Pharmacologically difficult targets, such as MYC transcription factors, represent a major challenge in cancer therapy. For the childhood cancer neuroblastoma, amplification of the oncogene MYCN is associated with high-risk disease and poor prognosis. Here, we deployed genome-scale CRISPR-Cas9 screening of MYCN-amplified neuroblastoma and found a preferential dependency on genes encoding the polycomb repressive complex 2 (PRC2) components EZH2, EED, and SUZ12. Genetic and pharmacological suppression of EZH2 inhibited neuroblastoma growth in vitro and in vivo. Moreover, compared with neuroblastomas without MYCN amplification, MYCN-amplified neuroblastomas expressed higher levels of EZH2. ChIP analysis showed that MYCN binds at the EZH2 promoter, thereby directly driving expression. Transcriptomic and epigenetic analysis, as well as genetic rescue experiments, revealed that EZH2 represses neuronal differentiation in neuroblastoma in a PRC2-dependent manner. Moreover, MYCN-amplified and high-risk primary tumors from patients with neuroblastoma exhibited strong repression of EZH2-regulated genes. Additionally, overexpression of IGFBP3, a direct EZH2 target, suppressed neuroblastoma growth in vitro and in vivo. We further observed strong synergy between histone deacetylase inhibitors and EZH2 inhibitors. Together, these observations demonstrate that MYCN upregulates EZH2, leading to inactivation of a tumor suppressor program in neuroblastoma, and support testing EZH2 inhibitors in patients with MYCN-amplified neuroblastoma.

CRISPR-Cas9 screen reveals a MYCN-amplified neuroblastoma dependency on EZH2 / Chen, L.; Alexe, G.; Dharia, N. V.; Ross, L.; Iniguez, A. B.; Conway, A. S.; Wang, E. J.; Veschi, V.; Lam, N.; Qi, J.; Clay Gustafson, W.; Nasholm, N.; Vazquez, F.; Weir, B. A.; Cowley, G. S.; Ali, L. D.; Pantel, S.; Jiang, G.; Harrington, W. F.; Lee, Y.; Goodale, A.; Lubonja, R.; Krill-Burger, J. M.; Meyers, R. M.; Tsherniak, A.; Root, D. E.; Bradner, J. E.; Golub, T. R.; Roberts, C. W. M.; Hahn, W. C.; Weiss, W. A.; Thiele, C. J.; Stegmaier, K.. - In: THE JOURNAL OF CLINICAL INVESTIGATION. - ISSN 0021-9738. - 128:1(2018), pp. 446-462. [10.1172/JCI90793]

CRISPR-Cas9 screen reveals a MYCN-amplified neuroblastoma dependency on EZH2

Veschi V.;
2018

Abstract

Pharmacologically difficult targets, such as MYC transcription factors, represent a major challenge in cancer therapy. For the childhood cancer neuroblastoma, amplification of the oncogene MYCN is associated with high-risk disease and poor prognosis. Here, we deployed genome-scale CRISPR-Cas9 screening of MYCN-amplified neuroblastoma and found a preferential dependency on genes encoding the polycomb repressive complex 2 (PRC2) components EZH2, EED, and SUZ12. Genetic and pharmacological suppression of EZH2 inhibited neuroblastoma growth in vitro and in vivo. Moreover, compared with neuroblastomas without MYCN amplification, MYCN-amplified neuroblastomas expressed higher levels of EZH2. ChIP analysis showed that MYCN binds at the EZH2 promoter, thereby directly driving expression. Transcriptomic and epigenetic analysis, as well as genetic rescue experiments, revealed that EZH2 represses neuronal differentiation in neuroblastoma in a PRC2-dependent manner. Moreover, MYCN-amplified and high-risk primary tumors from patients with neuroblastoma exhibited strong repression of EZH2-regulated genes. Additionally, overexpression of IGFBP3, a direct EZH2 target, suppressed neuroblastoma growth in vitro and in vivo. We further observed strong synergy between histone deacetylase inhibitors and EZH2 inhibitors. Together, these observations demonstrate that MYCN upregulates EZH2, leading to inactivation of a tumor suppressor program in neuroblastoma, and support testing EZH2 inhibitors in patients with MYCN-amplified neuroblastoma.
2018
Epigenetics; Oncology; Cell Line; Tumor; Humans; Neurons; CRISPR-Cas Systems; Cell Differentiation; Enhancer of Zeste Homolog 2 Protein; Gene Amplification; Gene Expression Regulation; Neoplastic; N-Myc Proto-Oncogene Protein; Neuroblastoma; Up-Regulation
01 Pubblicazione su rivista::01a Articolo in rivista
CRISPR-Cas9 screen reveals a MYCN-amplified neuroblastoma dependency on EZH2 / Chen, L.; Alexe, G.; Dharia, N. V.; Ross, L.; Iniguez, A. B.; Conway, A. S.; Wang, E. J.; Veschi, V.; Lam, N.; Qi, J.; Clay Gustafson, W.; Nasholm, N.; Vazquez, F.; Weir, B. A.; Cowley, G. S.; Ali, L. D.; Pantel, S.; Jiang, G.; Harrington, W. F.; Lee, Y.; Goodale, A.; Lubonja, R.; Krill-Burger, J. M.; Meyers, R. M.; Tsherniak, A.; Root, D. E.; Bradner, J. E.; Golub, T. R.; Roberts, C. W. M.; Hahn, W. C.; Weiss, W. A.; Thiele, C. J.; Stegmaier, K.. - In: THE JOURNAL OF CLINICAL INVESTIGATION. - ISSN 0021-9738. - 128:1(2018), pp. 446-462. [10.1172/JCI90793]
File allegati a questo prodotto
Non ci sono file associati a questo prodotto.

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/1698866
 Attenzione

Attenzione! I dati visualizzati non sono stati sottoposti a validazione da parte dell'ateneo

Citazioni
  • ???jsp.display-item.citation.pmc??? 79
  • Scopus 112
  • ???jsp.display-item.citation.isi??? 104
social impact