# DNMT3B Control of Melanoma Heterogeneity and Anti-Tumor Immunity

> **NIH NIH F31** · YALE UNIVERSITY · 2020 · $21,863

## Abstract

ABSTRACT
Melanoma is the deadliest form of skin cancer, and accounts for nearly 8,000 deaths each year in the United
States. Recently developed immune checkpoint therapies including α-CTLA4 and α-PD1 are promising
strategies for treating melanoma and have produced impressive clinical results. However, many patients on
these therapies eventually develop resistance, and there remains no cure for melanoma. Aberrant
methylation has long been recognized as a driver of melanoma progression and is associated with poor
survival in patient cohorts. Increased DNMT3B expression is associated with poor survival in patient cohorts,
and DNMT3B promotes melanoma progression via miR-196b and the mTORC2 pathway. Preliminary data
indicates that DNMT3B modulates phenotypically distinct cell subsets in murine and human melanoma.
Furthermore, hypomethylation of the immune checkpoint PD-L1 was recently shown to be associated with
improved survival in melanoma patients, suggesting a role for DNA methylation in the anti-tumor immune
response. Therefore, we hypothesize that DNMT3B regulates intratumoral heterogeneity and anti-tumor
immunity in melanoma. In our first aim, we will test the heterogeneity of melanoma cell lines lacking
DNMT3B. We will then use bisulfite sequencing and expression assays to evaluate methylation changes
between subsets in knockout- and wild-type cells. We will use CRISPR/Cas9-mediated genetic knockouts to
validate putative mechanisms by which DNMT3B regulates heterogeneity in human and murine cell lines. In
our second aim, we will assess DNMT3B control of anti-tumor immunity by evaluating growth of DNMT3B-null
and wild-type melanoma tumors in immune-competent and deficient host animals, and assess associated
tumor microenvironment changes. We will then identify methylation and expression changes in DNMT3B-null
tumors, and use genetic knockouts to validate putative mechanisms. Finally, we will use human datasets and
cell lines to translate these findings into human melanoma. Together, these studies will identify mechanisms
by which DNMT3B promotes melanoma heterogeneity and resistance to immune checkpoint blockade. This
may lead to development of therapies targeting these mechanisms, for improving survival of patients with
advanced melanoma.

## Key facts

- **NIH application ID:** 9990333
- **Project number:** 1F31CA243212-01A1
- **Recipient organization:** YALE UNIVERSITY
- **Principal Investigator:** Meaghan K. Van Den Eynde
- **Activity code:** F31 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $21,863
- **Award type:** 1
- **Project period:** 2020-04-01 → 2020-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9990333

## Citation

> US National Institutes of Health, RePORTER application 9990333, DNMT3B Control of Melanoma Heterogeneity and Anti-Tumor Immunity (1F31CA243212-01A1). Retrieved via AI Analytics 2026-06-12 from https://api.ai-analytics.org/grant/nih/9990333. Licensed CC0.

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