Genetic and molecular search for targets of the Tribbles adaptor protein

Tribbles 接头蛋白靶标的遗传和分子搜索

• 批准号: 8953244
• 负责人: LEONARD L DOBENS
• 金额: $ 19.71万
• 依托单位: UNIVERSITY OF MISSOURI KANSAS CITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-06-01 至 2017-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8953244
• 关键词: Adaptor Signaling Protein; Algorithms; Animal Model; Animals; Antineoplastic Agents; Attention; Binding; Cell Death; Cell Proliferation; Cell Size; Cell division; Cells; Chemicals; Collection; Colon Carcinoma; Computers; Core Facility; Data; Defect; Development; Diabetes Mellitus; Diagnosis; Diet; Diet and Nutrition; Disease; Dissection; Drosophila genus; Energy Metabolism; Enzymes; Epidemiologic Studies; Exhibits; Family; Family member; Fee-for-Service Plans; Genes; Genetic; Genetic Models; Genetic Screening; Genetic study; Genomics; Glucose; Goals; Growth; Health; Homeostasis; Human; Hyperinsulinism; Immunoprecipitation; Individual; Insulin; Insulin Receptor; Insulin Resistance; Insulin Signaling Pathway; Insulin-Like Growth Factor I; Knockout Mice; Lead; Lesion; Letters; Libraries; Link; Lipids; Malignant Neoplasms; Malignant neoplasm of thyroid; Mass Spectrum Analysis; Mediating; Mediator of activation protein; Meta-Analysis; Metabolic; Metabolic Diseases; Metabolism; Mitosis; Modeling; Molecular; Molecular Genetics; Mus; Non-Insulin-Dependent Diabetes Mellitus; Obesity; Open Reading Frames; Organism; Outcome; Output; Pathway interactions; Phenotype; Phosphorylation; Phosphotransferases; Population; Positioning Attribute; Precipitation; Predisposition; Premalignant; Prevalence; Protein Family; Protein Isoforms; Proteins; Publishing; Rattus; Reading; Reporting; Role; Signal Pathway; Signal Transduction; Somatomedins; Source; Stress; System; Testing; Time; Tissues; Transgenes; Transgenic Organisms; Up-Regulation; Variant; Whole Organism; Wing; base; cancer cell; cancer type; cdc25 Phosphatase; cell growth; cell motility; diabetic; drug candidate; fly; gene function; genetic regulatory protein; high risk; high throughput screening; improved; insight; insulin mediators; insulin signaling; malignant breast neoplasm; member; mortality; novel; open source; programs; public health relevance; response; tool; transcription factor; tumor; tumor growth; tumor progression

 DESCRIPTION (provided by applicant): Defects in insulin signaling have impacts on both metabolic disease and cancer. Epidemiological studies confirm long time observations that obese individuals and type 2 diabetics with hyperinsulinimia are at higher risk for various types of cancers, but the link between these disorders are unclear. Hyperinsulinimia may directly increase growth of tumor with high levels of insulin receptor, or indirectly increase Insulin-like Growth Factor (IGF) levels, which activates a pathway with well-established links to colon, breast and thyroid cancers. The prevalence of Type 2 diabetes (T2D) is increasing rapidly in populations worldwide, but public programs to improve diet and nutrition have largely failed to reduce the problem and an urgent need exists to understand how the pathway is modulated during cancer progression. Trib family members have been linked to both metabolic defects and cancer formation. This family of proteins shares a conserved kinase line central domain and functions by binding key regulatory proteins - with targets that include transcription factors, kinases and enzymes - to link multiple cell signaling pathways regulating cell growth, proliferation and differentiation. In humans, Tribbles 3 (Trib3) levels are aberrantly high in sufferers of both insulin resistance and T2DM, and a Trib3 protein variant (Trib3Q/R85) has been linked to populations predisposed to metabolic disease. Several anticancer agents have been shown to promote cancer cell death via TRIB3 upregulation and a recent report shows that Trib3 knockout mice exhibit accelerated formation of premalignant lesions due to increased activity of the insulin signaling pathway. Model genetic organisms offer a powerful system to uncover the conserved genetic mechanisms that connect the insulin signaling to cell growth and tissue homeostasis and over the past fifteen years, Drosophila has been used to study the genetics of insulin-regulated metabolism and energy homeostasis. In Drosophila, Tribbles binds and block cdc25 phosphatase, a key trigger of cell proliferation and our recently published data shows that Trbl antagonizes insulin signaling by binding and inhibiting the activation of Akt kinase, a key mediator of the insulin response. Our identification of fly Trbl as a antagonist of insulin signaling puts us in the advantageous position to use the genetic tools available in this simple model organism to identify interacting genes, pathways and drug candidates that as well as the opportunity to carry out rapid dissection of genetic mechanisms, neither of which is feasible in higher model organisms. Here we propose to search for novel Trbl pathway components - including binding partners and pathway targets - combining a genetic screen with a molecular screens. This proposal also develops a computer-based tool which will serve as (1) a new framework for quantitative analysis of cell growth and proliferation, and (2) a versatile, modular, open-source toolbox of algorithms enabling the discovery of genetic pathways, chemical probes, and drug candidates via high-throughput screens in the whole organism with relevance to a variety of diseases. Our goals are to use this simpler model genetic system to identify new conserved Trbl pathway components regulating cell growth and proliferation.

