# Wnt Signaling in intestinal stem cells, homeostasis, and cancer

> **NIH NIH R01** · BOSTON CHILDREN'S HOSPITAL · 2020 · $578,586

## Abstract

Project Summary/Abstract
Signaling by the Wnt family of secreted proteins through transcription coactivator β-catenin (the Wnt/β-catenin
pathway) plays central roles in regulation of intestinal stem cells (ISCs) and homeostasis of the gastrointestinal
(GI) tract. R-spondin (Rspo) proteins are secreted molecules that enhance Wnt/β-catenin signaling through
stabilizing Wnt receptors, and they exhibit potent stimulation effect on self-renewal and proliferation of ISCs.
Anomaly of Wnt/Rspo signaling leads to GI diseases including colorectal cancer (CRC).
Wnt/β-catenin signaling controls ISCs through an ISC-specific gene expression program, which is driven by the
DNA-bound TCF/LEF (T cell factor/Lymphoid enhancer factor) family of transcription factors in complex with β-
catenin. TCF/β-catenin-mediated transcriptional regulation has been a cornerstone for our understanding of the
Wnt/β-catenin pathway including in ISC regulation and CRC pathogenesis.
To better understand Wnt/Rspo signaling and search for additional potential therapeutic target for CRC
treatment, we performed a functional cDNA expression screen and identified a Zinc-finger (Znf) transcription
factor as a potent stimulators of TCF/β-catenin-dependent transcription. Our preliminary data suggest that this
Znf is required for (i) Wnt/Rspo stimulation of ISC expansion in mouse intestinal organoids; (ii) for TCF/β-
catenin-mediated Wnt target genes/stem cell signature genes in human CRC cell lines; and (iii) for proliferation
of CRC cell lines. Our preliminary data further suggest that the Znf binds to TCF, and co-occupies with TCF/β-
catenin on enhancers/promoters of Wnt target genes in chromatin. Our preliminary findings identify a novel
critical component of Wnt/Rspo signaling in ISCs and CRC cells, and reveal an unappreciated complexity in
the mechanism by which TCF/β-catenin-mediated gene activation is achieved.
We propose three specific aims in this application to investigate the Znf in Wnt/Rspo signaling in ISC regulation
and CRC pathogenesis. In Aim 1 we will define the Znf requirement in TCF/β-catenin-driven transcription via
genome-wide RNA-seq and CHIP-seq techniques, thereby addressing whether this factor is required for all or
a subset of TCF/β-catenin target genes; In Aim 2 we will examine the Znf requirement in human intestinal
organoids and primary CRC organoids, attempting to validate its critical role in human GI and cancer biology;
and in Aim 3 we will generate conditional Znf deletion mutant mice, thereby studying its role in intestinal tissue
homeostasis and tumor formation in vivo.

## Key facts

- **NIH application ID:** 9997915
- **Project number:** 5R01DK121945-02
- **Recipient organization:** BOSTON CHILDREN'S HOSPITAL
- **Principal Investigator:** Xi He
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $578,586
- **Award type:** 5
- **Project period:** 2019-08-20 → 2023-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9997915

## Citation

> US National Institutes of Health, RePORTER application 9997915, Wnt Signaling in intestinal stem cells, homeostasis, and cancer (5R01DK121945-02). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/9997915. Licensed CC0.

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