# Spatiotemporal tools to interrogate O-GlcNAc functions in cellular signaling

> **NIH NIH R35** · WAYNE STATE UNIVERSITY · 2022 · $365,743

## Abstract

Abstract
The overall goal of the Fehl laboratory is to develop chemical biology strategies to determine the functional
impact of protein modifications during signaling processes. Specifically, cellular metabolism and stress each lead
to diverse protein modifications with O-linked N-acetylglucosamine sugar (O-GlcNAc) but no tools are currently
able to capture highly dynamic and transient O-GlcNAc events with defined time and spatial resolution. Lack of
“time and space” rigor hinders the scientific community from connecting metabolism with disease physiology,
including significantly elevated cancer risk in diabetic patients observed for malignancies like breast cancer.
In this MIRA application, we pose our strategies to address this critical gap through the development of real-time
and space molecular tools that bridge cell metabolism and cancer processes using O-GlcNAc as the keystone.
Excellent NIH-funded research has discovered over 2000 O-GlcNAc on proteins in human cells, but current tools
rely on disrupted physiology, leading to artifacts, or miss key GlcNAc-driven signaling events that occur before
global metabolic rebalancing occurs in less than an hour. We hypothesize that the key drivers of hyperglycemic
metabolism and pathology lie within the first few minutes of nutrient and signaling stimulation, which to date is
not possible to observe in living cells. Our published work in photochemistry and systems glycobiology support
our unique strategies to trigger O-GlcNAc processes in minutes, before O-GlcNAc rebalancing occurs. Our
photocaged sugar tool is able to trigger the oncogenic transcription factor NFkB movement between cytosol or
endoplasmic reticulum into the nucleus, simulating physiological events that potentially link aberrant insulin and
glucose release in diabetes with breast cancer risk. Our real time system can be used to track O-GlcNAc events
during insulin signaling for the first time during the rapid, 15-minute pulses of diseased insulin physiology.
Another tool for targeted intracellular O-GlcNAc-targeted proximity labeling is able to track O-GlcNAcylated
proteins in subcellular space, which no reported tool has the capability to specifically label in live cells. We
propose in the next 5 years to develop our “time and space” molecular tools and apply them for unique
mechanistic studies in disease biology through NFkB targeting. We actively collaborate with metabolic disease
and cancer specialists to ensure disease relevance, as well as with industrial scientists for technology
development to expand industrial awareness of O-GlcNAc biology in metabolism-driven disease pathways.
The research outputs of this proposal include molecular probes, spatiotemporal strategies, and targets to
connect cellular metabolism with signaling. Our enabling chemical strategies have the potential for broad impact
in the scientific community by establishing temporal and spatial methods to study protein modifications. Our
platforms can be extende...

## Key facts

- **NIH application ID:** 10414136
- **Project number:** 5R35GM142637-02
- **Recipient organization:** WAYNE STATE UNIVERSITY
- **Principal Investigator:** Charlie Fehl
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $365,743
- **Award type:** 5
- **Project period:** 2021-07-01 → 2026-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10414136

## Citation

> US National Institutes of Health, RePORTER application 10414136, Spatiotemporal tools to interrogate O-GlcNAc functions in cellular signaling (5R35GM142637-02). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10414136. Licensed CC0.

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