Optogenetic Control of Tumor Initiation and Tumor Progression in vivo

癌症研究和治疗开发严重依赖于动物模型。在一个关键的功能，希望在一个 小鼠模型是控制肿瘤发生和发展的能力。然而，现有方法无法控制 肿瘤将在何时何地形成和/或不允许调节特定致癌信号传导模式， 肿瘤进展。在这里，我们建议开发一个体内系统，将提供严格的控制致癌 在特定器官内精确控制时间和位置的信号传导。为了实现空间和时间 为了控制蛋白质活性和基因表达，我们将使用一种新的光遗传学策略， 光调节蛋白我们以前的研究由IMAT R21赠款资助，使我们能够开发一种光- 调节（LightR）结构域，可作为变构开关控制蛋白质活性。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 这些基因驱动肺癌的发生和发展，并介导转移性病变的发展。 肺。目标是建立一套工具，使研究人员能够在特定器官中模拟致癌信号 并探讨其在肿瘤发生和肿瘤环境调节中的作用。作为模型系统， 随着新技术的发展，我们将开发一个工具包，使致癌KRas表达的调控 和Src激酶活性在小鼠肺中的选定位置以精确的定时。这些模型将允许 研究人员询问KRas和Src在肺癌发生、进展和扩散中的作用。 其他地点。我们还将开发一个系统，使致癌信号的局部调节，促进 循环癌细胞在肺中的转移。这种方法将能够调节特定阶段的 转移过程它将为询问致癌信号在促进转移中的作用提供新的工具。 癌细胞的能力和体内转移前肿瘤小生境的调节。建议的设计 系统将确保其应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.