Split Proteins As Boolean Circuits and Drugs of a New Kind

描述（由申请人提供）：在尤里卡的申请中，标题为“作为布尔电路的分裂蛋白质和新型药物”，我们提出验证、优化和实施一种治疗“复杂”疾病的新方法。在这种称为缺失特异性靶向（DST）的方法中，异常细胞以一种新的方式通过其DNA中的突变变化，特别是纯合DNA缺失（HD）来靶向。例如，许多癌症已被证明具有HD。（虽然癌症是一个相关的例子，但相同的“DST”逻辑适用于任何可以显示含有HD的不期望的细胞谱系。换句话说，DST策略也与癌症以外的疾病有关。纯合缺失的一个显著特性是这种“阴性”靶标不能恢复，从而作为治疗的不可变的路标。要瞄准HD，即，DST策略以一种新颖的方式将现有的和新的方法结合在一起，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性大范围核酸酶。DST策略还采用反馈机制，该反馈机制从作为布尔或门操作的电路接收输入，并且涉及分裂大范围核酸酶的激活，其破坏正常（非靶）细胞中的DST载体。DST的逻辑使治疗选择性的基本上无限增加成为可能。我们建议在酵母S中测试和开发DST。酿酒酵母，一种遗传上易于处理的生物，其中复杂的操作更快， 比哺乳动物细胞更容易。该项目的这一阶段之后将进行测试， 在培养的哺乳动物细胞中，以及此后在具有特定疾病的完整小鼠中的（优化的）电路。 项目叙述：尤里卡申请中提出的研究旨在验证、开发和实施一种治疗“复杂”疾病的新方法。这种称为缺失特异性靶向（DST）的方法使得靶向异常细胞中的纯合缺失（HD）成为可能，例如，在癌细胞（但不仅仅是癌细胞）中。DST策略在临床环境中的最终实施可能被证明对一系列HD相关疾病具有治愈性，此外基本上没有副作用。

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.