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: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=744801
• 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.

我们的身体是由数万亿个微小的细胞组成的，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，它被包裹在一个外层或“质膜”中，将其内容物分隔开。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体内平衡过程中的功能。我们研究计划的中心主题是Notch singling pathway。Notch通路是多细胞动物使用的细胞间通讯系统，以确保在体内的精确时间和位置形成正确的细胞类型。值得注意的是，这个系统在几乎每种细胞类型的形成过程中都被反复使用。由于Notch的广泛需求，这一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Notch信号转导机制的理解显著增长，并为Notch的细胞和遗传需求提供了见解。特别地，内吞运输（细胞内化、分选、整合、加工和分泌细胞间信号传导组分的过程）已经成为Notch信号传导的关键调节剂。然而，细胞信号网络的复杂、互连的架构对解开细胞如何处理和整合信息提出了巨大的挑战。为了规避这些挑战，我们将基因工程、高分辨率实时成像和光遗传学相结合，利用光精确地可视化和控制Notch途径和内吞运输机制的组成部分。我们使用果蝇（果蝇）和人类细胞作为模型系统，用于调查信号和贩运机制之间的关系，由于复杂的遗传工具的可用性，高度保守的Notch信号和内吞运输的基因在苍蝇和人类之间，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信号传导和内吞运输在建立相邻组织之间的边界，调节干细胞分化中的作用，并将显着推进我们对控制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信息丰富的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，并将提供基本的机制见解和新的假设到调控架构Notch信号和内吞贩运在多细胞发育。