# Motoki Project

> **NIH NIH P20** · UNIVERSITY OF NORTH DAKOTA · 2020 · $204,634

## Abstract

PROJECT SUMMARY. PROJECT 5, M.TAKAKU.
Pioneer transcription factors are capable of binding to closed chromatin and inducing its opening to drive cell
reprogramming. The mechanisms of pioneer factors’ actions remain largely unknown. Using a model of
mesenchymal-to-epithelial transition (MET) in human breast cancer cells, we have discovered that a pioneer
factor GATA3 can open chromatin only at a subset of its binding sites. Our central hypothesis is that GATA3
requires appropriate chromatin context to achieve gene activation that are essential for GATA3 mediated cell
reprogramming. The overall objectives in this grant are to identify chromatin structures and GATA3 co-factors
needed for GATA3-driven chromatin opening and cell reprogramming. The central hypothesis will be tested by
pursuing two specific aims: (1) Define the specific chromatin structures required for GATA3 driven MET; and
(2) Identify the roles of GATA3 co-factors for epithelial cancer cell reprogramming. We will use in vitro and in
vivo breast cancer cell systems to address these specific aims. Under the first aim, the doxycycline inducible
GATA3 expression system in MDA-MB-231 cells that we have established will be used to identify the roles of
nucleosome positioning and linker histones (H1.3 and H1.5) during GATA3-mediated MET. Genomic
techniques including ChIP-seq, ATAC-seq, RNA-seq and MNase-seq together with CRISPR-Cas9 genome
editing and shRNA knockdown methods will be utilized. For the second aim, the same inducible GATA3
expression system and a GATA3 mutant T47D cell line (established by our group) will be used to identify the
roles of chromatin remodeling factors BRG1 and CHD4 in the GATA3-mediated MET and the GATA3 mutant
induced epithelial-to-mesenchymal transition (EMT). It is important to understand the roles of GATA3 co-
factors in the GATA3 mutant cell context, because GATA3 was recently identified as one of the most frequently
mutated genes in breast cancer. Under both aims, mouse xenograft tumor model will be used to identify in vivo
roles of chromatin structures and chromatin remodeling factors in tumor growth and metastasis. The proposed
research is innovative in two ways: first, it features novel hypotheses that will advance the fields of
transcription, chromatin and cancer biology, and secondly, there are multiple innovative technologies that are
proposed, most of which the project leader has developed. The proposed research is significant because it is
expected to provide a new framework for understanding the functions of transcription factors, chromatin
remodeling factors, and chromatin structures in cancer cell reprogramming. Ultimately, such knowledge has
the potential to offer development of new therapeutic strategies of breast cancer treatment.

## Key facts

- **NIH application ID:** 9976557
- **Project number:** 5P20GM104360-07
- **Recipient organization:** UNIVERSITY OF NORTH DAKOTA
- **Principal Investigator:** Motoki Takaku
- **Activity code:** P20 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $204,634
- **Award type:** 5
- **Project period:** — → —

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9976557

## Citation

> US National Institutes of Health, RePORTER application 9976557, Motoki Project (5P20GM104360-07). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/9976557. Licensed CC0.

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