# Chromatin Dynamics During Epithelial Commitment

> **NIH NIH R01** · STANFORD UNIVERSITY · 2024 · $598,859

## Abstract

Project Summary/Abstract
 To treat wounds that won’t heal for patients with Recessive Dystrophic Epidermolysis Bullosa,
we have developed induced pluripotent cell (iPS)-cell derived cGMP compatible, scaleable induced skin
composite (iSC) differentiation system that requires multi-lineage interaction to generate graftable
surface ectoderm (SE), mesoderm/dermis and melanocytes. Work from ARO73170, now in its 5th year,
endeavors to fill gaps in knowledge about the chromatin dynamics and interactions between the
lineages to enhance manufacturing efficacy and reduce line-to-line variability. ARO73170 identified
transcription factors (TFs) TFAP2 and GRHL2 that pattern early SE differentiation. In addition GATA3
and the product of the Xia-Gibbs Syndrome locus Gibbin induce Gibbin-dependent mesoderm (GDM)
that matures the SE stratification program through cross-regulatory signaling. For SE differentiation,
GRHL2 activity is required for TFAP2A chromatin binding and in turn, TFAP2A restricts inappropriate
GRHL2 binding to ectodermal disease-associated loci, but the mechanism of interaction, and how
network signaling modifies its output, remains a gap in knowledge. For GDM differentiation, we found
that wild type GDM can rescue mutant ectoderm, reinforcing the need to identity the cross-regulatory
stratification factors that enhance the graftability of manufactured tissue. While GATA3 binding sites are
known, a gap in knowledge exists how Gibbin accumulates on key developmentally regulated promoters
to activate them. Overall our preliminary data supports the intriguing hypothesis that TFAP2-GRHL2
ectoderm signaling, with reciprocal GATA3-Gibbin-GDM induction and signaling, allows proper iPS-
derived graftable tissue manufacturing. We will focus on key gaps in knowledge as we dissect spatial
and temporal TFAP2/GRHL2-mediated surface ectodermal commitment by dissecting the ectoderm-
mesoderm lineage commitment trajectory, refining spatial constraints to optimize surface
ectoderm/mesoderm commitment, interrogating GRHL2-TFAP2 mutual regulatory interactions, and
modulating the GRHL2-TFAP2 target gene network to improve ectoderm commitment; and by
elucidating GDM functions during skin differentiation through elucidating how Gibbin localizes to
promoters of key developmental regulators, dissecting Gibbin-Dependent mesoderm signaling during
mouse ectodermal development, and validating GDM-dependent factors during iPS-derived skin
manufacturing. Successful completion of this proposal will provide deep mechanistic insights into the
chromatin dynamics of multi-lineage tissue differentiation, highlight candidate in-process biomarkers for
clinical manufacturing of skin, and establish a flexible manufacturing platform for novel cell therapy for a
previously untreatable diseases.

## Key facts

- **NIH application ID:** 10975731
- **Project number:** 2R01AR073170-06A1
- **Recipient organization:** STANFORD UNIVERSITY
- **Principal Investigator:** Anthony E Oro
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $598,859
- **Award type:** 2
- **Project period:** 2024-09-01 → 2029-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10975731

## Citation

> US National Institutes of Health, RePORTER application 10975731, Chromatin Dynamics During Epithelial Commitment (2R01AR073170-06A1). Retrieved via AI Analytics 2026-05-26 from https://api.ai-analytics.org/grant/nih/10975731. Licensed CC0.

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