文章：

在小鼠中进行高通量插入突变筛选以确定致癌网络

High-throughput insertional mutagenesis screens in mice to identify oncogenic networks

原文发布日期：2009-06-01

DOI: 10.1038/nrc2647

类型: Review Article

开放获取: 否

要点：

1. The integration of slow transforming retroviruses into the genome of host cells can lead to deregulation of nearby genes. Cells can be infected repeatedly, acquiring multiple mutations that can lead to the development of a tumour.
2. Genes implicated in tumour formation are found through the identification of proviral or transposon insertions in independent tumours at frequencies higher than would be expected by chance. Genes in loci that recurrently contain proviral or transposon insertions in independent tumours show a significant overlap with known human oncogenes and tumour suppressor genes.
3. The arrival of next-generation sequencing technologies allows identification of most if not all insertions in a tumour. This will enable a more comprehensive identification of cancer genes and cooperating pairs of cancer genes and facilitate the establishment of the functional role of these cancer genes in tumour development.
4. Examining the genes that collaborate with common mutations in the Myc, p53, Rb and Ras pathways may identify genes that are essential for activating or abrogating the tumorigenic effects of these mutations. Using retrovirus- or transposon-mediated insertional mutagenesis to identify genes that collaborate with genes of the Myc, p53, Rb and Ras pathways may expand the range of targets for developing novel cancer therapeutics.
5. Retrovirus- or transposon-mediated insertional mutagenesis screens can be used to identify genes that confer resistance to drugs both in vitro and in vivo and may thus provide pivotal information to improve existing therapeutic strategies.
6. Although most proviral insertions are activating rather than inactivating, a substantial fraction of the insertions are found to abrogate gene function, thus enabling the identification of tumour suppressor genes.
7. Cross-comparison of insertional mutagenesis data from mice with mutation data from human tumour panels may help identify the 'driver' mutations in human cancer. With the development of insertional mutagenesis strategies for solid tissues these cross-comparisons will become even more powerful, as they will allow direct comparison of human tumour data with insertions derived from their cognate mouse tumours.
8. Scrutinizing concurrent and mutually exclusive insertions in clonal cell populations of tumours or in single cells of a tumour in combination with high-quality expression profiling and proteomics analysis may teach us about underlying mechanisms of tumour heterogeneity and their role in tumour initiation and maintenance.

要点翻译：

1. 慢转化逆转录病毒整合入宿主细胞基因组可能导致邻近基因的失调。细胞可被反复感染，积累多重突变，最终诱发肿瘤形成。
2. 通过鉴定独立肿瘤中前病毒或转座子插入频率显著高于随机预期的现象，可发现与肿瘤形成相关的基因。在独立肿瘤中反复出现前病毒或转座子插入的基因位点，与已知人类癌基因和抑癌基因存在显著重叠。
3. 新一代测序技术的出现使得鉴定肿瘤中全部（或近乎全部）插入位点成为可能。这将有助于更全面地识别癌基因及其协同作用组合，并促进明确这些癌基因在肿瘤发展中的功能作用。
4. 研究与Myc、p53、Rb和Ras通路常见突变协同作用的基因，或可揭示激活或阻断这些突变致瘤效应的关键基因。利用逆转录病毒或转座子介导的插入突变筛选与Myc、p53、Rb和Ras通路基因协同作用的基因，有望拓展新型癌症治疗靶点的范围。
5. 逆转录病毒或转座子介导的插入突变筛选可用于识别体外和体内赋予耐药性的基因，从而为改进现有治疗策略提供关键信息。
6. 虽然大多数前病毒插入呈现激活而非失活效应，但相当比例的插入确实会破坏基因功能，这使得抑癌基因的识别成为可能。
7. 对小鼠插入突变数据与人类肿瘤基因突变数据进行交叉比对，有助于识别人类癌症中的"驱动"突变。随着实体组织插入突变策略的发展，这类交叉比对将更具价值，因其可实现人类肿瘤数据与对应小鼠肿瘤插入突变的直接比较。
8. 通过结合高质量表达谱和蛋白质组学分析，深入探究肿瘤克隆细胞群或单细胞中并发及互斥的插入事件，可揭示肿瘤异质性的潜在机制及其在肿瘤发生和维持中的作用。

英文摘要：

Retroviral insertional mutagenesis screens have been used for many years as a tool for cancer gene discovery. In recent years, completion of the mouse genome sequence as well as improved technologies for cloning and sequencing of retroviral insertions have greatly facilitated the retrieval of more complete data sets from these screens. The concomitant increase of the size of the screens allows researchers to address new questions about the genes and signalling networks involved in tumour development. In addition, the development of new insertional mutagenesis tools such as DNA transposons enables screens for cancer genes in tissues that previously could not be analysed by retroviral insertional mutagenesis.

摘要翻译： 

逆转录病毒插入突变筛选多年来一直被用作癌症基因发现的工具。近年来，随着小鼠基因组序列的完成以及克隆和测序逆转录病毒插入片段技术的改进，从这些筛选中获取更完整数据集的能力大大增强。筛选规模的同步扩大使研究人员能够就肿瘤发生所涉及的基因和信号网络提出新问题。此外，DNA转座子等新型插入突变工具的开发，使得以往无法通过逆转录病毒插入突变进行分析的组织也得以用于癌症基因筛选。

原文链接：

High-throughput insertional mutagenesis screens in mice to identify oncogenic networks