High Throughput Screen to Identify Inhibitors of the Golgi GOLPH3 Pathway

高通量筛选用于鉴定高尔基体GOLPH3途径的抑制物 项目总结 GOLPH3是第一个在高尔基体向血浆转运过程中起作用的癌基因 薄膜。GOLPH3基因在人类癌症(例如56%的肺癌)和 GOLPH3的过表达推动了细胞培养和异种移植癌症模型的致癌转化。 此外，GOLPH3的高表达在广泛的癌症和高 表达预示着患者预后较差。我们发现GOLPH3是一种新型的 磷脂酰肌醇PI4P。我们发现GOLPH3通过与PI4P结合定位于高尔基体。GOLPH3还 结合一种非传统的肌球蛋白MYO18A，它将高尔基体与肌动蛋白细胞骨架联系起来。这个 PI4P/GOLPH3/MYO18A/F-肌动蛋白途径在高尔基体到质膜转运中的作用 帮助来自高尔基体的囊泡萌发的拉力。这种张力也有助于为高尔基人提供 光镜和电子显微镜下可见熟悉的外观。因此，高尔基的形状是 贩运机制。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，我们已经证明了这个GOLPH3 通路可以抵抗DNA损伤剂的杀戮，因此至少提供了一种解释 预后不良与GOLPH3高表达有关。综上所述，这些数据预测， GOLPH3途径将对癌症有效，并将与传统疗法协同作用。我们的 机制研究使我们能够开发出对GOLPH3途径高度敏感和特异的分析方法 活动。我们的初步数据验证了我们用于高通量筛选以识别小分子的初步试验 途径的抑制物，并进一步展示我们验证、分层和确定分子的能力 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 作为工具研究高尔基体GOLPH3途径在癌症中的作用以及作为先导化合物 新的癌症疗法。

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.