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

• 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
• 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

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.

抽象的。合成生物学结合了生物学和工程原理以调节细胞 过程，并且正在成为生物医学研究的重要领域。迄今为止，合成生物学具有 主要集中于操纵细胞内部的过程，最著名的是控制基因表达或 代谢，并将其排列到执行离散功能的模块中。相比之下，电流细胞 - 基于培养的合成方法不适合操纵控制相互作用的过程 在细胞之间产生预期的结果，在组织水平上，这种能力特别有价值 在组织工程和再生医学领域。需要的是在遗传上可进行的 可以快速创建，测试，优化和 在部署并进一步精炼的系统中，多样化 生物技术应用。我们建议通过建立果蝇系统来满足这一要求 用于设计控制组织行为的合成细胞间信号。我们已经开发了原型 关于Notch激活的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 受体的域。我们已经将合成受体与一种用于控制的新遗传方案配对 配体/受体相互作用，进行功能性筛选并改变合成受体输出。第一的， 我们将通过进行新的大规模屏幕来使我们的合成受体曲目多样化 异源力敏感的切割结构域。其次，我们将严格评估我们的新受体 它们适合在电路中使用并表征其重要响应参数，例如 信号的效力和调节能力。最后，我们将组装合成信号的电路 模块在上皮组织内产生可预测的输出。如果成功，提议 实验将是朝着我们建立可拖动体内系统的长期目标迈出的重要一步 用于开发细胞电池信号技术，该技术在组织工程和 再生医学。