A Cancer Rainbow Mouse for the Simultaneous Assessment of Multiple Oncogenes

描述（由申请人提供）：了解肿瘤发生的遗传机制将为开发新一代更好、更有效的癌症治疗方法提供基础。为了解决这一根本问题，需要比现有技术更上级的新技术。现有技术的小鼠模型能够以细胞类型特异性方式测试癌基因的随机激活，并提供用于测试靶向疗法的机制。尽管有这些可取的特点，其有限的可及性和可行性排除了他们在大多数实验室的实施。此外，随着致癌基因的不断增加，需要可靠和有效的技术，可以同时评估几个致癌基因，以促进快速分析致瘤性及其对治疗的反应。该应用通过开发创新的转基因小鼠平台（Crabow）来快速、高效且具有成本效益地创建肿瘤发生的小鼠模型，从而为所有这些问题提供了解决方案。Crabow技术将使多个用户可选择的癌基因被整合到单个转基因小鼠中，从而每个癌基因将基于独特的表位标签和单独表达的光谱可分辨荧光蛋白被识别。使用Cre诱导的随机激活策略，Crabow技术能够在同一组织中以单细胞分辨率同时评估每个癌基因的贡献。此外，克隆来源的肿瘤 细胞群可以通过繁殖到大量经验证的细胞类型特异性Cre小鼠系来研究任何组织和细胞类型。该技术的灵活性的基础是一种新的质粒构建体，用于掺入感兴趣的癌基因。使用大多数生物学实验室中可用的分子生物学技能，可以容易地修饰彩虹质粒用于癌基因靶向。然后，可以在现场快速生产彩虹转基因小鼠，或者从其他研究者或储存库获得。通过以下具体目标证明了该Crabow平台技术的实用性和多功能性：（1）建立用于癌症rainbow转基因设计和构建的灵活且快速的策略，（2）生成并验证癌症rainbow转基因小鼠，以及（3）使用癌症rainbow转基因小鼠生成组织特异性肿瘤。癌症彩虹技术通过提供一种同时监测多个基因活性的综合方法，以及一种快速筛选新型靶向癌症疗法的手段，解决了目前技术在单个小鼠中的缺点。因此，癌症彩虹技术将为研究肿瘤发生提供一个范式转变。 公共卫生相关性：这项应用将建立一个强大的，但易于使用的技术，将取代以前的嵌合肿瘤小鼠模型。因此，靶向这些癌基因的癌症治疗策略可以很容易且经济地进行测试，从而产生更有效的患者治疗和更好的临床结果。

DESCRIPTION (provided by applicant): Understanding the genetic mechanisms which underlie tumor development will provide a foundation for developing new generations of better and more effective cancer therapies. To address this fundamental issue, new technologies are needed that are superior to those currently available. Current state of the art mouse models enable testing the stochastic activation of oncogenes in a cell type specific fashion and provide a mechanism for testing targeted therapies. Despite these desirable features, their limited accessibility and feasibility precludes their implementation in most laboratories. Moreover, as the list of oncogenes continues to grow, reliable and efficient technologies are needed that can assess several oncogenes simultaneously in order to facilitate rapid analysis of tumorigenicity and their responses to therapy. This application provides a solution to all these issues by developing an innovative transgenic mouse platform (Crainbow) to rapidly, efficiently, and cost effectively create mouse models of tumorigenesis. The Crainbow technology will enable multiple user selectable oncogenes to be incorporated into a single transgenic mouse whereby each oncogene will be identifiable on the basis of a unique epitope tag and a separately expressed spectrally resolvable fluorescent protein. Using a strategy of Cre induced stochastic activation, the Crainbow technology enables the contribution of each oncogene to be assessed coincidentally in the same tissue with single cell resolution. Additionally, clonally derived tumor cell populations can be studied in any tissue and cell type by breeding to the plethora of validated cell type specific Cre mouse lines. Underlying the flexibility of this technology is a novel plasmid construct for incorporating the oncogenes of interest. The Crainbow plasmid can be easily modified for oncogene targeting using the molecular biology skills available in most all biology laboratories. Crainbow transgene mice can then be produced rapidly on site or obtained from other investigators or repositories. The utility and versatility of this Crainbow platform technology is demonstrated through the following specific aims which: (1) Establish a flexible and rapid strategy for cancer rainbow transgene design and construction, (2) Generate and validate cancer rainbow transgenic mice, & (3) Generate tissue specific tumors using cancer rainbow transgenic mice. The Cancer rainbow technology addresses in a single mouse shortcomings of current technology by providing a comprehensive method to simultaneously monitor the activity of multiple genes, and a means to also rapidly screen novel targeted cancer therapies. Consequently, the Cancer rainbow technology will provide a paradigm shift for studying tumorigenesis. PUBLIC HEALTH RELEVANCE: This application will establish a powerful but easily employed technology that will supplant previous chimeric tumor mouse models. Therefore, cancer treatment strategies targeting these oncogenes can be easily and economically tested, resulting in more effective patient therapies and better clinical outcomes.