# Theberge 2020 Admin Supplement Equipment

> **NIH NIH R35** · UNIVERSITY OF WASHINGTON · 2020 · $69,942

## 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. 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.

## Key facts

- **NIH application ID:** 10133448
- **Project number:** 3R35GM128648-02S1
- **Recipient organization:** UNIVERSITY OF WASHINGTON
- **Principal Investigator:** Ashleigh Brooks Theberge
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $69,942
- **Award type:** 3
- **Project period:** 2018-08-01 → 2023-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10133448

## Citation

> US National Institutes of Health, RePORTER application 10133448, Theberge 2020 Admin Supplement Equipment (3R35GM128648-02S1). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10133448. Licensed CC0.

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