# Elucidating the role of H3K27me3 demethylation in the intestinal epithelium

> **NIH NIH F31** · UNIVERSITY OF PENNSYLVANIA · 2022 · $24,049

## Abstract

Project Summary
 The mammalian intestine is an ideal model system to study the molecular events that control cell fate
specification during development and adult homeostasis. During perinatal development of the intestinal
epithelium, proliferative intervillus domains invaginate into the underlying mesenchyme to produce crypts of
Lieberkühn. Concomitant with crypt formation is the emergence of mature Lgr5-expressing intestinal stem cells
(ISCs), which situate at base of these crypts and divide daily during adult homeostasis to self-renew and
produce committed progenitor cells. As progenitor cells migrate up along the crypt walls, they divide further
and undergo progressive differentiation. Upon exiting the crypt and entering villus projections, progenitor cells
complete terminal differentiation and produce mature absorptive and secretory cells. Despite advances in
identifying the transcriptional and molecular mechanisms that facilitate ISC development and differentiation,
the epigenetic mechanisms directing these processes remain poorly characterized. The proposed study will
therefore examine the role of UTX and JMJD3, two histone demethylases, in the intestinal epithelium. UTX and
JMJD3 demethylate trimethylated histone 3 lysine 27 (H3K27me3), a histone modification that is required for
gene silencing. By removing this repressive histone mark, UTX and JMJD3 de-repress lineage defining
transcription factors to promote differentiation during development. Furthermore, UTX and JMJD3 facilitate
differentiation in other regenerative adult tissues and are therefore critical for maintaining adult tissue
homeostasis. Whether UTX and JMJD3 are important epigenetic regulators of differentiation in the intestinal
epithelium remains unknown. Aim 1 will test the hypothesis that UTX- and JMJD3-mediated H3K27me3
demethylation activates a mature stem cell transcriptional program in fetal progenitor cells and is therefore
required for perinatal intestinal development. A mouse model of constitutive, intestine-specific Utx and Jmjd3
ablation will be employed to determine whether UTX and JMJD3 promote perinatal crypt formation and ISC
development. To validate and extend in vivo results, an in vitro organoid formation assay will identify whether
UTX and JMJD3 are required for the maturation of fetal intestinal spheroids into mature budding organoids.
Furthermore, single-cell RNA-Sequencing (scRNA-Seq) and ChIP-Seq will be used to identify transcriptional
and epigenomic changes induced in the perinatal intestine following UTX and JMJD3 ablation. Aim 2 will test
the hypothesis that UTX- and JMJD3-mediated H3K27me3 demethylation is required to activate differentiation
genes and maintain proper differentiation of adult intestinal epithelial cells. Utx and Jmjd3 expression will be
conditionally ablated within the intestinal epithelium of adult mice. These mice will be analyzed by
immunohistochemistry, scRNA-Seq, and ChIP-Seq to determine whether the differentiation program...

## Key facts

- **NIH application ID:** 10396126
- **Project number:** 5F31DK126404-03
- **Recipient organization:** UNIVERSITY OF PENNSYLVANIA
- **Principal Investigator:** Hannah Kolev
- **Activity code:** F31 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $24,049
- **Award type:** 5
- **Project period:** 2020-06-01 → 2023-01-12

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10396126

## Citation

> US National Institutes of Health, RePORTER application 10396126, Elucidating the role of H3K27me3 demethylation in the intestinal epithelium (5F31DK126404-03). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10396126. Licensed CC0.

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