Cross-species array comparative genomic hybridization identifies novel oncogenic events in zebrafish and human embryonal rhabdomyosarcoma.
Cross-species array comparative genomic hybridization identifies novel oncogenic events in zebrafish and human embryonal rhabdomyosarcoma.
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DOI：
10.1371/journal.pgen.1003727
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发表时间：
2013-08
期刊：
影响因子：
4.5
通讯作者：
Langenau DM
中科院分区：
文献类型：
--
作者：
Chen EY;Dobrinski KP;Brown KH;Clagg R;Edelman E;Ignatius MS;Chen JY;Brockmann J;Nielsen GP;Ramaswamy S;Keller C;Lee C;Langenau DM
人类癌症基因组高度复杂，这使得确定癌症生长、进展和肿瘤维持的特定驱动因素具有挑战性。为了克服这一障碍，我们将阵列比较基因组杂交（阵列CGH）应用于斑马鱼胚胎横纹肌（ERMS），并利用跨物种比较快速识别人类疾病的基因组拷贝数畸变和新的候选癌基因。斑马鱼的ERMS包含小的，低拷贝扩增的焦点区域。这些相同的区域通常在人类疾病中被放大。例如，在斑马鱼ERMS中鉴定的19个染色体中，有16个在人类疾病中也表现出局灶性的低拷贝增加。在人类和斑马鱼ERMS中，研究人员评估了在扩增基因组区域中发现的基因在促进肿瘤持续生长方面的功能作用，确定了与肿瘤维持相关的关键基因。敲低研究确定了Cyclin D2 （CCND2）、Homeobox Protein C6 （HOXC6）和PlexinA1 （PLXNA1）在人ERMS细胞增殖中的重要作用。PLXNA1敲低也能增强分化、减少迁移和改变非锚定生长。相比之下，血管内皮生长因子（VEGF）信号的化学抑制减少了erms携带斑马鱼的血管生成和肿瘤大小。重要的是，VEGF表达与ERMS患者的不良临床结果相关，这意味着VEGF途径抑制剂是一种有希望改善患者生存的治疗方法。我们的研究结果证明了阵列CGH和跨物种比较在确定人类癌症发病机制所必需的候选癌基因方面的效用。癌症是一种复杂的遗传疾病，通常与基因组DNA片段的区域增益和损失有关。这些变化导致基因表达异常并驱动肿瘤持续生长。由于扩增和删除的DNA片段往往跨越染色体的大片区域，因此确定持续肿瘤生长和进展所需的基因一直具有挑战性。阵列比较基因组杂交技术（Array comparative genomic hybridization， CGH）是鉴别肿瘤基因组拷贝数异常变异的有效技术。在这项研究中，阵列CGH被用于斑马鱼胚胎横纹肌肉瘤（一种小儿肌肉肿瘤）模型。我们的工作表明，斑马鱼癌症基因组中含有少量复发性DNA拷贝数变化，这些变化在人类疾病中也经常被放大。此外，这些染色体区域很小，便于快速鉴定候选癌基因。在斑马鱼阵列CGH中鉴定的一个基因子集被优先用于人类ERMS的功能表征，确定了调节增殖、迁移、分化和新生血管的进化保守途径。我们的研究结果证明了人类和斑马鱼癌症的跨物种阵列CGH比较的广泛实用性，并为识别各种恶性肿瘤中的关键致癌基因提供了急需的发现平台。
Human cancer genomes are highly complex, making it challenging to identify specific drivers of cancer growth, progression, and tumor maintenance. To bypass this obstacle, we have applied array comparative genomic hybridization (array CGH) to zebrafish embryonal rhabdomyosaroma (ERMS) and utilized cross-species comparison to rapidly identify genomic copy number aberrations and novel candidate oncogenes in human disease. Zebrafish ERMS contain small, focal regions of low-copy amplification. These same regions were commonly amplified in human disease. For example, 16 of 19 chromosomal gains identified in zebrafish ERMS also exhibited focal, low-copy gains in human disease. Genes found in amplified genomic regions were assessed for functional roles in promoting continued tumor growth in human and zebrafish ERMS – identifying critical genes associated with tumor maintenance. Knockdown studies identified important roles for Cyclin D2 (CCND2), Homeobox Protein C6 (HOXC6) and PlexinA1 (PLXNA1) in human ERMS cell proliferation. PLXNA1 knockdown also enhanced differentiation, reduced migration, and altered anchorage-independent growth. By contrast, chemical inhibition of vascular endothelial growth factor (VEGF) signaling reduced angiogenesis and tumor size in ERMS-bearing zebrafish. Importantly, VEGFA expression correlated with poor clinical outcome in patients with ERMS, implicating inhibitors of the VEGF pathway as a promising therapy for improving patient survival. Our results demonstrate the utility of array CGH and cross-species comparisons to identify candidate oncogenes essential for the pathogenesis of human cancer. Cancer is a complex genetic disease that is often associated with regional gains and losses of genomic DNA segments. These changes result in aberrant gene expression and drive continued tumor growth. Because amplified and deleted DNA segments tend to span large regions of chromosomes, it has been challenging to identify the genes that are required for continued tumor growth and progression. Array comparative genomic hybridization (array CGH) is an effective technology in identifying abnormal copy number variations in cancer genomes. In this study, array CGH was used in a zebrafish model of embryonal rhabdomyosarcoma - a pediatric muscle tumor. Our work shows that the zebrafish cancer genome contains a small number of recurrent DNA copy number changes, which are also commonly amplified in the human disease. Moreover, these chromosomal regions are small, facilitating rapid identification of candidate oncogenes. A subset of genes identified in zebrafish array CGH was prioritized for functional characterization in human ERMS, identifying evolutionarily conserved pathways that regulate proliferation, migration, differentiation, and neovascularization. Our results demonstrate the broad utility of cross-species array CGH comparisons of human and zebrafish cancer and provide a much needed discovery platform for identifying critical cancer-causing genes in a wide range of malignancies.
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影响因子：
3.7
作者：
Freeman, Jennifer L.;Ceol, Craig;Feng, Hui;Langenau, David M.;Belair, Cassandra;Stern, Howard M.;Song, Anhua;Paw, Barry H.;Look, A. Thomas;Zhou, Yi;Zon, Leonard I.;Lee, Charles
通讯作者：
Lee, Charles
影响因子：
64.5
作者：
Goessling W;North TE;Loewer S;Lord AM;Lee S;Stoick-Cooper CL;Weidinger G;Puder M;Daley GQ;Moon RT;Zon LI
通讯作者：
Zon LI
影响因子：
3.7
作者：
Bridge, JA;Liu, J;Barr, FG
通讯作者：
Barr, FG
影响因子：
4.7
作者：
Davari, Parastoo;Hebert, Jennifer L.;Epstein, Ervin H., Jr.
通讯作者：
Epstein, Ervin H., Jr.
影响因子：
3
作者：
Fountzilas, George;Fragkoulidi, Anna;Marselos, Marios
通讯作者：
Marselos, Marios