Synthetic biology for controlled release

摘要： 药物的简易生物合成与人类微生物组的操纵相结合的前景是 充满了治疗的可能性。然而，同样的警告存在于通过摄取传递药物 或注射适用于药物的微生物输送。特别是，给药的治疗方案必须 被确定为有效但无害的。多年来，确保长期交付的一种手段 特定数量的药物已经开发出颗粒、药丸或贴片，以维持受控或 将药物持续释放到系统中。我们提出了一种新的控制释放范式，其中 受控释放是由受控生物合成驱动的，而受控生物合成又依赖于潜在的模块化调节 机制。我们为治疗货物的表达建立了一个单独的“引擎”，依赖于高度 正交T7 RNA聚合酶(T7 RNAP)，并开发出各种导致调控基因的电路 表达在不同模式的治疗相关性，如恒定的动态平衡产生 药物浓度(目标1)。然后我们将这个“引擎”应用到氨基酸L-多巴的生产中 已知的益生菌菌株，E.coliNissle(目标2)。最后是益生菌的受控生产电路 在老鼠模型中引入物种是为了确定程序化的调节电路如何影响 药物在生物体中的药代动力学和药效学(目标3)。这些菌株最终要进行测试 在一种慢性进行性退行性帕金森氏病小鼠模型中，目前在我们的 爱荷华州立大学合作者实验室(AIM 3.3)。

Abstract: The prospects for the facile biosynthesis of drugs coupled with the manipulation of the human microbiome is fraught with therapeutic possibilities. However, the same caveats that exist for the delivery of drugs via ingestion or injection apply to the microbial delivery of drugs. In particular, a therapeutic regime for administration must be established that is efficacious but not harmful. For years, one means of ensuring the longer-term delivery of drugs in specified amounts has been to develop particles, pills, or patches that maintain the controlled or sustained release of drugs into the system. We propose a new paradigm for controlled release, in which controlled release is driven by controlled biosynthesis, which in turn relies on an underlying, modular regulatory mechanism. We establish a separate ‘engine’ for the expression of therapeutic cargoes, relying on the highly orthogonal T7 RNA polymerase (T7 RNAP), and develop a variety of circuits that lead to regulated gene expression in different patterns of therapeutic relevance, such as homeostatic production of constant concentrations of a drug (Aim 1). We then apply this ‘engine’ to the production of the amino acid L-DOPA in a known probiotic strain, E. coli Nissle (Aim 2). And then finally the controlled production circuitry in the probiotic species is introduced into mouse models in order to determine how programmed regulatory circuitry can impact the pharmacokinetics and pharmacodynamics of a drug in an organism (Aim 3). The strains are ultimately tested in a chronic progressive degenerative MitoPark mouse model of Parkinson’s disease currently being used in our collaborator’s laboratory at Iowa State University (Aim 3.3).