Developing a platform for engineering customizable cell-cell signaling in vivo

开发用于设计可定制的体内细胞间信号传导的平台

基本信息

批准号：
10528148
负责人：
Paul Langridge
金额：
$ 22.2万
依托单位：
AUGUSTA UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-01 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10528148
关键词：
Area Assessment tool Behavior Biology Biomedical Engineering Biomedical Research Biotechnology Cell Communication Cell Culture System Cell Culture Techniques Cell physiology Cells Characteristics Collection Communities Complex Congenital Abnormality Cues Custom Development Disease Drosophila genus Endocytosis Engineering Environment Epithelial Future Gene Expression Gene Expression Profile Generations Genes Genetic Genetic Screening Goals Human Development Ligands Limb structure Link Mammalian Cell Metabolism Mission National Institute of Child Health and Human Development Natural regeneration Outcome Output Pattern Peptide Hydrolases Physiology Process Protocols documentation Public Health Regenerative Medicine Regulation Research Series Signal Transduction Societies System T-Lymphocyte Technology Testing Therapeutic TimeLine Tissue Engineering Tissues Transcriptional Regulation United States National Institutes of Health Wing Work base cell behavior design experimental study extracellular force sensor in vivo infancy intercellular communication interest mechanical force new technology notch protein predicting response prototype receptor response synthetic biology tissue repair tool wound healing

项目摘要

Abstract. Synthetic biology combines biological and engineering principles to regulate cellular processes, and is emerging as an important area of biomedical research. To date, synthetic biology has focused largely on manipulating processes inside cells, most notably to control gene expression or metabolism, and arranging them into modules that perform discrete functions. By contrast, current cell- culture based synthetic approaches are ill-equipped to manipulate processes that control interactions between cells to create desired outcomes at the tissue level, a capacity that would be of particular value in the fields of tissue engineering and regenerative medicine. What is needed is a genetically tractable in vivo platform within which synthetic cell-cell signaling tools can be rapidly created, tested, optimized and diversified, before they are deployed and further refined in systems that have therapeutic and biotechnological applications. We propose to fulfill this requirement by establishing a Drosophila system for designing synthetic intercellular signaling that controls tissue behavior. We have developed prototype synthetic ligand/receptor systems predicated on the basic mechanism of Notch activation, where mechanical force exerted by ligand endocytosis induces the cleavage of an extracellular “force-sensitive” domain of the receptor. We have paired our synthetic receptors with a new genetic protocol for controlling ligand/receptor interactions, conducting functional screens and altering synthetic receptor outputs. First, we will diversify our repertoire of synthetic receptors by conducting a large-scale screen for new heterologous force-sensitive cleavage domains. Second, we will rigorously assess our new receptors for their suitability to be used in circuits and characterize their important response parameters, such as the potency of signal and capacity for regulation. Last, we will assemble circuits of synthetic signaling modules to produce predictable outputs within an epithelial tissue. If successful, the proposed experiments will be a significant step toward our long-term goal of establishing a tractable in vivo system for developing cell-cell signaling technology that has future applications in tissue engineering and regenerative medicine.

摘要：合成生物学结合了生物学和工程学原理来调节细胞。迄今为止，合成生物学已成为生物医学研究的一个重要领域。主要关注细胞内部的操纵过程，最显着的是控制基因表达或新陈代谢，并将它们排列成执行离散功能的模块。基于文化的合成方法不足以操纵控制相互作用的过程细胞之间在组织水平上创造期望的结果，这种能力具有特殊的价值在组织工程和再生医学领域，我们需要的是一种基因上易于处理的细胞。体内平台，可以在其中快速创建、测试、优化和合成合成细胞信号工具在将它们部署到具有治疗和治疗作用的系统中并进一步完善之前，应进行多样化的处理我们建议通过建立果蝇系统来满足这一要求。为了设计控制组织行为的合成细胞间信号传导，我们开发了原型。基于Notch激活基本机制的合成配体/受体系统，其中配体内吞作用施加的机械力诱导细胞外“力敏感”的裂解我们已经将我们的合成受体与新的控制基因协议配对。配体/受体相互作用，进行功能筛选并改变合成受体输出。我们将通过大规模筛选新的受体来使我们的合成受体库多样化其次，我们将严格评估我们的新受体。它们是否适合在电路中使用并表征其重要的响应参数，例如最后，我们将组装合成信号电路。如果成功，建议的模块在上皮组织内产生可预测的输出。实验将是我们朝着建立易于处理的体内系统的长期目标迈出的重要一步开发细胞信号传导技术，该技术未来可应用于组织工程和再生医学。