Regulatory circuitry and mechanisms controlling cell fate in C. elegans

项目总结/摘要 该提案解决了细胞如何相互沟通，以控制发展， 阐明了控制细胞-细胞的信号成分、调节剂和机制， 交互.了解潜在的遗传电路和分子机制是至关重要的， 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，特别是作为癌症、先天性缺陷和各种生理缺陷的致病因素， 紊乱适当地调节信号-在空间或细胞群体中，在时间，强度或 持续时间，或与其他信号输入相结合，因此对于正常的 发展和洞察人类疾病。本提案基于以下方面的基础性工作： C.（i）通过LIN-1000对替代细胞命运的说明。 （ii）LIN-12/Notch和EGFR-Ras-ERK的整合 发出信号在外阴形成三种不同的命运虽然每种范式都有其独特的 属性，它们提供了一个统一的平台，阐明调控电路和机制 潜在的细胞命运决定以及主要保守信号的功能和调节 系统.这些C.优雅的范例有一个杰出的记录， 直接适用于基本的人类发育生物学和医学， 是因为它们特别适合基因分析拟议的工作将利用一个 经典遗传方法和CRISPR/Cas9工程和其他方法的组合 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， 分析细胞命运选择、基因表达和信号传导的精确度和时间分辨率 动力学这项工作将解决理解方面的三个关键差距： 信号动力学和细胞命运的规范，（ii）调控电路和机制， 使精确和强大的空间模式，和（iii）发展进程的调节 通过整合生活史和环境线索与空间模式。识别 主要的，保守的信号系统被调节和整合的机制， 在个性化医疗时代，彼此越来越重要， 我们将通过这些研究来理解发展机制， 可能适用于开发人类疾病的诊断和治疗工具。

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.