Optogenetic Control of Notch Signalling in vivo

体内Notch信号的光遗传学控制

• 批准号: RGPIN-2018-06781
• 负责人: Necakov, Aleksandar
• 金额: $ 2.99万
• 依托单位: Brock University
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2020
• 资助国家: 加拿大
• 起止时间: 2020-01-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=714441
• 关键词: Optogenetic; Control; Notch; Signalling; vivo

Our bodies are made up of trillions of tiny cells that are stuck together to make organs and tissues. To ensure that each cell maintains its unique identity, it is wrapped up in an outer sheet or 'plasma membrane' that compartmentalizes its contents. However, the cells in our bodies also need to communicate with one another in order to coordinate their function during development and homeostasis. The central theme of our research program is the Notch singling pathway. The Notch pathway is a cell-to-cell communication system used by multi-cellular animals to ensure the correct cell types form at a precise time and location in the body. Remarkably, this system is used over and over again during the formation of almost every cell type. Given the extensive requirement for Notch, it is not surprising that defects in this signaling pathway are associated with many human developmental diseases and cancers. In recent years, our understanding of the mechanisms that govern Notch signaling has grown significantly and provided insights into the cellular and genetic requirements for Notch. In particular, endocytic trafficking, the process by which cells internalize, sort, integrate, process, and secrete intercellular signaling components, has emerged as a critical regulator of Notch signaling. However, the complex, interconnected architecture of cellular signaling networks has posed formidable challenges to disentangling how cells process and integrate information. To circumvent these challenges, we apply a combination of genetic engineering, high resolution live imaging, and Optogenetics to precisely visualize and control components of the Notch pathway and the endocytic trafficking machinery with light. We use the fruit fly (Drosophila) and human cells as model systems for investigating the relationship between the signaling and trafficking machineries owing to the availability of sophisticated genetic tools, the high degree of conservation in the genes involved in Notch signaling and endocytic trafficking between flies and humans, and the ease with which students can gain the expertise required to perform sophisticated Optogenetic experiments. This approach will allow us to study the role of Notch signaling and endocytic trafficking in establishing boundaries between neighbouring tissues, in regulating stem cell differentiation, and will significantly advance our understanding of the mechanisms that control the Notch pathway and the endocytic trafficking machinery. We anticipate that our research program will generate both information-rich data sets and excellent opportunities for collaboration, will serve as an immersive training platform for undergraduate, graduate, and postgraduate students, and will provide fundamental mechanistic insights and novel hypotheses into the regulatory architecture of Notch signaling and endocytic trafficking during multicellular development.

我们的身体是由数以万亿计的微小细胞组成的，这些细胞粘在一起形成器官和组织。为了确保每个细胞保持其独特的身份，它被包裹在将其内容物隔开的外层薄片或“质膜”中。然而，我们体内的细胞也需要相互沟通，以协调它们在发育和动态平衡期间的功能。我们研究项目的中心主题是Notch单打路径。Notch途径是多细胞动物使用的一种细胞间通信系统，以确保在体内准确的时间和位置形成正确的细胞类型。值得注意的是，在几乎每种细胞类型的形成过程中，这个系统都会被反复使用。鉴于对Notch的广泛需求，该信号通路的缺陷与许多人类发育疾病和癌症有关也就不足为奇了。 近年来，我们对管理Notch信号的机制的了解有了显著的增长，并提供了对Notch的细胞和遗传要求的见解。特别是，细胞内化、分类、整合、加工和分泌细胞间信号成分的过程，已经成为Notch信号的关键调节因子。然而，复杂的、相互关联的细胞信令网络体系结构对细胞如何处理和整合信息提出了巨大的挑战。为了绕过这些挑战，我们应用基因工程、高分辨率实时成像和光遗传学的组合来精确地可视化和控制Notch途径的组件和用光进行内吞运输的机制。 我们使用果蝇(果蝇)和人类细胞作为模型系统来研究信号和运输机制之间的关系，这是由于复杂的基因工具的可用，涉及Notch信号和果蝇与人之间的胞内运输的基因高度保守，以及学生可以轻松获得执行复杂的光遗传学实验所需的专业知识。这种方法将使我们能够研究Notch信号和内吞运输在建立相邻组织之间的边界中的作用，在调节干细胞分化方面，并将极大地促进我们对控制Notch途径和内吞运输机制的机制的理解。我们预计，我们的研究计划将产生丰富的信息数据集和良好的合作机会，将作为本科生、研究生和研究生的沉浸式培训平台，并将为多细胞发育过程中Notch信号和内吞运输的调控架构提供基本的机械见解和新颖的假设。