# The role of gene enhancer elements in colon cancer

> **NIH NIH R01** · CASE WESTERN RESERVE UNIVERSITY · 2021 · $75,480

## Abstract

PROJECT(SUMMARY/ABSTRACT!
Colorectal cancer (CRC) is associated with the deaths of over 50,000 adult Americans annually.
In the previous award period, we showed that in addition to genetic mutation, disease
progression is accompanied by epigenetic changes at gene enhancer elements that switch
genes on and off. We term these Variant Enhancer Loci, or VELs. Remarkably, colon tumors
from different individuals show a common pattern of enhancers that are recurrently activated
across patient samples. These recurrently activated enhancers constitute a signature of CRC. In
this competing renewal application, we will investigate three non-mutually exclusive hypotheses
to uncover the mechanism by which these signature VELs form in CRC. Specific Aim 1 tests
the hypothesis that the VELs are a direct consequence of mutations in canonical CRC
oncogenes and tumor suppressors. This hypothesis will be tested through H3K27ac ChIP-seq
analysis of the enhancer epigenome in human intestinal organoids in which each of known CRC
driver genes were sequentially mutated via CRISPR/Cas9 to recapitulate the adenoma-
carcinoma sequence predicted by the Vogelgram. Specific Aim 2 tests the hypothesis that
transcription factors drive formation of the signature VELs. This hypothesis will be tested
through knockdown and overexpression of transcription factors that bind to the signature VELs
in CRC cell lines and intestinal organoid models, followed by analysis of chromatin at CRC
signature VELs. Specific Aim 3 tests the hypothesis that somatic indel mutations in enhancer
elements drive VEL formation. CRISPR-Cas9 genome editing strategies will be used to correct
or introduce candidate enhancer-creating indel mutations in CRC cell lines, followed by
functional analysis of enhancer activity. Lastly, we propose a fourth Aim to assess whether
VELs are required for tumorigenicity of CRC. CRISPR-Cas9-based strategies will be used to
disrupt signature VELs in CRC cell lines, followed by quantification of their growth in mouse
xenografts relative to unedited control cells. We expect to gain fundamental insights to
epigenetic enhancer dysregulation as a root cause of CRC tumorigenesis that could lay the
foundation for targeted therapies for patients.
!

## Key facts

- **NIH application ID:** 10074536
- **Project number:** 5R01CA160356-09
- **Recipient organization:** CASE WESTERN RESERVE UNIVERSITY
- **Principal Investigator:** Peter Christopher Scacheri
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $75,480
- **Award type:** 5
- **Project period:** 2012-04-01 → 2023-12-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10074536

## Citation

> US National Institutes of Health, RePORTER application 10074536, The role of gene enhancer elements in colon cancer (5R01CA160356-09). Retrieved via AI Analytics 2026-05-21 from https://api.ai-analytics.org/grant/nih/10074536. Licensed CC0.

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