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

> **NIH NIH R35** · UNIVERSITY OF WASHINGTON · 2022 · $217,700

## Abstract

Project Abstract
Small molecule and protein signals provide a rich vocabulary for cellular communication, and upstream changes
in RNA expression help drive the molecular dialogue. The downstream consequences of gene expression
changes and signal molecule production are exquisitely sensitive and vary based on microenvironment, disease
state (including temporal considerations), and individual heterogeneity. For example, gene expression changes
vary temporally in response to disease flares or treatments. Dissecting the basic biology of cell signaling and
molecular pathway changes in disease 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 signaling pathways vary across the human population
and through time within an individual? To address these challenges, the parent R35 award has three stated
goals: 1) Develop novel microscale co- and multi-culture platforms to study soluble factor signaling and use these
tools to elucidate paracrine signaling mechanisms. 2) Develop and validate new readouts for inflammation (e.g.,
fibrosis, vasodilation) and apply these methods to identify key effector molecules and signaling pathways in
inflammation. 3) Develop new analytical methods to stabilize, isolate, and study inflammatory signals. Under the
parent R35 award Goal 3, we developed a novel platform that enables at-home blood sampling and RNA
stabilization, homeRNA. The homeRNA kit contains a commercially available at-home blood collection device
and a custom ‘stabilizer tube’ that our lab engineered to contain a stabilizing solution (e.g., RNAlater to stabilize
RNA in blood). Leveraging our lab’s experience with microfluidics and utilizing passive forces (e.g., capillary flow,
Laplace pressure), we engineered the ‘stabilizer tube’ with an integrated fluidic channel that prevents spillage of
RNAlater while the user handles the kit, but enables the transfer of fluid (e.g., RNAlater solution) when the
stabilizer tube is mated with the blood collection tube. Importantly, homeRNA enables longitudinal studies within
an individual to capture temporal changes in gene expression signatures resulting from disease flares or in
response to treatment. homeRNA enables evaluation of mechanistic hypotheses in human populations,
providing a complement to our lab’s in vitro microfluidic platforms, which use cell lines or primary cells. In this
supplement, we will use homeRNA to study the molecular mechanisms underlying inflammation in women of
understudied, underrepresented, and underreported (U3) populations experiencing post-acute sequelae of
SARS-CoV-2 (PASC, also called ‘long COVID’), ultimately enabling better diagnostic and therapeutic
approaches for PASC. Further, we will establish homeRNA as a broadly applicable research tool for reaching
women of U3 populations.

## Key facts

- **NIH application ID:** 10556928
- **Project number:** 3R35GM128648-05S1
- **Recipient organization:** UNIVERSITY OF WASHINGTON
- **Principal Investigator:** Ashleigh Brooks Theberge
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $217,700
- **Award type:** 3
- **Project period:** 2018-08-01 → 2023-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10556928

## Citation

> US National Institutes of Health, RePORTER application 10556928, Deciphering the role of chemical signals in inflammation with open microfluidic functional assays (3R35GM128648-05S1). Retrieved via AI Analytics 2026-05-27 from https://api.ai-analytics.org/grant/nih/10556928. Licensed CC0.

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