA（miRNA）是在转录后水平上靶标mRNA的定量代表，并且已经提出剂量敏感基因依赖于miRNA介导的调节。研究人员已经建立了一个实验范式，以在依赖精确基因剂量的关键发育过程的背景下进行测试：动物形态发生。 mir-100是所有Eumetazoa中最深处的动物miRNA。尽管保存了miR-100的分子，细胞和有机作用，但实际上尚不清楚。研究人员已经建立了一个实验范式，以在依赖精确基因剂量的关键发育过程的背景下进行测试：动物形态发生。 mir-100是所有Eumetazoa中最深处的动物miRNA。尽管保存了miR-100的分子，细胞和有机作用，但实际上尚不清楚。研究人员建议使用秀丽隐杆线虫研究这种古老的动物miRNA的功能，并深入了解发育中剂量敏感基因的调节。他们有证据表明，ECM成分是蠕虫，鱼类，小鼠和人类中miR-100的保守靶标，表明该miRNA可能参与了秀丽隐杆线虫以及可能的其他动物中的细胞信号传导和粘合剂。使用遗传方法，研究人员将揭示这种古老的miRNA在秀丽隐杆线虫中的作用，从共同的目标基于共同的目标中对其在其他动物中的功能进行洞察力，定义了依赖剂量的形态发生调节因子的作用，并获得了对miRNA作为剂量敏感基因的调节器的贡献的新理解。该奖项反映了NSF的法定任务，并通过评估使用该基金会的知识分子优点和更广泛的影响标准，反映了NSF的法定任务。