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

基本信息

批准号：
10621092
负责人：
Ashleigh Brooks Theberge
金额：
$ 41.99万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-08-01 至 2028-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10621092
关键词：
Address Affect Area B-Lymphocytes Bacteria Biological Assay Biological Process Biomedical Engineering Biomimetics Blood capillaries Cells Chemical Stimulation Chemicals Communication Complex Development Disease Ecosystem Engineering Fiber Fibroblasts Future Health Home Human Immune Immune response Inflammation Inflammatory Investigation Life Mass Fragmentography Mediating Methods Microbe Microfluidics Molecular Organ Paracrine Communication Phase Positioning Attribute Production Role Sampling Signal Transduction Signaling Molecule Signaling Protein Solid System Time Translating Vocabulary Volatilization Work cell type commensal bacteria commensal microbes cost effective eosinophil experimental study fungus human disease innovation microbial microbial community monocyte neutrophil novel novel strategies paracrine pathogen pathogenic bacteria sample collection small molecule therapy development tool user-friendly volatile organic compound

项目摘要

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. 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 or when different cell types are present in the microenvironment? 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 cell signaling, (2) specialized culture platforms for complex human-bacteria-fungal multikingdom culture, (3) integration of microbial co- and multiculture systems with volatile organic compound (VOC) sampling to study how volatiles mediate microbial dialogue, and (4) at-home biofluid sampling and stabilization platforms to probe the human immune response over time. The present proposal expands our lab’s capabilities in areas (1) and (3), with the possibility to extend to (2) and (4) in future work. This proposal will probe unanswered questions in two areas of human cell signaling: (i) paracrine signaling between eosinophils and fibroblasts and (ii) paracrine/physical contact-mediated signaling between neutrophils, monocytes, and B cells. Further, we will develop novel culture platforms that enable microbial and multikingdom (e.g., bacteria, fungi) VOC communication and integrated sampling for gas chromatography-mass spectrometry (GC-MS) analysis. This culture system will, for the first time, enable controlled spatial positioning of multiple microbial cultures, a user-friendly setup that can be operated with simple pipettes, fluidic channels to deliver media and chemical stimulation, and integration of solid phase micoextraction (SPME) fibers for VOC sample collection. Central to this proposal is the use of ‘open’ microfluidics and spontaneous capillary flow. We are leaders in open microfluidics and have a strong track record of developing user-friendly, cost-effective methods to perform microscale multiculture experiments within standard well plates and cultureware familiar to biologists. The proposed work builds on our capabilities and embraces significant engineering challenges in doing triculture with sensitive primary cells and innovating an entirely new approach for study VOCs in microbial cultures. 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; they will also enable study of microbial communities that are essential to maintaining human health (commensal microbes) and those that lead to disease (pathogens). Further, the bioengineering and microfluidic approaches developed will translate to other biomimetic culture platforms and fundamental signaling investigations.

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。解剖细胞类型之间的分子对话具有挑战性，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？生物功能如何当混合物中存在分子或存在不同的细胞类型时，不同微环境？微生物（例如我们体内存在的细菌和真菌）如何影响分子景观？我们的实验室正在开发新工具来探究这些问题，包括（1）微观共同和多元培养方法可以使细胞类型精确定位以研究细胞信号，（2）专业培养物复杂的人类 - 细菌多数字培养的平台，（3）微生物共同和具有挥发性有机化合物（VOC）采样的多元文化系统，以研究挥发性介质微生物如何对话和（4）在家中生物流体采样和稳定平台，以探测人类免疫反应随着时间的推移。本提案扩大了我们实验室在（1）和（3）领域的能力，有可能扩展到以后的工作（2）和（4）。该提案将在人类细胞信号的两个领域探讨未解决的问题：（i）嗜酸性粒细胞和成纤维细胞之间的旁分泌信号传导以及（ii）旁分泌/物理接触介导的信号传导在中性粒细胞，单核细胞和B细胞之间。此外，我们将开发新颖的文化平台微生物和多级（例如细菌，真菌）VOC通信和气体集成抽样色谱 - 质谱法（GC-MS）分析。这种文化系统将首次启用多种微生物培养物的控制空间定位，这是一种用户友好的设置，可以简单地操作移液器，流体通道，以传递培养基和化学刺激以及固相的整合（SPME）用于VOC样品收集的纤维。该建议的核心是使用“开放”微流体和赞助毛细血管流。我们是开放式微流体学领导者，并且有良好的发展记录用户友好，具有成本效益的方法，用于在标准井板中进行微观多元文化实验和生物学家熟悉的培养软件。拟议的工作以我们的能力为基础，并具有重要的用敏感的原代细胞和创新的全新方法进行工程挑战用于微生物培养的研究。提出的方法将增强对信号的理解参与有害的长期感染，这对于许多众多疗法的发展至关重要炎症条件；他们还将启用对维持的微生物社区的研究人类健康（共生微生物）和导致疾病的人（病原体）。此外，生物工程开发的微流体方法将转化为其他仿生文化平台和基本信号调查。

项目成果

期刊论文数量（12）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Miniaturizing chemistry and biology using droplets in open systems.

DOI：
10.1038/s41570-023-00483-0
发表时间：
2023-06
期刊：
NATURE REVIEWS CHEMISTRY
影响因子：
36.3
作者：
Zeng, Yuting;Khor, Jian Wei;van Neel, Tammi L.;Tu, Wan-chen;Berthier, Jean;Thongpang, Sanitta;Berthier, Erwin;Theberge, Ashleigh B.
通讯作者：
Theberge, Ashleigh B.

At-Home Saliva Sampling in Healthy Adults Using CandyCollect, a Lollipop-Inspired Device.

DOI：
10.1021/acs.analchem.3c00462
发表时间：
2023-07-11
期刊：
ANALYTICAL CHEMISTRY
影响因子：
7.4
作者：
Tu, Wan-chen;McManamen, Anika M.;Su, Xiaojing;Jeacopello, Ingrid;Takezawa, Meg G.;Hieber, Damielle L.;Hassan, Grant W.;Lee, Ulri N.;Anana, Eden V.;Locknane, Mason P.;Stephenson, Molly W.;Shinkawa, Victoria A. M.;Wald, Ellen R.;DeMuri, Gregory P.;Adams, Karen N.;Berthier, Erwin;Thongpang, Sanitta;Theberge, Ashleigh B.
通讯作者：
Theberge, Ashleigh B.

Open-Channel Capillary Trees and Capillary Pumping.