High Throughput Screen to Identify Inhibitors of the Golgi GOLPH3 Pathway

用于鉴定高尔基体 GOLPH3 通路抑制剂的高通量筛选

• 批准号: 9006373
• 负责人: SETH J FIELD
• 金额: $ 42.32万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-01 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9006373
• 关键词: Actins; Appearance; Automobile Driving; Binding; Binding Proteins; Biological Assay; Biological Models; Cancer Patient; Cataloging; Catalogs; Cause of Death; Cell Culture Techniques; Cell Death; Cell membrane; Chemicals; Chin; Collaborations; Cytoskeleton; DNA Damage; Data; Development; Drug Kinetics; Electron Microscopy; Excretory function; F-Actin; Frequencies; Genes; Genomics; Goals; Golgi Apparatus; HRAS gene; Human; Institutes; Lead; Link; Literature; Lung; Malignant Neoplasms; Malignant neoplasm of lung; Membrane; Membrane Protein Traffic; Metabolism; Modeling; Molecular Bank; Molecular Target; Myosin ATPase; Oncogenes; Oncogenic; Pathway interactions; Patients; Phosphatidylinositols; Process; Property; Resistance; Role; Shapes; Stratification; Structure-Activity Relationship; Testing; Therapeutic; Therapeutic Agents; Travel; United States National Institutes of Health; Vesicle; Xenograft Model; Xenograft procedure; Yeasts; absorption; actin 2; cancer cell; high throughput screening; inhibitor/antagonist; insight; interest; killings; light microscopy; malignant breast neoplasm; mouse model; new therapeutic target; novel; novel strategies; novel therapeutics; outcome forecast; overexpression; repository; response; screening; small molecule; small molecule inhibitor; tool; trafficking; tumorigenesis

High Throughput Screen to Identify Inhibitors of the Golgi GOLPH3 Pathway PROJECT SUMMARY GOLPH3 is the first example of an oncogene that functions in trafficking from the Golgi to the plasma membrane. The GOLPH3 gene is frequently amplified in human cancers (e.g. 56% of lung cancers) and overexpression of GOLPH3 drives oncogenic transformation in cell culture and xenograft models of cancer. Furthermore, overexpression of GOLPH3 occurs at high frequency in a wide spectrum of cancers and high expression predicts poor patient prognosis. We discovered GOLPH3 as a novel effector of the phosphoinositide PI4P. We showed that GOLPH3 localizes to the Golgi by binding to PI4P. GOLPH3 also binds to an unconventional myosin, MYO18A, which links the Golgi to the actin cytoskeleton. The PI4P/GOLPH3/MYO18A/F-actin pathway functions in Golgi-to-plasma membrane trafficking by applying a tensile force to assist in vesicle budding from the Golgi. This tensile force also helps to give the Golgi its familiar appearance by light and electron microscopy. Thus, the shape of the Golgi is a consequence of the mechanism of trafficking. The ability of GOLPH3 to drive oncogenic transformation in model systems is dependent on the PI4P/GOLPH3/MYO18A/F-actin pathway. Furthermore, we have shown that this GOLPH3 pathway can confer resistance to killing by DNA damaging agents, thus providing at least one explanation for the poor prognosis associated with high GOLPH3 expression. Taken together, the data predict that inhibitors of the GOLPH3 pathway will have efficacy against cancer and will synergize with conventional therapeutics. Our mechanistic studies have allowed us to develop highly sensitive and specific assays for GOLPH3 pathway activity. Our preliminary data validate our primary assay for a high throughput screen to identify small molecule inhibitors of the pathway and further demonstrate our ability to validate, stratify, and determine the molecular targets of hit compounds. Here we propose a high throughput screen to identify inhibitors of the Golgi GOLPH3 pathway to serve as tools to study the role of the Golgi GOLPH3 pathway in cancer and as lead compounds for novel cancer therapeutics.

高通量筛选以鉴定高尔基体GOLPH 3通路的抑制剂 项目摘要 GOLPH 3是第一个从高尔基体运输到血浆的癌基因的例子 膜的GOLPH 3基因在人类癌症中经常扩增（例如56%的肺癌）， GOLPH 3的过表达驱动细胞培养物和癌症异种移植模型中的致癌转化。 此外，GOLPH 3的过表达在广泛的癌症谱中以高频率发生，并且在高水平的癌症中以高频率发生。 表达预示患者预后差。我们发现GOLPH 3是一种新的效应子， 磷脂酰肌醇PI 4P。我们发现GOLPH 3通过与PI 4P结合定位于高尔基体。GOLPH 3也 与一种非传统的肌球蛋白MYO 18 A结合，MYO 18 A将高尔基体与肌动蛋白细胞骨架连接起来。的 PI 4P/GOLPH 3/MYO 18 A/F-actin通路在高尔基体向质膜运输中的作用 拉伸力，以帮助囊泡从高尔基体出芽。这种张力也有助于使高尔基体 通过光学和电子显微镜观察到相似的外观。因此，高尔基体的形状是 贩运机制。GOLPH 3在模型系统中驱动致癌转化的能力是 依赖于PI 4P/GOLPH 3/MYO 18 A/F-actin途径。此外，我们已经表明，这种GOLPH 3 途径可以赋予对DNA损伤剂杀伤的抗性，从而提供了至少一种解释， GOLPH 3高表达与预后不良有关。总之，数据预测， GOLPH 3途径将具有抗癌功效，并将与常规治疗剂协同作用。我们 机制研究使我们能够开发出高灵敏度和特异性的GOLPH 3通路检测方法 活动我们的初步数据验证了我们用于高通量筛选以鉴定小分子的初步测定 抑制剂的途径，并进一步证明我们的能力，以验证，分层，并确定分子 击中化合物的目标。在这里，我们提出了一个高通量筛选，以确定抑制剂的高尔基GOLPH 3 作为研究高尔基体GOLPH 3通路在癌症中的作用的工具， 新的癌症治疗剂。