# Molecular regulation of hepatic cell differentiation and maturation

> **NIH NIH R01** · CINCINNATI CHILDRENS HOSP MED CTR · 2020 · $402,805

## Abstract

PROJECT SUMMARY
Our application builds on the published knowledge that during embryonic development until after birth,
specified hepatocytes undergo a process of differentiation where they adopt the physiological functions and
morphology associated with the adult liver. Our preliminary studies deleting Zinc finger HIT-type containing 1
(Znhit1) at mid-gestation in embryonic hepatoblasts demonstrates a specific and essential role in the postnatal
liver for survival, normal cellular architecture, and molecular gene signatures without changing the expression
of the six key hepatic cell master regulators. Znhit1's proposed role as a core subunit of the Snf2-Related
CREB-binding protein Activator Protein (SRCAP)-chromatin remodeling complex may initiate the necessary
changes in chromatin architecture through insertion of the alternative histone H2A.Z to influence gene
expression. The goal of this application is to determine whether Znhit1 allows or disrupts access of
transcription factors to different gene targets and/or enhancers, and thereby provides a switch towards hepatic
cell differentiation. We hypothesize that Znhit1, part of the SRCAP complex, regulates the hepatocyte
differentiation-dependent transcriptional program. In Aim 1 we will determine whether Znhit1's impact on liver
function is due to 1) an autonomous hepatoblast and/or hepatocyte defect and 2) master regulator access to
gene targets and/or enhancers. For this we will use in vivo mouse models to perform temporal hepatoblast and
hepatocyte deletion of Znhit1, and to determine if the hepatoblast and/or hepatocyte transcriptional program
and genomic binding sites of master regulators such as Hnf4a and Foxa2 are impacted. In Aim 2 we will define
what regulates the developmental switch for hepatocyte adult master regulator function. To determine whether
Znhit1 is part of the switch to enable repression of the fetal and/or activation of the adult hepatocyte program,
we will use AAV/DJ8-Ttr-Znhit1-GPF to force expression of Znhit1 at specific time points in mouse models and
in culture iPSC-generated hepatocyte-like cells. In Aim 3, we will determine whether H2A.Z is incorporated into
specific sites in a differentiation-dependent manner regulated by Znhit1. To answer this question, we will use a
Znhit1-3xFlag-P2A-Zsgreen knock-in mouse to perform Znhit1 ChIP-Seq and compare to Znhit1 dependent
H2A.Z bound sites. Our research will generate new insight into the molecular regulation of hepatic cell identity
and differentiation.

## Key facts

- **NIH application ID:** 10022327
- **Project number:** 5R01DK120765-02
- **Recipient organization:** CINCINNATI CHILDRENS HOSP MED CTR
- **Principal Investigator:** Stacey S Huppert
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $402,805
- **Award type:** 5
- **Project period:** 2019-09-23 → 2023-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10022327

## Citation

> US National Institutes of Health, RePORTER application 10022327, Molecular regulation of hepatic cell differentiation and maturation (5R01DK120765-02). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10022327. Licensed CC0.

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