# Deciphering the role of chemical signals in inflammation with open microfluidic functional assays - Bouker Diversity Supplement

> **NIH NIH R35** · UNIVERSITY OF WASHINGTON · 2024 · $48,115

## Abstract

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 t...

## Key facts

- **NIH application ID:** 11013272
- **Project number:** 3R35GM128648-07S1
- **Recipient organization:** UNIVERSITY OF WASHINGTON
- **Principal Investigator:** Ashleigh Brooks Theberge
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $48,115
- **Award type:** 3
- **Project period:** 2018-08-01 → 2028-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/11013272

## Citation

> US National Institutes of Health, RePORTER application 11013272, Deciphering the role of chemical signals in inflammation with open microfluidic functional assays - Bouker Diversity Supplement (3R35GM128648-07S1). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/11013272. Licensed CC0.

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