Sleeping Beauty mutagenesis in a mouse medulloblastoma model defines networks that discriminate between human molecular subgroups
Sleeping Beauty mutagenesis in a mouse medulloblastoma model defines networks that discriminate between human molecular subgroups
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DOI：
10.1073/pnas.1318639110
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发表时间：
2013-11-12
影响因子：
11.1
通讯作者：
Wainwright, Brandon J.
中科院分区：
文献类型：
--
作者：
Genovesi, Laura A.;Ng, Ching Ging;Wainwright, Brandon J.
睡美人（SB）转座子诱变筛选是一个强大的工具，以促进发现癌症基因，驱动肿瘤发生的小鼠模型。在这项研究中，我们试图确定基因的功能与音刺猬信号启动髓母细胞瘤（MB），小脑肿瘤。通过将SB诱变与Patched 1杂合子小鼠（Ptch 1（lacZ/+））相结合，我们观察到与Ptch 1（lacZ/+）对照组相比，MB频率增加，无瘤生存率降低。通过对85个肿瘤的分析，我们确定了77个常见的插入位点，这些位点映射到56个可能导致肿瘤发生增加的基因。将诱变筛选中鉴定的共同插入位点基因定位到人直系同源物，其用于从独立的一组先前描述的人MB样品中选择探针和相应的表达数据，并且令人惊讶地能够准确聚类MB的已知分子亚组，从而定义所有形式的MB的共同调控网络，而不管亚组。我们进行了网络分析，以发现可能的作用机制的子网络，并使用体内模型，以确认一个高度排名的候选基因，Nfia，在促进MB形成的作用。我们的分析表明，候选癌症基因在细胞凋亡和翻译延长的失调，并揭示了一个强大的签名转录调控，将有广泛的影响MB的表达程序。这些网络为人类MB的复杂生物学提供了功能性见解，并确定了所有临床亚组共同干预的潜在途径。
The Sleeping Beauty (SB) transposon mutagenesis screen is a powerful tool to facilitate the discovery of cancer genes that drive tumorigenesis in mouse models. In this study, we sought to identify genes that functionally cooperate with sonic hedgehog signaling to initiate medulloblastoma (MB), a tumor of the cerebellum. By combining SB mutagenesis with Patched1 heterozygous mice (Ptch1(lacZ/+)), we observed an increased frequency of MB and decreased tumor-free survival compared with Ptch1(lacZ/+) controls. From an analysis of 85 tumors, we identified 77 common insertion sites that map to 56 genes potentially driving increased tumorigenesis. The common insertion site genes identified in the mutagenesis screen were mapped to human orthologs, which were used to select probes and corresponding expression data from an independent set of previously described human MB samples, and surprisingly were capable of accurately clustering known molecular subgroups of MB, thereby defining common regulatory networks underlying all forms of MB irrespective of subgroup. We performed a network analysis to discover the likely mechanisms of action of subnetworks and used an in vivo model to confirm a role for a highly ranked candidate gene, Nfia, in promoting MB formation. Our analysis implicates candidate cancer genes in the deregulation of apoptosis and translational elongation, and reveals a strong signature of transcriptional regulation that will have broad impact on expression programs in MB. These networks provide functional insights into the complex biology of human MB and identify potential avenues for intervention common to all clinical subgroups.