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Updated: Jul 2, 2025

A Robust Discovery Platform for the Identification of Novel Mediators of Melanoma Metastasis
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Biomarker RIPK3 Is Silenced by Hypermethylation in Melanoma and Epigenetic Editing Reestablishes Its Tumor Suppressor

Sarah Arroyo Villora1, Paula Castellanos Silva1, Tamara Zenz1

  • 1Institute for Genetics, Justus-Liebig-University Giessen, 35390 Giessen, Germany.

Genes
|February 24, 2024
PubMed

Insights

Receptor Interacting Serine/Threonine Protein Kinase 3 (RIPK3) acts as a tumor suppressor in melanoma. Epigenetic inactivation via hypermethylation correlates with tumor progression, but CRISPR-based demethylation restores RIPK3 expression and reduces melanoma cell fitness.

Area of Science:

  • Cancer Biology
  • Epigenetics
  • Melanoma Research

Background:

  • Cancer remains a leading global cause of death, with epigenetic modifications like DNA hypermethylation contributing to tumorigenesis by inactivating tumor suppressor genes.
  • Hypermethylation of CpG islands near gene promoters significantly impacts gene expression, playing a crucial role in cancer development.

Purpose of the Study:

  • To investigate the epigenetic regulation and inactivation of the potential tumor suppressor gene Receptor Interacting Serine/Threonine Protein Kinase 3 (RIPK3) in melanoma.
  • To explore the functional consequences of RIPK3 epigenetic silencing and its potential as a biomarker.

Main Methods:

  • Bisulfite pyrosequencing was used to detect and quantify RIPK3 CpG island hypermethylation in melanoma samples.
  • A CRISPR/dCas9-based epigenetic editing system (CRISPRa activation) was employed to modulate RIPK3 DNA methylation and expression.
  • Phenotypic characterization of melanoma cells was performed using fluorescence microscopy, flow cytometry, and wound healing assays.

Main Results:

  • A significant correlation was observed between RIPK3 CpG island hypermethylation and reduced RIPK3 expression in melanomas.
  • RIPK3 methylation rates increased with advancing tumor stage, suggesting its involvement in melanoma progression.
  • Epigenetic editing-induced RIPK3 re-expression and demethylation led to reduced melanoma cell fitness and demonstrated its tumor suppressive function.

Conclusions:

  • RIPK3 functions as an epigenetically regulated tumor suppressor in melanoma.
  • RIPK3 epigenetic status can serve as a potential biomarker for melanoma classification and progression.
  • Targeting RIPK3 epigenetic inactivation offers a potential therapeutic strategy for melanoma.
Keywords:
CRISPR-Cas9DNA (hyper)methylationRIPK3biomarkercancerdeactivated dCas9epigenetic editingskin cutaneous melanomatumor suppressor gene

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