Regulatory circuitry and mechanisms controlling cell fate in C. elegans

控制线虫细胞命运的调节电路和机制

• 批准号: 9908114
• 负责人: Iva S Greenwald
• 金额: $ 58.35万
• 依托单位: COLUMBIA UNIV NEW YORK MORNINGSIDE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-05-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9908114
• 关键词: Address; CRISPR/Cas technology; Caenorhabditis elegans; Cell Communication; Cells; Congenital Abnormality; Cues; Data Collection; Development; Developmental Biology; Diagnostic; Disease; Engineering; Epidermal Growth Factor Receptor; Fostering; Foundations; Gene Expression; Genetic; Gonadal structure; Health; Human; Lobular Neoplasia; Malignant Neoplasms; Medicine; Methods; Microfluidics; Molecular; Monitor; Pattern; Physiological; Population; Regulation; Signal Pathway; Signal Transduction; System; Technology; Therapeutic; Time; Vulva; Work; base; cell fate specification; gene function; genetic analysis; genetic approach; human disease; insight; life history; notch protein; personalized medicine; serial imaging; temporal measurement; tool

Project Summary/Abstract This proposal addresses how cells communicate with each other to control development by elucidating the signaling components, modulators and mechanisms that govern cell-cell interactions. Understanding the underlying genetic circuitry and molecular mechanisms is critical, because aberrant activity of these same signaling pathways have profound effects on human health, most notably as causal agents of cancer, congenital defects and diverse physiological disorders. Regulating signaling appropriately—in space or cell population, in time, strength or duration, or in combination with other signaling inputs—is thus crucial both for normal development and for insight into human disease. This proposal builds on foundational work on two key developmental paradigms in C. elegans: (i) specification of alternative cell fates by LIN- 12/Notch signaling in the gonad and (ii) the integration of LIN-12/Notch and EGFR-Ras-ERK signaling to pattern three distinct fates in the vulva. Although each paradigm has unique attributes, they provide a unified platform for elucidating regulatory circuitry and mechanisms underlying cell fate decisions and the function and regulation of major, conserved signaling systems. These C. elegans paradigms have a distinguished record of fostering discoveries that are directly applicable to basic human developmental biology and medicine, and a major reason is that they are especially amenable to genetic analysis. The proposed work will utilize a combination of classical genetic approaches and CRISPR/Cas9-engineering and other technologies to manipulate and monitor gene function, as well new microfluidic-based methods for longitudinal imaging and automated data collection that allows unprecedented cellular precision and temporal resolution in analyzing cell fate choice, gene expression, and signaling dynamics. The work will address three key gaps in understanding: (i) the relationship between signaling dynamics and cell fate specification, (ii) the regulatory circuitry and mechanisms that enable precise and robust spatial patterning, and (iii) the regulation of developmental progression through integrating life history and environmental cues with spatial patterning. The identification of mechanisms by which major, conserved signaling systems are regulated and integrated with each other is increasingly important in the era of personalized medicine and the deeper understanding of developmental mechanism we will achieve through these studies will be potentially applicable to developing diagnostic and therapeutic tools for human disease.

项目概要/摘要 该提案解决了细胞如何相互通信以通过以下方式控制发育： 阐明控制细胞间的信号传导成分、调节剂和机制 互动。了解潜在的遗传电路和分子机制至关重要， 因为这些相同信号通路的异常活动对人类具有深远的影响 健康，尤其是作为癌症、先天性缺陷和各种生理缺陷的致病因素 失调。适当调节信号传导——空间或细胞群、时间、强度或 因此，持续时间或与其他信号输入相结合对于正常情况至关重要 发展和洞察人类疾病。该提案建立在以下基础工作的基础上： 线虫的两个关键发育范例：（i）LIN-对替代细胞命运的规范 性腺中的 12/Notch 信号传导以及 (ii) LIN-12/Notch 和 EGFR-Ras-ERK 的整合 发出信号来塑造外阴三种不同的命运。尽管每个范式都有其独特之处 属性，它们提供了一个统一的平台来阐明监管电路和机制 潜在的细胞命运决定以及主要保守信号传导的功能和调节 系统。这些线虫范式在促进发现方面有着杰出的记录： 直接适用于基础人类发育生物学和医学，一个主要原因 是它们特别适合基因分析。拟议的工作将利用 经典遗传方法与 CRISPR/Cas9 工程等的结合 操纵和监测基因功能的技术，以及基于微流体的新方法 用于纵向成像和自动数据收集，允许前所未有的细胞 分析细胞命运选择、基因表达和信号传导的精度和时间分辨率 动力学。这项工作将解决理解上的三个关键差距：(i) 信号动力学和细胞命运规范，（ii）调节电路和机制 实现精确而稳健的空间模式，以及（iii）发育进程的调节 通过将生活史和环境线索与空间图案相结合。鉴定 主要的保守信号系统的调节和整合机制 在个性化医疗和更深层次的时代，彼此之间的关系变得越来越重要。 通过这些研究我们将获得对发展机制的理解 可能适用于开发人类疾病的诊断和治疗工具。