# Cell-based high-throughput chemical genetic screening for GSDMD inhibitors

> **NIH NIH R01** · BOSTON CHILDREN'S HOSPITAL · 2021 · $442,500

## Abstract

Project Summary
Sepsis is a major health threat and the primary cause of morbidity and mortality in the critically ill patients with
infection or trauma. Current treatment for sepsis is mainly supportive, including broad spectrum antibiotics,
fluid resuscitation and vasopressor. Over the last 40 years, numerous attempts have been made to identify
more specific and effective therapeutic strategies and pharmacological agents for treatment of sepsis. At
present, there have been at least 150 clinical trials targeting various components in sepsis induction, including
pathogen-associated molecular patterns (PAMPs), pattern-recognition receptors, and early cytokines or
mediators produced in response to sepsis such as IL-1, TNF, C5a, and activated protein C. None has proven
effective to date. Thus there is a critical need for targeted and effective sepsis therapy to reduce mortality from
this disease. Most recent studies identified Gasdermin D (GSDMD) as a druggable target for the treatment of
sepsis; however no specific inhibitors have been developed for this pathway. The objective of our research is
to identify specific small molecule inhibitors of GSDMD via conducting a high throughput chemical genetic
screening. The hypothesis is that these GSDMD inhibitors would alleviate both the early septic shock caused
by systemic hyperinflammation and the following lethal secondary infections caused by immunosuppression,
and thus should be potential pharmacological agents for preventing/treating sepsis. We have established a
novel cell-based assay to measure and visualize GSDMD-induced lytic cell death. The preliminary data
demonstrated the selectivity, reproducibility, and feasibility of our cell-based assay for high throughput
screening. In this proposed research, we will first use this cell-based system to conduct a primary high
throughput small molecule screening to identify a group of compounds that inhibit GSDMD-mediated lytic cell
death (Aim I). Next, several secondary screening assays will be performed to confirm the effect of each
positive hit compound identified from the primary screening and to identify the most specific and potent ones
for future characterization (Aim II). In addition, we will directly examine the effect of positive compounds on
macrophage and neutrophil function. The final lead compounds should be those that attenuate pyroptosis-
mediated pro-inflammatory cytokine release from macrophages and delay neutrophil death, but do not affect
GSDMD-independent functions such as phagocytosis and chemotaxis (Aim III). Finally, toward understanding
the mechanism of action of each validated positive hit, we will examine whether the lead compounds can affect
GSDMD-NT oligomerization and plasma membrane localization (Aim IV). GSDMD inhibitors identified in this
study can be directly utilized as starting chemical compounds for novel sepsis drug development. Additionally,
discovery of these inhibitors will greatly facilitate our research on the fun...

## Key facts

- **NIH application ID:** 10072027
- **Project number:** 5R01AI141386-03
- **Recipient organization:** BOSTON CHILDREN'S HOSPITAL
- **Principal Investigator:** Hongbo R Luo
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $442,500
- **Award type:** 5
- **Project period:** 2019-01-02 → 2022-12-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10072027

## Citation

> US National Institutes of Health, RePORTER application 10072027, Cell-based high-throughput chemical genetic screening for GSDMD inhibitors (5R01AI141386-03). Retrieved via AI Analytics 2026-05-21 from https://api.ai-analytics.org/grant/nih/10072027. Licensed CC0.

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