Identification of Tumor Promotion Susceptibility Genes

促癌易感基因的鉴定

• 批准号: 8367236
• 负责人: John DiGiovanni
• 金额: $ 32.85万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-12-18 至 2014-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8367236
• 关键词: Acetates; Acetone; Affect; Animal Model; Animals; Applications Grants; C57BL/6 Mouse; Chromosome Mapping; Chromosomes, Human, Pair 1; Chromosomes, Human, Pair 9; Congenic Mice; Congenic Strain; DBA/2 Mouse; Data; Development; Differentiation and Growth; Disease; Environmental Carcinogens; Epidemiology; Epidermis; Epithelial; Gene Expression; Gene-Modified; General Population; Generations; Genes; Genetic; Genetic Polymorphism; Glutathione S-Transferase; Goals; Haplotypes; Human; Inherited; Knock-in Mouse; Lead; Link; Lipid Peroxidation; Location; Luciferases; Malignant Neoplasms; Maps; Measures; Methodology; Modeling; Mouse Strains; Mus; Nature; Nucleic Acid Regulatory Sequences; Oxidative Stress; Phorbol Esters; Population; Predisposition; Prevention strategy; Promoter Regions; Protocols documentation; Publishing; Reporter; Research; Resistance; Role; Sequence Analysis; Severities; Skin; Skin Carcinogenesis; Skin Neoplasms; Staging; Study Section; Susceptibility Gene; Syndrome; System; Testing; Tetradecanoylphorbol Acetate; Transgenic Organisms; Tumor Promotion; Variant; Wild Type Mouse; cDNA Arrays; cancer prevention; carcinogenesis; congenic; glutathione S-transferase alpha; interest; keratinocyte; novel; promoter; response; trait; tumor; tumorigenesis

PROJECT SUMMARY/ABSTRACT The primary goal of the research proposed in this grant application is to identify and characterize genes that modify susceptibility to skin tumor promotion using the multi-stage skin carcinogenesis model in mice. Much data from both human epidemiologic and animal studies support the hypothesis that cancer susceptibility in the general population is a function of multiple, poorly penetrant modifier genes that control the propensity toward environmental carcinogen-induced tumor development. Data suggest that susceptibility to the tumor promotion stage is a major determinant of overall susceptibility to multi-stage, epidermal carcinogenesis in mice. Identifying and characterizing genes that modify susceptibility to tumor promotion is crucial for a complete understanding of multistage carcinogenesis and for developing effective cancer prevention strategies. Genes that modify susceptibility and severity of other disease syndromes in humans have successfully been identified in animal models using approaches similar to those proposed in this application to identify and characterize promotion susceptibility loci. To identify genes that modify promotion susceptibility, we have used the classic multi-stage skin tumorigenesis model in the mouse, which is an excellent animal model to study epithelial carcinogenesis in humans. Genetic control of susceptibility to skin tumor promotion by the phorbol ester, 12- O-tetradecanoylphorbol-13-acetate (TPA), in crosses between susceptible DBA/2 and resistant C57BL/6 mice is a multigenic trait and we have mapped promotion susceptibility loci to chromosomes (chr) 1 (Psl3), 2 (Psl2), 9 (Psl1), and 19 (Psl4). Analysis of C57BL/6.Psl1dba congenic mouse strains suggests that at least three genes underlie the effects of Psl1 on skin tumor promotion susceptibility. We have designated these loci as Psl1.1, Psl1.2, and Psl1.3. Furthermore, global gene expression analyses using cDNA microarrays revealed that glutathione S-transferase alpha 4 (Gsta4), which maps within Psl1.2, is expressed at 20-fold higher levels in the epidermis of TPA-treated C57BL/6 compared to DBA/2 mice. Gsta4 is a glutathione-S-transferase (GST) and a major substrate for Gsta4 is 4-hydroxy-2(E)-nonenal (4-HNE), a product of lipid peroxidation. Recent preliminary studies indicate that C57BL/6 mice, null for Gsta4, are more sensitive than wild-type mice to the multi-stage skin tumor protocol, using TPA as a promoter. These observations, taken together with published data supporting a role for Gsta4 in response to oxidative stress, suggests that Gsta4 is a good candidate for a gene that underlies the effect of Psl1.2 on TPA promotion susceptibility. In the proposed research, we will test the hypotheses that Gsta4 modifies the response to TPA skin tumor promotion in DBA/2 and C57BL/6 mice by regulating the level of 4-HNE following TPA treatment. We will further characterize the Psl1.2 locus to determine if any other genes mapping within the locus are modifiers of TPA skin tumor promotion susceptibility. In addition, we will identify and characterize genes mapping to Psl1.1 and Psl1.3 that are associated with responsiveness to TPA-induced skin tumor promotion in DBA/2 and C57BL/6 mice. The Specific Aims are: i) To further characterize the role of Gsta4 as a modifier of TPA skin tumor promotion susceptibility; ii) To characterize the mechanism for, and consequences of, strain-specific induction of Gsta4; and iii) To identify and characterize genes mapping within the Psl1.1 and Psl1.3 loci that modify the response to TPA skin tumor promotion.

