# Deciphering the Dynamic Notch Signaling Code

> **NIH NIH R01** · CALIFORNIA INSTITUTE OF TECHNOLOGY · 2020 · $411,353

## Abstract

Abstract
The Notch signaling pathway directs cell fate decisions in diverse tissue contexts, plays key roles in disease,
and represents a major drug target. The pathway uses multiple ligands and receptors that interact with one
another in a promiscuous fashion, as well as Fringe glycosyltransferases that modulate those interactions.
Recent work from our lab suggests that these interactions comprise a communication ‘code’ in which different
ligands activate Notch receptors with distinct dynamics. These dynamics are, in turn, decoded to selectively
activate distinct transcriptional target programs. Current understanding of the code is limited to just two ligands
and one receptor. Determining how dynamic encoding occurs across a broader repertoire of
ligand-receptor-Fringe combinations will enable better understanding, prediction, and control of signaling
interactions between different cell types in diverse developmental and disease contexts. Here, we will combine
cell line engineering, quantitative single-cell time-lapse imaging, direct control of Notch dynamics using mutant
receptors and pharmacological perturbations, and analysis of Notch dynamics in neural stem cells and chick
embryos to decipher this code and its functional roles. In ​Aim 1​, we will focus on encoding, by mapping
dynamic signaling modes across a full matrix of Notch receptor, ligand, and Fringe protein combinations. We
will further extend this approach to analyze co-expression of multiple Notch receptors in the same cell, a
pattern that occurs frequently in natural cell types. In ​Aim 2​, we will focus on decoding, by computationally and
experimentally investigating how cis-regulatory and trans-regulatory mechanisms together enable different
signaling dynamics to selectively activate distinct target gene expression programs. Finally, in ​Aim 3 we will
analyze the function of the Notch dynamic code in neurogenesis, using mouse neural stem cells and chick
embryonic spinal cord development as model systems. In neural stem cells, using a combination of
co-cultures, time-lapse microscopy, and end-point multiplexed single molecule RNA-FISH, we will map
ligand-receptor combinations to Notch activity dynamics, and relate those dynamics in turn to cell fate
decisions. In chick embryos, a Notch-specific fluorescent reporter, together with a new tissue slice preparation
that enables imaging of individual living cells during spinal cord development, will enable us to link Notch
dynamics to cell fate determination. Together, these results will reveal the structure and function of the
dynamic code underlying Notch signaling, and show how it operates in key developmental contexts. More
generally, they should help establish a paradigm for understanding signal encoding and decoding behaviors in
cellular communication systems.

## Key facts

- **NIH application ID:** 9972015
- **Project number:** 2R01HD075335-06A1
- **Recipient organization:** CALIFORNIA INSTITUTE OF TECHNOLOGY
- **Principal Investigator:** MICHAEL B ELOWITZ
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $411,353
- **Award type:** 2
- **Project period:** 2012-09-26 → 2025-02-28

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9972015

## Citation

> US National Institutes of Health, RePORTER application 9972015, Deciphering the Dynamic Notch Signaling Code (2R01HD075335-06A1). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/9972015. Licensed CC0.

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