 描述（由申请人提供）：胰岛素信号传导缺陷对代谢疾病和癌症都有影响。流行病学研究证实，长期观察结果表明，肥胖个体和患有高胰岛素血症的 2 型糖尿病患者患各种类型癌症的风险较高，但这些疾病之间的联系尚不清楚。高胰岛素血症可能会直接增加高水平胰岛素受体的肿瘤生长，或间接增加胰岛素样生长因子（IGF）水平，从而激活与结肠癌、乳腺癌和甲状腺癌密切相关的途径。 2 型糖尿病 (T2D) 的患病率在全球人口中迅速增加，但改善饮食和营养的公共计划在很大程度上未能减少这一问题，迫切需要了解在癌症进展过程中如何调节该途径。部落家族成员与代谢缺陷和癌症形成有关。该蛋白家族共享一个保守的激酶系中心结构域，并通过与关键调节蛋白（包括转录因子、激酶和酶等靶标）结合来发挥作用，从而连接调节细胞生长、增殖和分化的多个细胞信号传导途径。在人类中，胰岛素抵抗和 T2DM 患者的 Tribbles 3 (Trib3) 水平异常高，并且 Trib3 蛋白变体 (Trib3Q/R85) 与易患代谢疾病的人群有关。多种抗癌药物已被证明可通过 TRIB3 上调促进癌细胞死亡，最近的一份报告显示，由于胰岛素信号通路活性增加，Trib3 敲除小鼠表现出癌前病变加速形成。模型遗传生物提供了一个强大的系统来揭示将胰岛素信号传导与细胞生长和组织稳态联系起来的保守遗传机制，在过去的十五年中，果蝇已被用来研究胰岛素调节的代谢和能量稳态的遗传学。在果蝇中，Tribbles 结合并阻断 cdc25 磷酸酶，这是细胞增殖的关键触发因素，我们最近发表的数据表明，Trbl 通过结合和抑制 Akt 激酶（胰岛素反应的关键介质）的激活来拮抗胰岛素信号传导。我们将果蝇 Trbl 鉴定为胰岛素信号传导拮抗剂，使我们处于有利地位，可以使用这种简单模型生物中可用的遗传工具来识别相互作用的基因、途径和候选药物，并有机会快速剖析遗传机制，而这两种方法在高等模型生物中都不可行。在这里，我们建议将遗传筛选与分子筛选相结合，寻找新的 Trbl 途径成分 - 包括结合伙伴和途径靶标。该提案还开发了一种基于计算机的工具，该工具将充当（1）细胞生长和增殖定量分析的新框架，以及（2）多功能、模块化、开源算法工具箱，能够通过整个生物体中与多种疾病相关的高通量筛选来发现遗传途径、化学探针和候选药物。我们的目标是使用这个更简单的模型遗传系统来识别调节细胞生长和增殖的新保守 Trbl 通路成分。