# Decoding Hydrogen Peroxide Signaling at Cellular Membranes

> **NIH NIH R35** · BOSTON CHILDREN'S HOSPITAL · 2022 · $442,500

## Abstract

PROJECT SUMMARY
Eukaryotic cells employ different mechanisms to sense and respond to their environment and maintain tissue
homeostasis. Cells have evolved strategies to co-opt stable reactive oxygen species such as hydrogen
peroxide (H2O2) as non-transcriptional signaling molecules in order to rapidly respond and adapt to
environmental changes. Numerous examples of the influence of H2O2 signaling have emerged, ranging from
abiotic stress in plants to immune responses in humans. H2O2 signaling and subsequent regulation of target
proteins is therefore an important but still underappreciated biological control mechanism. H2O2-regulated cell
signaling is largely dependent on the presence of redox-sensitive thiol switches in protein cysteine residues,
where the reactivity of these switches is highly dependent on the local H2O2 concentration. Our previous
studies in epithelial cells have shown that key determinants of cellular H2O2 concentrations include generation
of H2O2, by cell membrane surface enzymes such as the NADPH oxidases and permeability across cellular
membranes which can be facilitated by Aquaporin (AQP) channels. A number of cellular processes such as
innate immune signaling, vesicular trafficking and migration have been shown to be regulated by H2O2, but
how cellular membranes allow for specific and privileged signaling by H2O2 remains incompletely understood.
We therefore propose studies that aim to establish general rules and emergent concepts related to H2O2
signals at membranes. Our studies will encompass four major areas of inquiry that seek to address i) How
does plasma membrane permeability to H2O2 influence redox signaling and regulation? ii.) How do H2O2
signals and subsequent regulation of proteins alter essential vesicular trafficking pathways in the cell? iii) How
does H2O2 signaling occur at vesicular membranes? iv.) How does spatial control of cellular H2O2 regulate the
directional migration of cells? To address these questions, we will apply and develop high resolution
quantitative fluorescence imaging to follow the spatial and temporal dynamics of H2O2 signals at membranes,
in combination with proteomic approaches to identify target modified cysteines. Further studies will investigate
how oxidative modifications alter target protein structure, function and localization, constructing a mechanistic
understanding of how H2O2 signals are relayed from cellular membranes. Future studies will build on this
framework to uncover strategies to direct and manipulate H2O2 signals for treatment of human disease.
Integrated to these studies will be the development of novel tools and approaches to study H2O2 signals at
membranes that can be broadly applied for research in this field.

## Key facts

- **NIH application ID:** 10458114
- **Project number:** 5R35GM142683-02
- **Recipient organization:** BOSTON CHILDREN'S HOSPITAL
- **Principal Investigator:** Jay Thiagarajah
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $442,500
- **Award type:** 5
- **Project period:** 2021-08-01 → 2026-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10458114

## Citation

> US National Institutes of Health, RePORTER application 10458114, Decoding Hydrogen Peroxide Signaling at Cellular Membranes (5R35GM142683-02). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10458114. Licensed CC0.

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