# Project 2 New Gene Regulatory Proteins Regulating Erythroid Development, Harvey Lodish

> **NIH NIH P01** · BOSTON CHILDREN'S HOSPITAL · 2021 · $455,747

## Abstract

Project 2: Project Summary
Our goal is to understand how a burst- forming unit erythroid (BFU-E) progenitor integrates multiple
environmental (e.g. low oxygen) and hormonal (e.g. cortisol, PPARα agonists) signals and makes the decision
to undergo a self-renewal or differentiative division; in particular we will elucidate the network of transcription
factors and the genes regulated by these factors that control this key developmental decision. In Aim 1 we will
characterize in detail the transcriptomes as BFU-Es initiate and undergo in culture self-renewal divisions in the
presence of dexamethasone together or not with Prolyl Hydroxylase 2 (PH2) inhibitors or PPARα agonists.
Parallel ChIP-Seq analysis of BFU-E cells initiating self-renewal divisions will identify genes potentially directly
regulated by the glucocorticoid receptor (GR), HIF1α, and PPARα. We will use this information in multiple
bioinformatic approaches to identify additional transcription factors, chromatin- modifying enzymes, other
transcriptional regulatory proteins, and RNA-binding proteins that potentially regulate self-renewal. In this
analysis we also anticipate identifying novel cell surface proteins for isolation of early, mid, and late BFU-Es to
higher levels of purity. In Aim 2 we will use cultures of purified early BFU-Es and a combination of dye staining
to count cell divisions and single cell RNA sequencing to determine whether BFU-E self-renewal divisions, as
well as differentiation divisions toward the CFU-E, progress through successive discrete transcriptional states
or form a continuum of states. These studies will define how transcriptional states and numbers of cell divisions
are coordinated during BFU-E self-renewal divisions and also during differentiation toward CFU-Es. In mouse
BFU-Es we showed that PPARα co-occupies many chromatin sites with the glucocorticoid receptor (GR);
these presumed distal enhancer regions are enriched for DNA binding motifs for Smad 2 and 9 other
transcription factors, many of which are known to play important roles in self-renewal of different stem cells. In
Aim 3 we will use gain- and loss- of-function approaches in vitro and in vivo to test whether Smad 2 and ~5
other selected novel gene regulatory proteins identified in Aims 1 and 2, are important for BFU-E self-renewal.
By determining the genes regulated by these factors, as well as by mapping the binding sites of these
transcription factors and RNA binding proteins that are functionally important in the self-renewal process, we
can construct a wiring diagram of the important genes and proteins in the self-renewal network. In addition, our
understanding of how self-renewal occurs in this population of transit amplifying cells will likely have a wide
impact on understanding of the self-renewal process in other cell lineages. Finally, our understanding of the
self renewal of BFU-E cells and the consequent increase in numbers of red cells produced from each BFU-E
progenitor, will li...

## Key facts

- **NIH application ID:** 10146449
- **Project number:** 5P01HL032262-39
- **Recipient organization:** BOSTON CHILDREN'S HOSPITAL
- **Principal Investigator:** Harvey F Lodish
- **Activity code:** P01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $455,747
- **Award type:** 5
- **Project period:** 1997-07-01 → 2022-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10146449

## Citation

> US National Institutes of Health, RePORTER application 10146449, Project 2 New Gene Regulatory Proteins Regulating Erythroid Development, Harvey Lodish (5P01HL032262-39). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10146449. Licensed CC0.

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