TY - JOUR
T1 - CRISPR-Cas9 screen reveals a MYCN-amplified neuroblastoma dependency on EZH2
AU - Veschi, Veronica
AU - Bradner, James E.
AU - Lubonja, Rakela
AU - Veschi, Veronica
AU - Dharia, Neekesh V.
AU - Weir, Barbara A.
AU - Krill-Burger, John M.
AU - Jiang, Guozhi
AU - Root, David E.
AU - Roberts, Charles W.M.
AU - Ross, Linda
AU - Tsherniak, Aviad
AU - Ali, Levi D.
AU - Cowley, Glenn S.
AU - Chen, Liying
AU - Golub, Todd R.
AU - Nasholm, Nicole
AU - Meyers, Robin M.
AU - Hahn, William C.
AU - Clay Gustafson, null
AU - Qi, Jun
AU - Conway, Amy Saur
AU - Lam, Norris
AU - Vazquez, Francisca
AU - Harrington, William F.
AU - Pantel, Sasha
AU - Goodale, Amy
AU - Lee, Yenarae
AU - Iniguez, Amanda Balboni
AU - Wang, Emily Jue
AU - Stegmaier, Kimberly
AU - Alexe, Gabriela
AU - Thiele, Carol J.
AU - Weiss, William A.
AU - Lee, Yenarae
PY - 2018
Y1 - 2018
N2 - 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.
AB - 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.
UR - http://hdl.handle.net/10447/404706
M3 - Article
VL - 128
SP - 446
EP - 462
JO - THE JOURNAL OF CLINICAL INVESTIGATION
JF - THE JOURNAL OF CLINICAL INVESTIGATION
SN - 0021-9738
ER -