Deciphering the role of chemical signals in inflammation with open microfluidic functional assays

通过开放微流控功能分析解读化学信号在炎症中的作用

• 批准号: 9751911
• 负责人: Ashleigh Brooks Theberge
• 金额: $ 45.95万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9751911
• 关键词: Address; Affect; Anti-inflammatory; Area; Asthma; Bacteria; Behavioral Assay; Benign; Biological; Biological Assay; Biological Process; Biomedical Engineering; Biomimetics; Blood Vessels; Blood capillaries; Cell physiology; Cells; Chemicals; Chronic; Communication; Complex; Contracts; Development; Dimensions; Disease; Engineering; Environmental Exposure; Fatty Acids; Fibroblasts; Fibrosis; Gel; Human; Hyperplasia; Immune; Immune response; Infection; Inflammation; Inflammatory; Malignant Neoplasms; Mass Spectrum Analysis; Methods; Microbe; Microfluidics; Molecular; Mucous body substance; Physiology; Play; Positioning Attribute; Production; Resolution; Role; Signal Transduction; Signaling Molecule; Signaling Protein; Structure; Time; Translating; Vasodilation; Vocabulary; Work; angiogenesis; base; cell type; cost effective; experimental study; fungus; human disease; innovation; metabolomics; novel; response; small molecule; therapy development; tool; user-friendly

Project Abstract Small molecule and protein signals provide a rich vocabulary for cellular communication. The production and consequences of these signals are exquisitely sensitive to cellular context and microenvironment. For example, synthesis of pro-inflammatory, anti-inflammatory, and pro-resolution oxylipins in response to environmental exposures or wounding is tightly controlled by immune cells that can shift oxylipin production on the minute timescale as the immune response progresses in real time. Furthermore, the same signaling molecule can bring about an entirely different downstream biological response depending on microenvironmental context. Dissecting the molecular dialogue between cell types is challenging, and new methods are required to address fundamental questions: What is the downstream biological function of each signaling molecule? How is the biological function different when molecules are present in mixtures? How do microbes – like the bacteria and fungi present in our bodies – affect the molecular landscape? Our lab is developing new tools to probe these questions including (1) microscale co- and multiculture methods that enable precise positioning of cell types to study signaling, (2) integration of microculture with small molecule extraction methods for downstream metabolomics analysis using mass spectrometry, (3) specialized culture platforms and extraction methods to isolate signals from complex human-bacteria-fungal multikingdom culture, and (4) novel cell-based behavioral assays to probe the effects of chemical signals on biological function (including angiogenesis, mucus production, and immune cell function). The present proposal expands our lab’s capabilities in areas (1) and (4). This proposal will create innovative functional assays to study vasodilation (blood vessel expansion, a hallmark of inflammation) and fibroblast myodifferentiation (which leads to harmful fibrosis and remodeling in chronic inflammation). Central to this proposal is the use of ‘open’ microfluidics and spontaneous capillary flow to sculpt gel structures in three dimensions with sub-millimeter precision. Our lab has made significant advances in directing gel flows using open microfluidics, resulting in user-friendly, cost-effective methods to perform microscale multiculture experiments within standard well plates. The proposed work builds on our capabilities and embraces a significant engineering challenge: producing blood vessel mimics that can dynamically dilate and contract while being easy to multiplex in order to study large sets of signaling molecules. The proposed methods will enhance the understanding of the signals involved in detrimental prolonged inflammation, critical to the development of better therapies for numerous inflammatory conditions. Further, the bioengineering and microfluidic approaches developed will translate to other three dimensional biomimetic culture platforms.

项目摘要 小分子和蛋白质信号为细胞通信提供了丰富的词汇。生产和 这些信号的后果对细胞上下文和微环境完全敏感。例如， 对环境的促炎，抗炎和促分辨率的黄磷脂的合成 暴露或获胜由免疫细胞严格控制，这些细胞可以在一分钟内转移奥甲ipin的产生 随着免疫冲源的实时进行时间尺度。此外，相同的信号分子可以带来 根据微环境环境，下游生物学反应完全不同。 剖析细胞类型之间的分子对话是挑战，需要新的方法来解决 基本问题：每个信号分子的下游生物学功能是什么？怎么样 当混合物中存在分子时，生物学功能不同？微生物如何像细菌和 我们体内存在的真菌 - 影响分子景观？我们的实验室正在开发新工具来探究这些 包括（1）微观共同和多元文化方法在内的问题，可以将细胞类型精确定位到 研究信号传导，（2）将微培养与小分子提取方法整合到下游 使用质谱，（3）专业培养平台和提取方法的代谢组学分析 来自复杂的人类 - 肉杆菌多菌培养物的分离信号，以及（4）基于细胞的新型行为 探测化学信号对生物学功能的影响的测定（包括血管生成，粘液产生， 和免疫细胞功能）。本提案扩大了我们实验室在（1）和（4）领域的能力。这个建议 将创建创新的功能测定来研究血管舒张（血管扩张，是 炎症）和成纤维细胞肌相分化（这导致有害纤维化和重塑 炎）。该提案的核心是使用“开放”微流体和赞助毛细管流到雕刻 具有亚毫米精度的三个维度的凝胶结构。我们的实验室在 使用开放的微流体引导凝胶流，从而导致用户友好，具有成本效益的方法 标准井板中的微观多元文化实验。拟议的工作以我们的能力为基础 并包含重大的工程挑战：产生可以动态扩张的血管模仿 和收缩，同时易于多重，以研究大量信号分子。提议 方法将增强对有害长期感染涉及的信号的理解，对 为众多炎症条件开发更好的疗法。此外，生物工程和 开发的微流体方法将转化为其他三维仿生培养平台。