Epigenetic siRNA and Chemical Screens Identify SETD8 Inhibition as a Therapeutic Strategy for p53 Activation in High-Risk Neuroblastoma

Veronica Veschi; Zhihui Liu; Ty C Voss; Laurent Ozbun; Berkley Gryder; Chunhua Yan; Ying Hu; Anqi Ma; Jian Jin; Sharlyn J Mazur; Norris Lam; Barbara K Souza; Giuseppe Giannini; Gordon L Hager; Cheryl H Arrowsmith; Javed Khan; Ettore Appella; Carol J Thiele

doi:10.1016/j.ccell.2016.12.002

Epigenetic siRNA and Chemical Screens Identify SETD8 Inhibition as a Therapeutic Strategy for p53 Activation in High-Risk Neuroblastoma

Cancer Cell. 2017 Jan 9;31(1):50-63. doi: 10.1016/j.ccell.2016.12.002.

Authors

Veronica Veschi¹, Zhihui Liu¹, Ty C Voss², Laurent Ozbun², Berkley Gryder³, Chunhua Yan⁴, Ying Hu⁴, Anqi Ma⁵, Jian Jin⁵, Sharlyn J Mazur⁶, Norris Lam¹, Barbara K Souza¹, Giuseppe Giannini⁷, Gordon L Hager⁸, Cheryl H Arrowsmith⁹, Javed Khan³, Ettore Appella⁶, Carol J Thiele¹⁰

Affiliations

¹ Cell and Molecular Biology Section, Pediatric Oncology Branch, Center for Cancer Research, National Cancer Institute, CRC, 1-3940, 10 Center Drive MSC-1105, Bethesda, MD 20892, USA.
² High-Throughput Imaging Facility, Center for Cancer Research, National Cancer Institute, Bethesda, MD 20892, USA.
³ Oncogenomics Section, Genetics Branch, Center for Cancer Research, National Cancer Institute, Bethesda, MD 20892, USA.
⁴ Center for Biomedical Informatics and Information Technology, Center for Cancer Research, National Cancer Institute, Rockville, MD 20850, USA.
⁵ Department of Structural and Chemical Biology, Oncological Sciences, Pharmacology and Systems Therapeutics, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
⁶ Chemical Immunology Section, Laboratory of Cell Biology, National Cancer Institute, Bethesda, MD 20892, USA.
⁷ Istituto Pasteur-Fondazione Cenci Bolognetti, Department of Molecular Medicine, University La Sapienza, 00161 Rome, Italy.
⁸ Laboratory of Receptor Biology and Gene Expression, Center for Cancer Research, National Institutes of Health, Bethesda, MD 20892, USA.
⁹ Structural Genomics Consortium, University of Toronto, Toronto, ON M5G 1L7, Canada.
¹⁰ Cell and Molecular Biology Section, Pediatric Oncology Branch, Center for Cancer Research, National Cancer Institute, CRC, 1-3940, 10 Center Drive MSC-1105, Bethesda, MD 20892, USA. Electronic address: [email protected].

Abstract

Given the paucity of druggable mutations in high-risk neuroblastoma (NB), we undertook chromatin-focused small interfering RNA and chemical screens to uncover epigenetic regulators critical for the differentiation block in high-risk NB. High-content Opera imaging identified 53 genes whose loss of expression led to a decrease in NB cell proliferation and 16 also induced differentiation. From these, the secondary chemical screen identified SETD8, the H4^K20me1 methyltransferase, as a druggable NB target. Functional studies revealed that SETD8 ablation rescued the pro-apoptotic and cell-cycle arrest functions of p53 by decreasing p53^K382me1, leading to activation of the p53 canonical pathway. In pre-clinical xenograft NB models, genetic or pharmacological (UNC0379) SETD8 inhibition conferred a significant survival advantage, providing evidence for SETD8 as a therapeutic target in NB.

Keywords: SETD8; differentiation; epigenetics; neuroblastoma; p53; siRNA screen.

Published by Elsevier Inc.

MeSH terms

Cell Differentiation
Cell Proliferation
Epigenesis, Genetic*
Histone-Lysine N-Methyltransferase / antagonists & inhibitors*
Histone-Lysine N-Methyltransferase / genetics
Histone-Lysine N-Methyltransferase / physiology
Humans
Neuroblastoma / drug therapy*
Neuroblastoma / genetics
Neuroblastoma / pathology
Quinazolines / pharmacology
RNA, Small Interfering / genetics*
Tumor Suppressor Protein p53 / analysis
Tumor Suppressor Protein p53 / physiology*

Substances

6,7-dimethoxy-2-(pyrrolidin-1-yl)-N-(5-(pyrrolidin-1-yl)pentyl)quinazolin-4-amine
Quinazolines
RNA, Small Interfering
TP53 protein, human
Tumor Suppressor Protein p53
Histone-Lysine N-Methyltransferase
KMT5A protein, human

Abstract

MeSH terms

Substances

Grants and funding