# Defining the function of SETD8 in colorectal cancer

> **NIH NIH F31** · UNIVERSITY OF PENNSYLVANIA · 2022 · $34,179

## Abstract

Project Summary
 Colorectal cancer (CRC) is predicted to be the second leading cause of cancer-related deaths in 2018.
Greater than 90% of CRC tumors contain a mutation in the canonical Wnt-signaling pathway, which deregulates
β-CATENIN-TCF transcriptional activity and drives oncogenic gene expression. Indeed, the canonical Wnt-target
LGR5 is proposed mark CRC stem cells, which are required for primary and metastatic tumor growth. In addition
to genetic mutations, CRC tumors bear alterations in epigenetic features, including global DNA hypomethylation,
CpG island hypermethylation and a variety of changes in histone post-translational modifications. Although these
chromatin changes are thought to be important for CRC tumorigenesis, the oncogenic chromatin factors in CRC
remain largely undefined, impeding targeting of epigenetic processes in the clinic.
 To address this gap in knowledge, I conducted an in vitro CRISPR-Cas9 dropout screen using a
chromatin-focused pooled lentiviral guide RNA (gRNA) library in a primary murine CRC model. gRNAs targeting
Kmt5a (encoding SETD8), the sole histone 4 lysine 20 (H4K20) monomethyltransferase, dropped out
significantly during this screen, indicating the importance of this gene in CRC proliferation. SETD8 and
H4K20me1 play important roles in regulating transcription, DNA damage repair and cell cycle progression.
Particularly relevant for CRC, SETD8 methyltransferase activity is required for β-CATENIN-TCF transcriptional
function in human cell lines, zebrafish and Drosophila. Indeed, upon Wnt stimulation, SETD8 directly interacts
with β-CATENIN-TCF complexes and deposits H4K20 monomethyl (H4K20me1) modifications at canonical Wnt
target genes. Based on these findings, I hypothesize that SETD8 promotes CRC tumorigenesis in part by
cooperating with β-CATENIN-TCF to potentiate the canonical Wnt-signaling pathway. To this end, in Aim 1 I will
investigate the importance of SETD8 in CRC tumorigenesis in vivo by manipulating SETD8 expression in murine
CRC organoids and orthotopically implanting these lines into a syngeneic host. Moreover, I will define an in vitro
therapeutic index for the SETD8 inhibitor UNC-0379 in pairs of primary normal and CRC organoids derived from
human patients. In Aim 2 I will elucidate the mechanistic role of SETD8 in β-CATENIN-TCF transcriptional
activity by manipulating SETD8 levels and measuring canonical Wnt-target activation by transcriptome profiling
and functional reporter assays, as well as by quantifying the abundance and activity of Lgr5+ CRC stem cells in
vivo. I will also define the protein-protein interactions between β-CATENIN, the TCF factor LEF1 and SETD8 as
well as the enrichment of these proteins and H4K20me1 at canonical Wnt-target loci.
 This study will determine, for the first time, the role of SETD8 in CRC tumorigenesis. The proposed
experiments will potentially identify a promising therapeutic avenue to combat this deadly disease.

## Key facts

- **NIH application ID:** 10359781
- **Project number:** 5F31CA250267-03
- **Recipient organization:** UNIVERSITY OF PENNSYLVANIA
- **Principal Investigator:** ZVI Cramer
- **Activity code:** F31 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $34,179
- **Award type:** 5
- **Project period:** 2020-03-01 → 2023-02-28

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10359781

## Citation

> US National Institutes of Health, RePORTER application 10359781, Defining the function of SETD8 in colorectal cancer (5F31CA250267-03). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10359781. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*