Targeted Therapy of TERT-Rearranged Neuroblastoma with BET Bromodomain Inhibitor and Proteasome Inhibitor Combination Therapy

Jingwei Chen; Christopher Nelson; Matthew Wong; Andrew E Tee; Pei Y Liu; Ting La; Jamie I Fletcher; Alvin Kamili; Chelsea Mayoh; Christoph Bartenhagen; Toby N Trahair; Ning Xu; Nisitha Jayatilleke; Marie Wong; Hui Peng; Bernard Atmadibrata; Belamy B Cheung; Qing Lan; Tracy M Bryan; Pieter Mestdagh; Jo Vandesompele; Valerie Combaret; Valentina Boeva; Jenny Y Wang; Isabelle Janoueix-Lerosey; Mark J Cowley; Karen L MacKenzie; Alla Dolnikov; Jinyan Li; Patsie Polly; Glenn M Marshall; Roger R Reddel; Murray D Norris; Michelle Haber; Matthias Fischer; Xu D Zhang; Hilda A Pickett; Tao Liu

doi:10.1158/1078-0432.CCR-20-3044

Targeted Therapy of TERT-Rearranged Neuroblastoma with BET Bromodomain Inhibitor and Proteasome Inhibitor Combination Therapy

Clin Cancer Res. 2021 Mar 1;27(5):1438-1451. doi: 10.1158/1078-0432.CCR-20-3044. Epub 2020 Dec 11.

Authors

Jingwei Chen^{1

2}, Christopher Nelson³, Matthew Wong^{1

2}, Andrew E Tee^{1

2}, Pei Y Liu^{1

2}, Ting La⁴, Jamie I Fletcher^{1

2}, Alvin Kamili^{1

2}, Chelsea Mayoh^{1

2}, Christoph Bartenhagen^{5

6}, Toby N Trahair^{1

2}, Ning Xu^{1

2}, Nisitha Jayatilleke^{1

2}, Marie Wong^{1

2}, Hui Peng⁷, Bernard Atmadibrata¹, Belamy B Cheung^{1

2}, Qing Lan⁸, Tracy M Bryan³, Pieter Mestdagh⁹, Jo Vandesompele⁹, Valerie Combaret¹⁰, Valentina Boeva^{11

12}, Jenny Y Wang^{1

2}, Isabelle Janoueix-Lerosey^{13

14}, Mark J Cowley^{1

2}, Karen L MacKenzie³, Alla Dolnikov^{1

2}, Jinyan Li⁷, Patsie Polly¹⁵, Glenn M Marshall^{1

2

16}, Roger R Reddel³, Murray D Norris^{1

2}, Michelle Haber^{1

2}, Matthias Fischer^{5

6}, Xu D Zhang^{4

17}, Hilda A Pickett³, Tao Liu^{18

2}

Affiliations

¹ Children's Cancer Institute, Randwick, Sydney, Australia.
² School of Women's and Children's Health, University of New South Wales Sydney, Sydney, New South Wales, Australia.
³ Children's Medical Research Institute, Faculty of Medicine and Health, The University of Sydney, Sydney, New South Wales, Australia.
⁴ School of Medicine and Public Health, Priority Research Centre for Cancer Research, University of Newcastle, Callaghan, Australia.
⁵ Department of Experimental Pediatric Oncology, Medical Faculty, University Hospital, University of Cologne, Cologne, Germany.
⁶ Center for Molecular Medicine (CMMC), University of Cologne, Cologne, Germany.
⁷ Advanced Analytics Institute, University of Technology Sydney, Ultimo, Australia.
⁸ Department of Neurosurgery, The Second Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China.
⁹ Center for Medical Genetics Ghent, Ghent University, Ghent, Belgium.
¹⁰ Centre Léon-Bérard, Laboratoire de Recherche Translationnelle, Lyon, France.
¹¹ ETH Zürich, Department of Computer Science, Institute for Machine Learning, Swiss Institute of Bioinformaticsics (SIB), Zurich, Switzerland.
¹² Institut Cochin, Inserm U1016, CNRS UMR 8104, Université Paris Descartes UMR-S1016, Paris, France.
¹³ Institut Curie, Paris Sciences et Lettres Research University, INSERM, U830, Equipe Labellisée Ligue contre le Cancer, Paris, France.
¹⁴ SIREDO: Care, Innovation and Research for Children, Adolescents and Young Adults with Cancer, Institut Curie, Paris, France.
¹⁵ Department of Pathology, School of Medical Sciences, UNSW Sydney, Sydney, New South Wales, Australia.
¹⁶ Kids Cancer Centre, Sydney Children's Hospital, Randwick, New South Wales, Australia.
¹⁷ Translational Research Institute, Henan Provincial People's Hospital, Academy of Medical Science, Zhengzhou University, Zhengzhou, China.
¹⁸ Children's Cancer Institute, Randwick, Sydney, Australia. [email protected].

