Split Proteins As Boolean Circuits and Drugs of a New Kind

将蛋白质拆分为布尔电路和新型药物

• 批准号: 7661504
• 负责人: ALEXANDER J VARSHAVSKY
• 金额: $ 32.1万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7661504
• 关键词: Abnormal Cell; Adverse effects; Biological Assay; Cell Lineage; Cells; Clinical; Complex; DNA; Disease; Feedback; Logic; Malignant Neoplasms; Mammalian Cell; Methodology; Mus; Organism; Pharmaceutical Preparations; Property; Proteins; Saccharomyces cerevisiae; Staging; Techniques; Test Result; Testing; Ubiquitin; Yeasts; Zinc Fingers; cancer cell; design; novel; novel strategies; vector

DESCRIPTION (provided by applicant): IIn this EUREKA application, entitled "Split Proteins As Boolean Circuits and Drugs of a New Kind", we propose to verify, optimize, and implement a new approach to therapy of "complex" diseases. In this approach, termed deletion-specific targeting (DST), the abnormal cells are targeted, in a novel way, through mutational changes in their DNA, specifically homozygous DNA deletions (HDs). For example, many cancers have been shown to harbor HDs. (Although cancer is a relevant example, the same "DST" logic applies to any undesirable cell lineage that can be shown to contain HDs. In other words, the DST strategy is also relevant to diseases other than cancer.) A salient property of a homozygous deletion is that this "negative" target cannot revert, thereby serving as an immutable signpost for therapy. To target an HD, i.e., a "target" that is absent, the DST strategy brings together, in a novel way, both existing and new methodologies, including the ubiquitin fusion technique, split-ubiquitin assay, zinc-finger DNA-recognizing proteins and split restriction meganucleases. The DST strategy also employs a feedback mechanism that receives input from a circuit operating as a Boolean OR gate and involves the activation of split meganucleases, which destroy DST vector in normal (non-target) cells. The logic of DST makes possible an essentially unlimited increase in the selectivity of therapy. We propose to test and develop DST initially in the yeast S. cerevisiae, a genetically tractable organism in which sophisticated manipulations are faster and easier than in mammalian cells. This stage of the project will be followed by tests of resulting (optimized) circuits in mammalian cells in culture, and thereafter in intact mice with specific diseases. Project Narrative: The studies proposed in this EUREKA application are designed to verify, develop and implement a new approach to therapy of "complex" diseases. This approach, termed deletion-specific targeting (DST), makes possible the targeting of homozygous deletions (HDs) in abnormal cells, e.g., in cancer (but not only cancer) cells. The eventual implementation of DST strategy in a clinical setting may prove to be curative with a range of HD-relevant diseases, and in addition substantially free of side effects.

描述（由申请人提供）：iin这个Eureka应用程序，标题为“将蛋白分开为新型的布尔电路和药物”，我们建议验证，优化和实施一种新的治疗“复杂”疾病的方法。在这种方法中，称为缺失特异性靶向（DST），异常细胞通过其DNA的突变变化（特别是纯合DNA缺失（HDS））以新颖的方式靶向。例如，许多癌症已被证明具有HDS。 （尽管癌症是一个相关的例子，但相同的“ DST”逻辑适用于任何可能证明包含HDS的不良细胞谱系。换句话说，DST策略也与癌症以外的其他疾病有关。）纯合删除是，这个“负”靶标无法恢复，从而成为治疗的不可变路。为了靶向HD，即缺乏的“目标”，DST策略以一种新颖的方式将现有方法和新方法汇集在一起​​，包括泛素融合技术，分裂泛素测定，锌指DNA识别蛋白和分裂限制巨核。 DST策略还采用了一种反馈机制，该机制从作为布尔或门的电路中接收输入，并涉及分裂的巨核酶的激活，该电路在正常（非目标）细胞中破坏了DST载体。 DST的逻辑使治疗选择性基本上无限。我们建议最初在酵母菌S.酿酒酵母中测试和开发DST，这是一种遗传上的有机体，其中复杂的操纵更快 比在哺乳动物细胞中更容易。该项目的这一阶段将进行结果测试 （优化的）培养哺乳动物细胞中的电路，此后在具有特定疾病的完整小鼠中。 项目叙述：本eureka应用中提出的研究旨在验证，开发和实施一种新的“复杂”疾病治疗方法。这种方法称为缺失特异性靶向（DST），使得异常细胞中的纯合缺失（HDS）的靶向可能，例如在癌症（而是癌症）细胞中。在临床环境中，最终实施DST策略可能被证明是通过一系列与HD相关的疾病治愈的，此外还没有副作用。