# Hematopoietic Regulation via GATA Switches

> **NIH NIH R01** · UNIVERSITY OF WISCONSIN-MADISON · 2021 · $58,773

## Abstract

Our efforts to forge principles for how GATA factors control development and function of the hematopoietic
system led to the discovery of conserved Gata2 enhancers (+9.5 and -77) essential for hematopoiesis and other
blood-regulatory enhancers. While genomic studies commonly predict enhancers, there were no reports of
enhancers essential for stem cell genesis or progenitor fate decisions. Strikingly, +9.5 mutations resemble
GATA2 coding mutations in causing human GATA2-deficiency syndrome involving immunodeficiency,
myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML). -77 disruption also causes MDS/AML.
This foundation led clinical centers to analyze -77 and +9.5 genetic variation. We discovered: 1) Whereas +9.5
and -77 confer progenitor cell fate, only +9.5 triggers hematopoietic stem cell genesis. 2) GATA2 loss with EVI1
upregulation is leukemogenic. 3) An ensemble of “+9.5-like” enhancers exist, including one regulating a c-Kit
facilitator that promotes erythrocyte regeneration and survival in severe anemia. 4) GATA2-regulated G-protein-
coupled receptor circuits control hematopoiesis. 5) We described gain-of-function (GOF) GATA2 mutants,
suggesting a paradigm-shift. 6) +9.5 single-nucleotide disease mutation dissociates developmental and
regenerative activities, creating a bone marrow failure predisposition. The disease mutations, innovative mouse
models, rescue assay and multiomics will enable the following aims. Aim 1 will determine how a blood
disease-causing enhancer establishes progenitor cell fate. -77-/- progenitors are defective in erythroid and
granulocytic differentiation, yet competent to generate macrophages. Our proteomic and single-cell
transcriptome data revealed that skewed differentiation involves upregulated interferon (IFN) response proteins.
We hypothesize that convergence of GATA2 and IFN mechanisms is critical in progenitor biology. We will
establish the -77-regulated transcriptome/proteome that controls progenitor fate and address key mechanistic
issues. Aim 2 will elucidate genetic determinants for the function of essential GATA2 enhancers. We
generated compound heterozygous (CH) enhancer mutant mice (-77+/-;+9.5+/-) and discovered that both
enhancers must reside on the same allele to regulate progenitor function. We are unaware of any example of a
dual enhancer requirement for progenitor function, the mechanisms may inform human +9.5 and -77
enhanceropathies, we will elucidate the mechanisms. Aim 3 will test models for how GATA2 human disease
mutants exert gain-of-function activity. We innovated a rigorous rescue assay in which restoring GATA2 in -
77-/- progenitors normalizes skewed differentiation and transcription. Mutants assumed to be loss-of-function
(LOF) surprisingly retained activity to rescue granulopoiesis and select target genes. We will test if all disease
mutants exhibit GOF activity, elucidate the mechanisms and determine if activities parse mutants into groups
that may ultimately inform cli...

## Key facts

- **NIH application ID:** 10329057
- **Project number:** 3R01DK068634-16S1
- **Recipient organization:** UNIVERSITY OF WISCONSIN-MADISON
- **Principal Investigator:** Emery H. Bresnick
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $58,773
- **Award type:** 3
- **Project period:** 2005-05-15 → 2024-11-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10329057

## Citation

> US National Institutes of Health, RePORTER application 10329057, Hematopoietic Regulation via GATA Switches (3R01DK068634-16S1). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10329057. Licensed CC0.

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