# Single Cell Analysis of MAPK Signaling Dynamics during Tissue Homeostasis

> **NIH NIH R35** · JOHNS HOPKINS UNIVERSITY · 2020 · $480,369

## Abstract

Project Summary
The long term goal of our research is to understand how MAPK signaling dynamics controls and coordinates
cell fate during tissue homeostasis and development. Mitogen Activated Protein Kinases (MAPKs) are clinically
relevant signaling molecules that orchestrate cellular responses to a diverse array of stimuli. There are three
major MAPK signaling cascades (ERK, p38 and JNK) that control opposing cellular decisions such as
survival/apoptosis or proliferation/senescence. Even though these opposed functional roles have been well-
characterized, a wide variety of stimuli (i.e. cytokines, cellular stresses or growth factors) have been shown to
activate all branches of this highly interconnected network. In addition, whether cells finally apoptose, senesce
or enter cell cycle is a highly heterogeneous outcome, even in isogenic cells experiencing the same
environment. Our current understanding of how the MAPK network controls cell fate is incomplete because: (i)
a lack of integrated methods to quantify the dynamics of the network as a whole and (ii) the use of cell
population assays that average unsynchronized single cell behaviors.
To address this need, my laboratory has pioneered a new generation of biosensors that allow simultaneous
quantification of multiple kinase activities in thousands of live single cells. These biosensors convert
phosphorylation into a nucleocytoplasmic shuttling event that can be easily measured by fluorescent
microscopy. Our unique methodology features the high temporal resolution, sensor multiplexing capabilities,
and single cell resolution essential for studying signaling network dynamics in single cells of a multicellular
system.
Our central hypothesis is that the signaling equilibrium between MAPKs is critical to regulate single cell
outcomes (i.e. proliferation, quiescence, senescence, apoptosis). However, the crosstalk dynamics between
individual MAP kinases has not been systematically studied. This is, in part, because MAPK studies use a wide
variety of experimental conditions, cell types and genetically altered cells. In this project we will dissect MAPK
signaling dynamics at the molecular, cellular and multicellular levels: In Project 1 we will systematically
interrogate the crosstalk dynamics between MAP kinases and the phenotypic consequences of this crosstalk.
In Project 2 we will use cell culture and primary organoid models to understand the role of MAPK signaling in
maintaining tissue homeostasis during early oncogenesis. In Project 3 we will study how MAPK signaling
dynamics robustly specifies the mammalian embryo during preimplantation development.

## Key facts

- **NIH application ID:** 9990815
- **Project number:** 5R35GM133499-02
- **Recipient organization:** JOHNS HOPKINS UNIVERSITY
- **Principal Investigator:** Sergi Regot
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $480,369
- **Award type:** 5
- **Project period:** 2019-08-07 → 2024-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9990815

## Citation

> US National Institutes of Health, RePORTER application 9990815, Single Cell Analysis of MAPK Signaling Dynamics during Tissue Homeostasis (5R35GM133499-02). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/9990815. Licensed CC0.

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