Resolving the spatial dynamics of single cell circadian regulatory networks

项目摘要/摘要 生理和代谢健康依赖于生物过程的昼夜节律与 环境。自从18世纪第一次利用植物系统对昼夜节律进行有记录的研究以来， 模型生物对于定义振荡器的转录机制和揭示 时钟对健身的重要性。这项工作的大部分是基于整个生物体或器官水平的研究 许多关于细胞特异性基因调控如何导致细胞时钟和 协调一致的生理反应。我们知道基因调控网络在模拟基因方面是有效的。 表达动态，但解析具有时间分辨率的细胞类型特定网络仍然是一个重要的 挑战。 为了描绘跨细胞类型的调节连接，需要单个细胞视图来开发网络 反映真实细胞状态的模型，而不是细胞或组织组之间的差异。只有这些 细胞特异性网络我们可以开始开发可测试的假说，以了解调控变异的基础是什么 生理反应。这项建议描述了一种利用遗传和分子 植物模型系统拟南芥和最新单细胞技术的力量1)识别 具有不同振荡器的细胞类型及其潜在的调控网络，2)剖析细胞类型特异性 昼夜节律网络维持同步的生理反应，以及3)扰动细胞类型的特异性 昼夜节律基因调控网络，并评估其生理后果。 在接下来的五年里，我们的目标是描绘出整个单细胞昼夜节律基因调控网络 具有空间分辨率的整个器官。我们的拟南芥模型系统将用来剖析不同的细胞 特定类型的昼夜节律控制生理反应。通过对组织特异性的扰动 基因调控网络我们将更好地理解细胞基因编程是如何协调的。 我们的长期目标是将拟南芥发展成一个模型，以了解细胞特异性基因调控 网络影响组织间的通信。这项研究将对我们如何解释产生有价值的见解 昼夜节律基因调控变异对人类健康的影响及靶向治疗的应用。

Project Summary/Abstract Physiological and metabolic health relies on the circadian alignment of biological processes with the environment. Since the first documented study of circadian rhythms in the 18th century using a plant system, model organisms have been critical for defining the transcriptional mechanism of the oscillator and revealing the importance of the clock on fitness. Most of this work is based on whole organism or organ level studies leaving many mechanistic questions about how cell specific gene regulation leads to coordination of cellular clocks and a concerted physiological response. We know that gene regulatory networks are effective at modeling gene expression dynamics but resolving cell-type specific networks with time resolution remains a significant challenge. To delineate regulatory connections across cell-types, a single cell view is needed to develop network models that reflect the true cell state rather than the variation among groups of cells or tissues. Only with these cell specific networks can we begin to develop testable hypotheses about what regulatory variation underlies physiological responses. This proposal describes a research strategy that leverages the genetic and molecular power of the plant model system Arabidopsis thaliana and the latest single cell technologies to 1) Identify the cell-types with distinct oscillators and their underlying regulatory networks, 2) Dissect how cell-type specific circadian regulatory networks maintain a synchronized physiological response, and 3) Perturb cell-type specific circadian gene regulatory networks and evaluate their physiological consequences. Over the next five years, our goals are to delineate single cell circadian gene regulatory networks across an entire organ with spatial resolution. Our Arabidopsis model system will be used to dissect how distinct cell type specific circadian regulation controls a physiological response. Through perturbations to tissue specific gene regulatory networks we will gain a better understanding of how cellular gene programming is coordinated. Our long-term goal is to develop Arabidopsis into a model for understanding how cell specific gene regulatory networks influence inter-tissue communication. This research will generate valuable insight into how we interpret the influence of circadian gene regulatory variation on human health and the application of targeted therapies.