Murine & Human In Vivo Models of Melanoma Formation

鼠类

• 批准号: 7246102
• 负责人: NORMAN E SHARPLESS
• 金额: $ 33.37万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7246102
• 关键词: Accounting; Address; Alleles; Biological Models; CDKN2A gene; Characteristics; DNA Damage; Databases; Dermal; Disease Progression; Dose; Event; Exposure to; Extracellular Signal Regulated Kinases; Formalin; Genetic; Genetic Models; Genetically Engineered Mouse; Growth; Heterogeneity; Human; Human Genetics; Immunodeficient Mouse; Kinetics; Lesion; Link; Modeling; Molecular; Mus; Mutation; NRAS gene; Neonatal; Neoplasm Metastasis; Neoplasms; Nevus; Oncogenic; Outcome; Paraffin Embedding; Primary Neoplasm; Reagent; Skin; TP53 gene; UV Radiation Exposure; UV induced; Work; in vivo; in vivo Model; melanocyte; melanoma; novel; response; senescence; tumor

Project 3 utilizes human and murine model systems to address the links between signature genetic events in melanoma progression (p16INK4a loss and N-RAS/B-RAF mutation) and the DMAdamage response in melanocytes. This project combines analyses of novel murine models of melanoma with a comprehensive molecular and immunohistochemical study of a large, clinically annotated human melanoma database. In specific aim 1, we combine two different RAS alleles, conditionally inactivatable p53 and p16INK4a alleles, and an inducible, melanocyte-specific CRE allele to produce new murine models of melanoma that are highly faithful to the human genetics of this tumor. In these models, all oncogenic events are restricted to the melanocytic compartment, and include somatic inactivation of p16/p53 or somatic activation of K-RAS, Two of the alleles (Tyr-CRE-ER-T and conditional p16INK4a) are newly characterized and unpublished. In specific aim 2, we combine these novel murine models of melanoma with neonatal UV-B exposure to facilitate in vivo melanomagenesis. Specific aim 2 also includes a detailed immunohistochemical analysis of the kinetics of the expression of markers of the DNA damage response and senescence with or without UV- B treatment in the setting of RAS activation, p16INK4a loss and/or p53 loss. This specific aim employs a similar approach to analyze human dermal reconstructs in immunodeficient mice with and without UV exposure using reagents supplied from projects 1 and 2. In specific aim 3, we extend our analysis of human primary formalin-fixed and paraffin-embedded melanocytic lesions to identify the relationship among RAS/RAF/p16INK4a mutation, ERK MAP kinase activation, senescence and the DNA damage response in the progression from nevus to metastatic tumor. This specific aim includes a comprehensive immunohistochemical analysis of several markers of the DNA damage response and senescence, as well as a mutational analysis of N-RAS, B-RAF and p16INK4a. This specific aim is powered (250 melanocytic lesions from nevus to metaststic melanoma) to account for molecular heterogeneity in primary tumors but still uncover biologically significant relationships among these signature genetic events, the DNA damage response and melanoma progression. We expect this work will further our basic understanding of human melanoma progression as well as identify new clinjcal predictors of disease progression and outcome.

项目3利用人类和鼠模型系统来解决签名遗传事件之间的联系 黑色素瘤进展（P16INK4A损失和N-RAS/B-RAF突变）和DMADAMAGE反应 黑素细胞。该项目结合了新型黑色素瘤鼠模型的分析和全面 大型临床注释的人黑色素瘤数据库的分子和免疫组织化学研究。在 特定的目标1，我们将两个不同的RAS等位基因结合在一起，有条件地是不可分割的p53和P16INK4A等位基因， 以及可诱导的，黑色素细胞特异性的CRE等位基因，以产生新的黑色素瘤模型 非常忠于这种肿瘤的人类遗传学。在这些模型中，所有致癌事件都仅限于 黑素细胞室，包括p16/p53的躯体灭活或k-ras的体细胞激活，两个 等位基因（Tyr-Cre-er-T和条件P16INK4A）的新特征和未发表。在 特定的目标2，我们将这些新型黑色素瘤的新鼠模型与新生儿UV-B暴露于 促进体内黑色素敏作。特定目标2还包括对的详细免疫组织化学分析 DNA损伤反应和衰老的标记表达的动力学，有或没有紫外线 B治疗RAS激活，P16INK4A丢失和/或p53损失。这个特定目标采用 类似的方法来分析有或没有紫外线的免疫缺陷小鼠中人类皮肤重建 使用项目1和2提供的试剂暴露。在特定目标3中，我们扩展了对人类的分析 原发性福尔马林固定和石蜡包裹的黑素细胞病变，以识别 RAS/RAF/P16INK4A突变，ERK MAP激酶激活，衰老和DNA损伤反应 从奈夫到转移性肿瘤的进展。这个具体目的包括一个全面的 DNA损伤反应和衰老的几个标记的免疫组织化学分析以及 N-RAS，B-RAF和P16INK4A的突变分析。该特定目的是动力的（250个黑色素细胞 从颈部到转移性黑色素瘤的病变）考虑了原发性肿瘤的分子异质性，但 在这些标志性遗传事件中仍然发现生物学上具有重要意义的关系，DNA损害 反应和黑色素瘤进展。我们希望这项工作将进一步我们对人类的基本理解 黑色素瘤的进展以及鉴定疾病进展和预后的新临床预测指标。