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.

用于鉴定高尔基体 GOLPH3 通路抑制剂的高通量筛选 项目概要 GOLPH3 是第一个在从高尔基体到血浆的运输中起作用的癌基因的例子 膜。 GOLPH3 基因在人类癌症（例如 56% 的肺癌）中频繁扩增，并且 GOLPH3 的过度表达可驱动细胞培养和癌症异种移植模型中的致癌转化。 此外，GOLPH3 的过度表达在多种癌症中以高频率发生，且高表达率 表达预示患者预后不良。我们发现 GOLPH3 作为一种新的效应子 磷酸肌醇PI4P。我们发现 GOLPH3 通过与 PI4P 结合定位于高尔基体。 GOLPH3也 与非常规肌球蛋白 MYO18A 结合，将高尔基体与肌动蛋白细胞骨架连接起来。这 PI4P/GOLPH3/MYO18A/F-肌动蛋白通路通过应用高尔基体到质膜的运输发挥作用 张力以协助囊泡从高尔基体出芽。这种拉力也有助于赋予高尔基体 通过光学和电子显微镜观察，具有熟悉的外观。因此，高尔基体的形状是 贩卖机制。 GOLPH3 在模型系统中驱动致癌转化的能力是 依赖于 PI4P/GOLPH3/MYO18A/F-肌动蛋白通路。此外，我们还证明了 GOLPH3 途径可以赋予对 DNA 损伤剂杀伤的抵抗力，从而提供至少一种解释 GOLPH3 高表达相关的不良预后。综上所述，数据预测抑制剂 GOLPH3 通路将具有抗癌功效，并将与传统疗法产生协同作用。我们的 机制研究使我们能够开发出针对 GOLPH3 通路的高度敏感和特异的检测方法 活动。我们的初步数据验证了我们用于高通量筛选的主要检测方法，以识别小分子 该途径的抑制剂，并进一步证明我们验证、分层和确定分子水平的能力 命中化合物的目标。在这里，我们提出了一种高通量筛选来鉴定高尔基体 GOLPH3 的抑制剂 途径作为研究高尔基体 GOLPH3 途径在癌症中的作用的工具，并作为先导化合物 新的癌症疗法。