Synthetic biology for controlled release

控制释放的合成生物学

• 批准号: 10376300
• 负责人: Andrew D Ellington
• 金额: $ 34.53万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-05-04 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10376300
• 关键词: Amino Acids; Anabolism; Automobile Driving; Bacteria; Base Pairing; Biological Assay; Blood; Chronic; Coupled; Data; Development; Disease; Drug Delivery Systems; Drug Kinetics; Elements; Engineered Probiotics; Ensure; Enzymes; Escherichia coli; Feedback; Gene Expression; Health; Human; Human Microbiome; Implant; In Vitro; Ingestion; Injections; Intervention; Iowa; Knowledge; Laboratories; Lead; Levodopa; Mathematics; Mediating; Messenger RNA; Methods; Mixed Function Oxygenases; Modeling; Mus; Organism; Output; Parkinson Disease; Pattern; Pharmaceutical Preparations; Physiologic pulse; Positioning Attribute; Probiotics; Production; Proteins; RNA; Regimen; Solid; Specific qualifier value; System; T7 RNA polymerase; Testing; Therapeutic; Time; Transgenic Mice; Universities; base; controlled release; design-build-test; experimental study; gut colonization; metabolic engineering; microbial; microbiome; microorganism; mitopark mouse; mouse model; novel; particle; pharmacokinetics and pharmacodynamics; pill; pre-clinical; synthetic biology; transcription factor

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).

抽象的： 药物的容易生物合成的前景以及人类微生物组的操纵 充满治疗可能性。但是，通过摄入提供药物的同样的警告 或注射适用于药物的微生物输送。特别是，必须进行治疗制度 确定这是有效但无害的。多年来，一种确保长期交付的方法 指定量的药物是开发维持受控或 持续将药物释放到系统中。我们提出了一个新的范式，用于受控释放，其中 受控释放是由受控的生物合成驱动的，这反过来依赖于下面的模块化调节 机制。我们建立了一个单独的“引擎”来表达治疗性货物，依靠高度 正交T7 RNA聚合酶（T7 RNAP），并发展出多种电路，导致调节基因 以不同的治疗相关方式表达表达，例如常数的体内平衡产生 药物的浓度（目标1）。然后，我们将此“引擎”应用于氨基酸L-DOPA的生产 已知的益生菌菌株，大肠杆菌Nissle（AIM 2）。然后最后是益生菌中受控的生产电路 将物种引入鼠标模型，以确定编程的调节电路如何影响 在有机体中药物的药代动力学和药效学（AIM 3）。菌株最终经过测试 目前在我们 爱荷华州立大学合作者的实验室（AIM 3.3）。