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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.
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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.
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.
开放通道毛细血管和毛细管抽水。
DOI: 10.1021/acs.langmuir.0c01360
发表时间: 2020-11-03
期刊: Langmuir : the ACS journal of surfaces and colloids
影响因子: --
作者: [Lee JJ, Berthier J, Kearney KE, Berthier E, Theberge AB]
通讯作者: Theberge AB
Localized Cell-Surface Sampling of a Secreted Factor Using Cell-Targeting Beads.
使用细胞靶向珠的分泌因子对局部细胞表面采样。
DOI: 10.1021/acs.analchem.0c02578
发表时间: 2020-10-20
期刊: Analytical chemistry
影响因子: 7.4
作者: [van Neel TL, Berry SB, Berthier E, Theberge AB]
通讯作者: Theberge AB
共 8 条
    Deciphering the role of chemical signals in inflammation with open microfluidic functional assays
    • 批准号:
      10456308
    • 项目类别:
    • 资助金额:
      $38.77万
    • 财政年份:
      2018
    • 负责人:
      Ashleigh Brooks Theberge
    • 依托单位:
    Deciphering the role of chemical signals in inflammation with open microfluidic functional assays
    • 批准号:
      10588933
    • 项目类别:
    • 资助金额:
      $1.37万
    • 财政年份:
      2018
    • 负责人:
      Ashleigh Brooks Theberge
    • 依托单位:
    Deciphering the role of chemical signals in inflammation with open microfluidic functional assays
    • 批准号:
      10556928
    • 项目类别:
    • 资助金额:
      $21.77万
    • 财政年份:
      2018
    • 负责人:
      Ashleigh Brooks Theberge
    • 依托单位:
    Deciphering the role of chemical signals in inflammation with open microfluidic functional assays
    • 批准号:
      10219302
    • 项目类别:
    • 资助金额:
      $39.21万
    • 财政年份:
      2018
    • 负责人:
      Ashleigh Brooks Theberge
    • 依托单位:
    海外基金