Sleeping Beauty transposon mutagenesis identifies genes that cooperate with mutant Smad4 in gastric cancer development
Sleeping Beauty transposon mutagenesis identifies genes that cooperate with mutant Smad4 in gastric cancer development
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DOI：
10.1073/pnas.1603223113
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发表时间：
2016-04-05
影响因子：
11.1
通讯作者：
Copeland, Neal G.
中科院分区：
文献类型：
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作者：
Takeda, Haruna;Rust, Alistair G.;Copeland, Neal G.
Smad4的突变容易导致胃肠道癌症，这是与癌症相关的死亡的第三大原因。为了确定胃癌(GC)发生的驱动基因，我们在Smad4(+/-)突变小鼠的胃中进行了睡美人(SB)转座子突变筛选。这一筛选确定了59个候选GC干线司机和更多数量的候选GC进展基因。引人注目的是，22个SB鉴定的干线司机是已知或候选的癌症基因，而四个SB鉴定的干线司机，包括PTEN，Smad4，RNF43和NF1，是已知的人类GC干线司机。与人类GC相似，通路分析发现WNT、转化生长因子-β和PI3K-PTEN信号、泛素介导的蛋白分解、粘连连接和RNA降解以及与染色质修饰和组织有关的基因是GC中高度非调控的通路。对完整的SB鉴定基因进行比较癌基因组筛选，发现它们高度富含在人类GC中突变的基因，并鉴定出许多候选的人类GC基因。最后，通过将我们的SB识别基因的完整列表与五个大规模人类GC测序研究中发现的突变基因列表进行比较，我们确定了低密度脂蛋白受体相关蛋白1B(LRP1B)是一个先前未确定的人类候选GC肿瘤抑制基因。在测序的462个人类GC样本中，LRP1B发现了129个突变，LRP1B是3312个人类癌症中发现的前十大缺失基因之一。因此，SB突变有助于分类驱动Smad4诱导的GC生长的合作分子机制，并发现在人类GC中具有潜在临床意义的基因。
Mutations in SMAD4 predispose to the development of gastrointestinal cancer, which is the third leading cause of cancer-related deaths. To identify genes driving gastric cancer (GC) development, we performed a Sleeping Beauty (SB) transposon mutagenesis screen in the stomach of Smad4(+/-) mutant mice. This screen identified 59 candidate GC trunk drivers and a much larger number of candidate GC progression genes. Strikingly, 22 SB-identified trunk drivers are known or candidate cancer genes, whereas four SB-identified trunk drivers, including PTEN, SMAD4, RNF43, and NF1, are known human GC trunk drivers. Similar to human GC, pathway analyses identified WNT, TGF-beta, and PI3K-PTEN signaling, ubiquitin-mediated proteolysis, adherens junctions, and RNA degradation in addition to genes involved in chromatin modification and organization as highly deregulated pathways in GC. Comparative oncogenomic filtering of the complete list of SB-identified genes showed that they are highly enriched for genes mutated in human GC and identified many candidate human GC genes. Finally, by comparing our complete list of SB-identified genes against the list of mutated genes identified in five large-scale human GC sequencing studies, we identified LDL receptor-related protein 1B (LRP1B) as a previously unidentified human candidate GC tumor suppressor gene. In LRP1B, 129 mutations were found in 462 human GC samples sequenced, and LRP1B is one of the top 10 most deleted genes identified in a panel of 3,312 human cancers. SB mutagenesis has, thus, helped to catalog the cooperative molecular mechanisms driving SMAD4-induced GC growth and discover genes with potential clinical importance in human GC.