# Transcriptional Control During Erythropoiesis

> **NIH NIH R01** · INSTITUTE FOR SYSTEMS BIOLOGY · 2021 · $668,311

## Abstract

Erythropoiesis is a dynamic process governed by quantitative changes in the relative levels of transcription fac-
tors (TFs). Due to the current paucity of quantitative data on the proteins that constitute the transcriptional regu-
latory network (TRN), most models of erythropoiesis are based primarily on mRNA measurements and do not
typically consider changes in the protein levels of specific TFs. This significantly limits the understanding of
erythropoiesis and other transcriptionally regulated processes such as ß-globin expression, ultimately impinging
on the capacity to correct hemoglobin disorders. The long-term goal is to decipher the TRN that controls eryth-
ropoiesis in health and disease. The objective of this proposal is to significantly expand our TRN model for cell
fate decision during erythropoiesis by integrating dynamic bulk and single cell TF protein abundance measure-
ments with other transcription-relevant -omics data. The central hypothesis is that the relative protein levels of
TFs are critical parameters in the establishment of proper gene expression programs during the continuum of
differentiation, and that erythropoiesis is driven by graded changes in the relative amounts of specific combina-
tions of TFs. The rationale is that integration of the dynamic and quantitative nature of the TF proteome into an
expanded TRN of erythropoiesis will result in a model with improved predictive power which will serve as a
benchmark for healthy erythropoiesis against which to compare erythroid-related disease states, and will facili-
tate the identification of pharmacological agents to restore normal erythropoiesis. Three specific aims have been
designed: 1) Absolute quantification of the TF proteome during erythropoiesis; 2) Determine how gradual
changes in the abundance of multiple TFs in single cells initiate and progressively reinforce cell fate decisions
along the erythroid trajectory; and 3) Computational analysis, modeling and validation of the erythropoiesis TRN.
For the first aim, a high throughput quantitative mass spectrometry (MS) approach will be used to measure
absolute levels of the TF proteome during ex vivo erythropoiesis of HSPCs derived from healthy donors. For the
second aim, complementary CyTOF and targeted-MS proteomic approaches will be combined to estimate TF
protein abundances in single cells, with other single cell –omics technologies to measure changes in gene ex-
pression and TF genomic binding during ex vivo erythropoiesis. Under the third aim, TRN models of erythropoi-
esis will be built utilizing measurements of TF protein abundances, and other transcription-relevant –omics data.
Functional validation will be performed for TFs that have been implicated in transcriptional control during eryth-
ropoiesis based on our recent results. The approach is innovative because it uses a novel combination of single
cell and bulk proteomics methodologies to quantify large numbers of TFs during erythropoiesis in primary h...

## Key facts

- **NIH application ID:** 10200020
- **Project number:** 5R01DK098449-07
- **Recipient organization:** INSTITUTE FOR SYSTEMS BIOLOGY
- **Principal Investigator:** Marjorie Carole Brand
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $668,311
- **Award type:** 5
- **Project period:** 2013-09-16 → 2025-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10200020

## Citation

> US National Institutes of Health, RePORTER application 10200020, Transcriptional Control During Erythropoiesis (5R01DK098449-07). Retrieved via AI Analytics 2026-05-21 from https://api.ai-analytics.org/grant/nih/10200020. Licensed CC0.

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