项目摘要/摘要 本赠款应用程序提出的研究的主要目标是识别和表征基因 使用小鼠中多阶段的皮肤致癌模型来修改对皮肤肿瘤促进的敏感性。很多 来自人类流行病学和动物研究的数据支持以下假设。 一般人群是多个渗透修饰剂基因的函数，该基因控制着倾向的倾向 环境致癌诱导的肿瘤发育。数据表明对肿瘤促进的敏感性 阶段是对小鼠多阶段表皮癌变的总体敏感性的主要决定因素。 识别和表征改变对肿瘤促进易感性的基因，对于完整的基因至关重要 了解多阶段癌变和制定有效的癌症预防策略。基因 已经成功鉴定出了其他疾病综合症的敏感性和严重程度 在动物模型中，使用与本应用程序中提出的方法相似的方法来识别和表征 促进易感基因座。为了识别改变促进敏感性的基因，我们使用了经典 小鼠中的多阶段皮肤肿瘤发生模型，这是研究上皮的出色动物模型 人类的致癌作用。佛波尔酯对促进皮肤肿瘤的易感性的遗传控制，12-- O-tetradecanoylphorbol-13-乙酸酯（TPA），在易感DBA/2与耐药C57BL/6小鼠之间 是一种多基因性状，我们已将促进敏感性基因座映射到染色体（CHR）1（PSL3），2（PSL2）， 9（PSL1）和19（PSL4）。 C57BL/6.PSL1DBA的Encynic小鼠菌株的分析表明至少三个 基因是PSL1对皮肤肿瘤促进易感性的影响。我们将这些基因座指定为 PSL1.1，PSL1.2和PSL1.3。此外，使用cDNA微阵列的全局基因表达分析显示 谷胱甘肽S-转移酶Alpha 4（GSTA4）在PSL1.2中映射，以20倍高度表示 与DBA/2小鼠相比，在TPA处理的C57BL/6的表皮中。 GSTA4是谷胱甘肽-S-转移酶（GST） GSTA4的主要底物是4-羟基-2（E） - 硝酸（4-HNE），这是脂质过氧化的产物。最近的 初步研究表明，c57bl/6小鼠的gSTA4无效，比野生型小鼠更敏感 多阶段的皮肤肿瘤方案，使用TPA作为启动子。这些观察结果，以及已出版的 支持GSTA4在响应氧化应激方面的作用的数据表明，GSTA4是A的良好候选者 PSL1.2对TPA促进敏感性的影响的基因是基因。在拟议的研究中，我们将测试 GSTA4通过DBA/2和C57BL/6小鼠的TPA皮肤肿瘤促进的反应的假设通过 调节TPA治疗后4-HNE的水平。我们将进一步将PSL1.2基因座描述为 确定该基因座中的任何其他基因映射是否是TPA皮肤肿瘤促进的修饰符 敏感性。此外，我们将识别并表征映射到PSL1.1和PSL1.3的基因 与DBA/2和C57BL/6小鼠中TPA诱导的皮肤肿瘤促进的反应性有关。这 具体目的是：i）进一步表征GSTA4作为TPA皮肤肿瘤促进的修饰符的作用 敏感性； ii）表征GSTA4应变特异性诱导的机制和后果； iii）识别和表征psl1.1和psl1.3基因座的基因映射，以修改响应 促进TPA皮肤肿瘤。

项目成果

A high-throughput 1,536-well luminescence assay for glutathione S-transferase activity.

谷胱甘肽 S-转移酶活性的高通量 1,536 孔发光测定。

• DOI: 10.1089/adt.2009.0248
• 发表时间: 2010
• 期刊: Assay and drug development technologies
• 影响因子: 1.8
• 作者: Yasgar,Adam;Shultz,John;Zhou,Wenhui;Wang,Hui;Huang,Fen;Murphy,Nancy;Abel,ErikaL;DiGiovanni,John;Inglese,James;Simeonov,Anton
• 通讯作者: Simeonov,Anton