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).

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，对于通过摄入递送药物存在的相同注意事项 或注射剂应用于微生物药物递送。特别地，用于施用的治疗方案必须 这是有效的，但不是有害的。多年来，确保长期提供 药物在特定数量一直是开发颗粒，药丸，或补丁，保持控制或 药物持续释放到系统中。我们提出了一种新的控制释放模式，其中 控释由受控的生物合成驱动，而生物合成又依赖于潜在的、模块化的调控机制。 机制我们建立了一个单独的'引擎'的表达治疗货物，依靠高度 正交T7 RNA聚合酶（T7 RNAP），并开发出各种电路，导致调控基因 不同模式的治疗相关性表达，如恒定的 药物浓度（目标1）。然后，我们将这种“引擎”应用于氨基酸L-DOPA的生产， 已知益生菌菌株，E. coli Nissle（Aim 2）。最后是益生菌中的受控生产回路 物种引入小鼠模型，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）。这些菌株最终被测试 在我们目前使用的帕金森病慢性进行性退化MitoPark小鼠模型中， 爱荷华州州立大学的合作者实验室（目标3.3）。