# Systems analysis of cell-to-cell variability and signaling-transcription factor motifs regulating macrophage responses to conflicting environmental cues

> **NIH NIH R01** · YALE UNIVERSITY · 2022 · $334,944

## Abstract

PROJECT SUMMARY
Cell-to-cell variability provides a strategy by which cells can explore diverse states; little is known,
however, about the regulatory motifs that stabilize particular states into cell subsets that enable
diverse functions within a population. Macrophages, cells of the innate immune system, provide
an ideal experimental system to study the regulation of cell subsets, because they transiently
adopt polarization states (i.e., tailored sets of molecules) to meet changing functional demands in
tissues. However, the polarization state adopted to perform one task is often suboptimal for–or
even in opposition to–performing other tasks, raising the question of how macrophages respond
to conflicting microenvironmental cues. Our recent work suggests that cell-to-cell variation in
mutually inhibitory transcription factor (TF) networks increases macrophage functional diversity
and enables them to meet functional demands encoded by conflicting cues. The overall objective
of this proposal is to define intracellular and extracellular signaling–TF networks that regulate
heterogeneous pro-inflammatory (e.g., in response to LPS+IFN-γ) vs. anti-inflammatory (e.g., in
response to IL-4) macrophage subsets and ultimately coordinate a coherent functional response.
Our central hypothesis is that mutually inhibitory networks establish macrophage pro- vs. anti-
inflammatory subsets that are coordinated via context-dependent mechanisms. To test this
hypothesis, we will integrate single-cell measurements with mathematical modeling to determine
if mutual inhibition and collective sensing are important network motifs for harnessing cell-to-cell
variability and organizing appropriate polarization states to meet functional demands in tissues.
Achieving the proposed aims would provide an alternative way to describe the macrophage
polarization spectrum in innate immunity, by conceptualizing it as a set of functional cell subsets
that emerge from multiple competing network motifs. The proposed research is innovative
because we will measure single-cell responses after co-stimulation with conflicting environmental
cues–specifically, cues that have known mechanisms of regulatory cross talk and cell-cell
communication–in order discover new regulatory mechanisms and establish new frameworks
describing macrophage functional diversity. The proposed research is significant because it is
expected to have broad translational potential to identify therapeutic targets that more specifically
alter macrophage functional states in vivo to treat conditions characterized by dysregulation of
macrophage functions.

## Key facts

- **NIH application ID:** 10445622
- **Project number:** 2R01GM123011-05
- **Recipient organization:** YALE UNIVERSITY
- **Principal Investigator:** KATHRYN MILLER-JENSEN
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $334,944
- **Award type:** 2
- **Project period:** 2017-03-01 → 2026-01-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10445622

## Citation

> US National Institutes of Health, RePORTER application 10445622, Systems analysis of cell-to-cell variability and signaling-transcription factor motifs regulating macrophage responses to conflicting environmental cues (2R01GM123011-05). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10445622. Licensed CC0.

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