PMID: 33310889
DOI: 10.1158/1078-0432.CCR-20-3044

Abstract

Purpose: TERT gene rearrangement with transcriptional superenhancers leads to TERT overexpression and neuroblastoma. No targeted therapy is available for clinical trials in patients with TERT-rearranged neuroblastoma.

Experimental design: Anticancer agents exerting the best synergistic anticancer effects with BET bromodomain inhibitors were identified by screening an FDA-approved oncology drug library. The synergistic effects of the BET bromodomain inhibitor OTX015 and the proteasome inhibitor carfilzomib were examined by immunoblot and flow cytometry analysis. The anticancer efficacy of OTX015 and carfilzomib combination therapy was investigated in mice xenografted with TERT-rearranged neuroblastoma cell lines or patient-derived xenograft (PDX) tumor cells, and the role of TERT reduction in the anticancer efficacy was examined through rescue experiments in mice.

Results: The BET bromodomain protein BRD4 promoted TERT-rearranged neuroblastoma cell proliferation through upregulating TERT expression. Screening of an approved oncology drug library identified the proteasome inhibitor carfilzomib as the agent exerting the best synergistic anticancer effects with BET bromodomain inhibitors including OTX015. OTX015 and carfilzomib synergistically reduced TERT protein expression, induced endoplasmic reticulum stress, and induced TERT-rearranged neuroblastoma cell apoptosis which was blocked by TERT overexpression and endoplasmic reticulum stress antagonists. In mice xenografted with TERT-rearranged neuroblastoma cell lines or PDX tumor cells, OTX015 and carfilzomib synergistically blocked TERT expression, induced tumor cell apoptosis, suppressed tumor progression, and improved mouse survival, which was largely reversed by forced TERT overexpression.

Conclusions: OTX015 and carfilzomib combination therapy is likely to be translated into the first clinical trial of a targeted therapy in patients with TERT-rearranged neuroblastoma.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Acetanilides / pharmacology*
Animals
Apoptosis
Cell Cycle Proteins / antagonists & inhibitors*
Cell Proliferation
Drug Therapy, Combination
Female
Gene Rearrangement*
Heterocyclic Compounds, 3-Ring / pharmacology*
Humans
Mice
Mice, Inbred BALB C
Mice, Nude
Molecular Targeted Therapy / methods*
Neuroblastoma / drug therapy*
Neuroblastoma / metabolism
Neuroblastoma / pathology
Oligopeptides / pharmacology*
Proteasome Inhibitors / pharmacology
Telomerase / genetics*
Transcription Factors / antagonists & inhibitors*
Xenograft Model Antitumor Assays

Substances

Acetanilides
BRD4 protein, human
Cell Cycle Proteins
Heterocyclic Compounds, 3-Ring
OTX015
Oligopeptides
Proteasome Inhibitors
Transcription Factors
carfilzomib
TERT protein, human
Telomerase