Synthetic biology for controlled release

控制释放的合成生物学

• 批准号: 9926117
• 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 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9926117
• 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; 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： 药物生物合成与人类微生物组操纵相结合的前景是 充满了治疗的可能性。然而，通过摄入输送药物也存在同样的警告 或注射剂适用于微生物输送药物。特别是，给药治疗方案必须 确定有效但无害。多年来，确保长期交付的一种手段 特定数量的药物已开发出颗粒、药丸或贴剂，以维持受控或 药物持续释放到系统中。我们提出了一种新的受控释放范式，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监管 机制。我们建立了一个单独的“引擎”来表达治疗货物，依赖于高度 正交T7 RNA聚合酶（T7 RNAP），并开发多种导致调控基因的电路 以不同的治疗相关性模式表达，例如恒定的稳态产生 药物浓度（目标 1）。然后，我们将这个“引擎”应用于氨基酸 L-DOPA 的生产 已知益生菌菌株，大肠杆菌 Nissle（目标 2）。最后是益生菌中的受控生产电路 将物种引入小鼠模型中，以确定编程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）。最终对菌株进行测试 在慢性进行性退行性帕金森病 MitoPark 小鼠模型中，目前我们正在使用 爱荷华州立大学合作者实验室（目标 3.3）。