# Role of an Integrator-EGR axis in the regulation of myeloid enhancers

> **NIH NIH R01** · WISTAR INSTITUTE · 2022 · $446,500

## Abstract

Project Summary
Role of an Integrator-EGR axis in the regulation of myeloid enhancers
Objective: My research aims to identify how enhancer-based regulation determines monocytic cell fate.
Specifically, we seek to understand the function of a newly identified regulatory axis, comprising the Integrator
complex, the EGR-1/EGR-2 transcription factors and their co-factor NAB2. We aim at determining how these
chromatin regulators: a) prime myeloid enhancers for activation; b) remodel nucleosome accessibility and
reshape 3D genome conformation to impose a monocytic-specific transcriptome. We seek to characterize the
first lineage-specific function of the Integrator protein complex, which was previously believed to serve a
general role in transcription.
Proposed research: The process of myelopoiesis is governed by an elaborate gene expression program that
originates in the bone marrow, and is brought to maturity in the peripheral blood and tissues. The
transcriptional process that generates mature myeloid cells from hematopoietic stem cells (HSCs) is initiated
by sequence-specific transcription factors (TFs) that instruct enhancers to elicit gene activation. However, how
these myeloid TFs engage the basal transcriptional machinery and activate lineage-specific enhancers is
poorly understood. We uncovered that the INTS13 subunit of the Integrator complex plays an essential role in
myelopoiesis and we propose that Integrator carries the previously unidentified ability to target myeloid-specific
TFs and modulate cell fate determination via enhancer regulation. We will molecularly dissect the function of
Integrator, and its functional partners EGR-1/2 and NAB2, in monocytic/macrophagic differentiation by
leveraging our expertise in biochemistry and functional genomics through the following aims. 1) We will define
the enhancer network that determines myelopoiesis. We hypothesize that INTS13 modulates Integrator’s
activity at EGR-targeted enhancers during monocytic commitment. Therefore, we will profile INTS13 and
EGR-1/2 recruitment in human cells, and we will determine the status of targeted enhancers by monitoring
their histone methylation and acetylation levels (H3K27Ac and H3K4Me1). Further, we will evaluate the
transcriptional effect of INTS13, EGR-1/2 and NAB2 depletion on myeloid genes by RNA-seq. 2) We will
determine the changes in chromatin architecture mediated by INTS13, EGR-1/2, and NAB2. We hypothesize
that these chromatin regulators prime enhancers for activation and execute cell fate commitment. Our data
suggest that INTS13, EGR-1/2, and NAB2 control the active/poised status of enhancers and oversee genome
topology during differentiation. We will functionally test our hypothesis by analyzing nucleosome remodeling at
enhancers (pioneer activity) and by determining how Integrator and EGR-1/2 modulate chromosomal looping
and 3D genome conformation using Hi-C and 3C assays. Lastly, we will characterize the biochemical
properties of Integrator to id...

## Key facts

- **NIH application ID:** 10310455
- **Project number:** 5R01HL141326-04
- **Recipient organization:** WISTAR INSTITUTE
- **Principal Investigator:** Alessandro Gardini
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $446,500
- **Award type:** 5
- **Project period:** 2019-02-01 → 2023-11-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10310455

## Citation

> US National Institutes of Health, RePORTER application 10310455, Role of an Integrator-EGR axis in the regulation of myeloid enhancers (5R01HL141326-04). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10310455. Licensed CC0.

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