# Revealing novel mechanisms of gout flares by transcriptional phenotyping synovial and peripheral blood mononuclear cells.

> **NIH NIH R01** · UNIVERSITY OF ALABAMA AT BIRMINGHAM · 2024 · $317,476

## Abstract

Gout is the most common inflammatory arthritis, but beyond the well described role of the NLRP3
inflammasome, the molecular genetic causes of gout flares is least well known among the arthropathies.
Published and preliminary results from our group have implicated genes associated with inflammation in gout
which do (e.g., TMEM176B) or may not (e.g, FADS2) relate directly to inflammasome activation. A powerful
approach for understanding the mechanistic basis of gout flares is through transcriptomic profiling of
mononuclear cells from the target organ of inflammation, the synovial fluid, but these studies have not been
rigorously carried out. We have designed a first of its kind, time-sensitive ancillary study, through the parent
clinical trial, Treat-to-Target vs Treat-to-Avoid-Symptoms (TRUST), which will allow novel comparisons of
candidate gene expression in mononuclear cells of synovial fluid to peripheral blood cells prior to and during
active gout flares. These studies will require consenting TRUST participants for arthrocentesis during active
flares. This additional, fresh biospecimen collection, facilitated by the TRUST trial, is highly time-sensitive and
crucial for thorough investigation of molecular mechanisms of gout flares. We will use these samples to carry
out Aim 1 - differential gene expression analysis of myeloid subsets before and during a flare in peripheral
blood and synovial fluid samples during active flare. Flow cytometry on samples from 20 individuals will be
used to isolate non-classical and classical monocytes followed by bulk RNA sequencing. Single cell RNAseq
will be done on peripheral blood mononuclear cell (PBMC) samples from five of the 20 individuals to infer
specific cell-types. In Aim 2, using our existing ex vivo cell culture paradigm we will stimulate PBMCs with LPS
and C18.0 fatty acid (FA) in the presence/absence of MSU crystals, followed by scRNAseq 24 hours post
stimulation. Cells will be isolated from fresh samples of blood collected from people with gout (intercritical)
enrolled at UAB. scRNAseq transcriptomes will be compared between unstimulated, C18.0 FA and LPS
stimulated cells. Readouts will allow mechanistic insight into gout flares that are TLR-agonist mediated and
related to potential gout flare triggers. Finally, we will knockdown candidate genes, by attenuating their
expression and then measure NLRP3 inflammasome activation. Altered expression of secreted IL-1β would
confirm their mechanistic role in gout flares. The proposed research is highly innovative by sampling, with
never before collections of matched samples of peripheral blood and synovial fluid, by analyses, novel
transcriptomic analyses of myeloid cells in samples and in an ex vivo system, and by design, leveraging
resources of the TRUST parent clinical design for this time-sensitive ancillary study. Our work will provide new
knowledge of genetically-based mechanisms of gout flares and aid in identification of new therapeutic targets.

## Key facts

- **NIH application ID:** 10995564
- **Project number:** 1R01AR084827-01
- **Recipient organization:** UNIVERSITY OF ALABAMA AT BIRMINGHAM
- **Principal Investigator:** TONY R MERRIMAN
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $317,476
- **Award type:** 1
- **Project period:** 2024-09-18 → 2028-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10995564

## Citation

> US National Institutes of Health, RePORTER application 10995564, Revealing novel mechanisms of gout flares by transcriptional phenotyping synovial and peripheral blood mononuclear cells. (1R01AR084827-01). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10995564. Licensed CC0.

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