Split Proteins As Boolean Circuits and Drugs of a New Kind

描述(由申请人提供)：在这份名为“分离蛋白质作为布尔电路和新型药物”的尤里卡申请中，我们建议验证、优化和实施一种治疗“复杂”疾病的新方法。在这种被称为缺失特异性靶向(DST)的方法中，通过DNA突变，特别是纯合子DNA缺失(HDs)，以一种新颖的方式将异常细胞靶向。例如，许多癌症已被证明含有HDs。(虽然癌症是一个相关的例子，但同样的“DST”逻辑也适用于任何可以被证明含有HDs的不良细胞系。换句话说，DST策略也与癌症以外的疾病相关。)纯合缺失的一个显著特征是，这个“阴性”靶点不能逆转，从而成为治疗的一个不变的路标。为了针对HD，即缺乏的“靶”，DST策略以一种新颖的方式结合了现有和新的方法，包括泛素融合技术、分裂泛素分析、锌指DNA识别蛋白和分裂限制性巨核酸酶。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，并涉及分裂巨核酸酶的激活，该分裂巨核酸酶破坏正常(非靶标)细胞中的DST载体。DST的逻辑使得基本上无限增加治疗的选择性成为可能。我们建议首先在酿酒酵母中测试和开发dst，这是一种遗传上易于处理的生物，在这种生物中，复杂的操作速度更快。 而且比在哺乳动物细胞中更容易。在项目的这一阶段之后，将测试所产生的 (优化)在哺乳动物细胞培养中的电路，以及此后在患有特定疾病的完整小鼠中的电路。 项目简介：在这项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。这种方法被称为缺失特异性靶向(DST)，使得在异常细胞中靶向纯合缺失(HDs)成为可能，例如，在癌症(但不仅仅是癌症)细胞中。最终在临床环境中实施DST策略可能会被证明对一系列与HD相关的疾病是治愈的，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.