The most frequent focal alterations in human retinoblastoma are mutations in the tumor-suppressor gene retinoblastoma (RB) and amplification of the oncogene MYCN. Whether MYCN overexpression drives retinoblastoma has not been assessed in model systems. Here, we have shown that Rb inactivation collaborates strongly with MYCN overexpression and leads to retinoblastoma in mice. Overexpression of human MYCN in the context of Rb inactivation increased the expression of MYC-, E2F-, and ribosome-related gene sets, promoted excessive proliferation, and led to retinoblastoma with anaplastic changes. We then modeled responses to MYCN-directed therapy by suppressing MYCN expression in MYCN-driven retinoblastomas. Initially, MYCN suppression led to proliferation arrest and partial tumor regression with loss of anaplasia. However, over time, retinoblastomas reemerged, typically without reactivation of human MYCN or amplification of murine Mycn. A subset of returning retinoblastomas showed genomic amplification of a Mycn target gene encoding the miR cluster miR-17~92, while most retinoblastomas reemerged without clear genetic alterations in either Mycn or known Mycn targets. This Rb/MYCN model recapitulates key genetic driver alterations seen in human retinoblastoma and reveals the emergence of MYCN independence in an initially MYCN-driven tumor.
Authors
Nan Wu, Deshui Jia, Breanna Bates, Ryan Basom, Charles G. Eberhart, David MacPherson
(A) GSEA comparing Rb-null (n = 4) with Rb/MYCN (n = 4) retinae at P12. The 50-gene “Hallmark signatures” set from MSigDB was queried and showed MYC- and E2F-related gene sets as the top 2 most significant gene sets enriched upon MYCN overexpression in the Rb-null retina. (B) Top 10 gene sets enriched in Rb/TET-MYCN and top 3 gene sets in Rb-null P12 retinae, with the normalized enrichment score (NES) shown. (C) Enriched GO terms in genes upregulated in Rb/TET-MYCN relative to Rb-null P12 retinae. (D) Enriched GO terms in genes upregulated in Rb-null relative to Rb/TET-MYCN P12 retinae. MF, molecular function; BP, biological process; CC, cellular component.