Combined inhibition of histone methyltransferases EZH2 and DOT1L is an effective therapy for neuroblastoma

Janith A Seneviratne; Daenikka Ravindrarajah; Daniel R Carter; Vicki Zhai; Amit Lalwani; Sukriti Krishan; Anushree Balachandran; Ernest Ng; Ruby Pandher; Matthew Wong; Tracy L Nero; Shudong Wang; Murray D Norris; Michelle Haber; Tao Liu; Michael W Parker; Belamy B Cheung; Glenn M Marshall

doi:10.1002/cam4.70082

Combined inhibition of histone methyltransferases EZH2 and DOT1L is an effective therapy for neuroblastoma

Cancer Med. 2024 Nov;13(21):e70082. doi: 10.1002/cam4.70082.

Authors

Janith A Seneviratne^{1

2}, Daenikka Ravindrarajah^{1

2}, Daniel R Carter^{1

3}, Vicki Zhai¹, Amit Lalwani¹, Sukriti Krishan^{1

2}, Anushree Balachandran¹, Ernest Ng^{1

2}, Ruby Pandher^{1

2}, Matthew Wong^{1

2}, Tracy L Nero⁴, Shudong Wang⁵, Murray D Norris^{1

2

6}, Michelle Haber^{1

2}, Tao Liu^{1

2}, Michael W Parker^{4

7}, Belamy B Cheung^{1

2}, Glenn M Marshall^{1

2

8}

Affiliations

¹ Children's Cancer Institute Australia for Medical Research, Lowy Cancer Research Centre, UNSW Sydney, Kensington, New South Wales, Australia.
² School of Clinical Medicine, Faculty of Medicine and Health, UNSW Sydney, Kensington, New South Wales, Australia.
³ School of Biomedical Engineering, University of Technology Sydney, Sydney, New South Wales, Australia.
⁴ Department of Biochemistry and Pharmacology, Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne, Parkville, Victoria, Australia.
⁵ Centre for Drug Discovery and Development, Clinical and Health Sciences, University of South Australia, Adelaide, South Australia, Australia.
⁶ Centre for Childhood Cancer Research, UNSW Sydney, Randwick, New South Wales, Australia.
⁷ ACRF Rational Drug Discovery Centre, St. Vincent's Institute of Medical Research, Fitzroy, Victoria, Australia.
⁸ Kids Cancer Centre, Sydney Children's Hospital, Randwick, New South Wales, Australia.

PMID: 39501501
DOI: 10.1002/cam4.70082

Abstract

Background: The child cancer, neuroblastoma (NB), is characterised by a low incidence of mutations and strong oncogenic embryonal driver signals. Many new targeted epigenetic modifier drugs have failed in human trials as monotherapy.

Methods: We performed a high-throughput, combination chromatin-modifier drug screen against NB cells. We screened 13 drug candidates in 78 unique combinations.

Results: We found that the combination of two histone methyltransferase (HMT) inhibitors: GSK343, targeting EZH2, and SGC0946, targeting DOT1L, demonstrated the strongest synergy across 8 NB cell lines, with low normal fibroblast toxicity. High mRNA expression of both EZH2 and DOT1L in NB tumour samples correlated with the poorest patient survival. Combination HMT inhibitor treatment caused activation of ATF4-mediated endoplasmic reticulum (ER) stress responses. In addition, glutathione and several amino acids were depleted by HMT inhibitor combination on mass spectrometry analysis. The combination of SGC0946 and GSK343 reduced tumour growth in comparison to single agents.

Conclusion: Our results support further investigation of HMT inhibitor combinations as a therapeutic approach in NB.

Keywords: biomarkers; cancer biology; chromatin modifications and dynamics; epigenetics; molecular biology; neuroblastoma.

MeSH terms

Activating Transcription Factor 4 / genetics
Activating Transcription Factor 4 / metabolism
Animals
Antineoplastic Combined Chemotherapy Protocols / pharmacology
Antineoplastic Combined Chemotherapy Protocols / therapeutic use
Cell Line, Tumor
Cell Proliferation / drug effects
Drug Synergism
Endoplasmic Reticulum Stress / drug effects
Enhancer of Zeste Homolog 2 Protein* / antagonists & inhibitors
Enhancer of Zeste Homolog 2 Protein* / genetics
Enhancer of Zeste Homolog 2 Protein* / metabolism
Enzyme Inhibitors / pharmacology
Enzyme Inhibitors / therapeutic use
Histone-Lysine N-Methyltransferase* / antagonists & inhibitors
Histone-Lysine N-Methyltransferase* / genetics
Histone-Lysine N-Methyltransferase* / metabolism
Humans
Indazoles
Mice
Neuroblastoma* / drug therapy
Neuroblastoma* / genetics
Neuroblastoma* / metabolism
Neuroblastoma* / pathology
Pyridones
Xenograft Model Antitumor Assays

Substances

Enhancer of Zeste Homolog 2 Protein
Histone-Lysine N-Methyltransferase
DOT1L protein, human
GSK343
EZH2 protein, human
Activating Transcription Factor 4
ATF4 protein, human
Enzyme Inhibitors
Indazoles
Pyridones

Abstract

MeSH terms

Substances

Grants and funding