Optogenetic Control of Tumor Initiation and Tumor Progression in vivo

癌症研究和治疗发展严重依赖动物模型。中所需的关键功能之一 小鼠模型具有控制肿瘤发生和发展的能力。但是，现有方法不能控制 肿瘤将在何时何地形成和/或不允许调节特定的致癌信号模式 肿瘤进展。在这里，我们建议开发一种体内系统，它将提供对致癌因素的严格控制 对特定器官内的时间和位置进行精确控制的信号。实现时空联动 我们将使用一种新的光遗传策略来控制蛋白质的活性和基因的表达 光调节蛋白质。我们之前的研究由IMAT R21拨款资助，使我们能够开发出一种光- 调节(LightR)结构域，可以作为变构开关来控制蛋白质的活性。使用这种方法， 我们建议开发体内策略来调节致癌蛋白激酶和癌基因的表达。 它推动了肺癌的发生和发展，并介导了肺癌转移灶的发展 阿龙。其目标是建立一套工具，使研究人员能够模拟特定器官中的致癌信号 探讨其在肿瘤发生发展和肿瘤环境调控中的作用。作为模型系统 开发新技术，我们将开发一种工具包，使之能够调节致癌KRAS的表达 以及在小鼠肺中选择的位置精确计时的Src激酶活性。这些型号将允许 研究人员询问KRAS和Src在肺癌的发生、发展和扩散中的作用 其他地点。我们还将开发一个系统，使地方监管的致癌信号促进 循环癌细胞在肺内的转移。该方法将允许对特定阶段的 转移过程。这将为询问促进转移的致癌信号提供新的工具。 癌细胞的能力和体内转移前肿瘤生态位的调节。建议的设计 系统将确保它们适用于不同的癌症模型。

Cancer research and therapy development rely heavily on animal models. One of the key features desired in a mouse model is ability to control tumor initiation and progression. However, existing methods cannot control where and when tumor will form and/or do not allow regulation of specific oncogenic signaling patterns driving tumor progression. Here we propose to develop an in vivo system that will provide tight control of oncogenic signaling with precise control of timing and location within a specific organ. To achieve spatial and temporal control of protein activity and gene expression we will use a novel optogenetic strategy that employs engineered light-regulated proteins. Our previous studies funded by an IMAT R21 grant allowed us to develop a Light- Regulated (LightR) domain that can function as allosteric switch to control protein activity. Using this method, we propose to develop in vivo strategy for regulation of oncogenic protein kinases and expression of oncogenes that drive initiation and progression of lung cancer as well as mediate development of metastatic lesions in the lung. The goal is to build a set of tools that will allow researcher to model oncogenic signaling in a specific organ and interrogate its role in tumor development and regulation of tumor environment. As model systems for development of new technology, we will develop a toolkit that enables regulation of oncogenic KRas expression and Src kinase activity at a selected location in mouse lungs with precise timing. These models will allow researchers to interrogate the role of KRas and Src in initiation of lung cancer, its progression, and spreading to other locations. We will also develop a system that will enable local regulation of oncogenic signaling promoting metastasis of circulating cancer cells in the lung. This method will enable regulation of specific stages of metastatic process. It will provide new tools for interrogation of oncogenic signaling in promoting metastatic ability of cancer cells and regulation of the pre-metastatic tumor niche in vivo. The design of the proposed systems will ensure their application for different cancer models.