Deployable 3D-printed Cellular Communities for Characterizing Bacterial Social Cues and Chemical Warfare

可部署的 3D 打印细胞群落，用于表征细菌社交线索和化学战

• 批准号: 10005373
• 负责人: Eric V. Anslyn
• 金额: $ 18.53万
• 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: 2019-09-01 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10005373
• 关键词: 3-Dimensional; 3D Print; Antibiotic Resistance; Bacteria; Behavior; Cell Line; Cells; Chemical Engineering; Chemical Warfare; Chemicals; Cleaved cell; Communities; Complex; Cues; Detection; Development; Disease; Ecosystem; Environment; Face; Fiber; Fiber Optics; Fluorescence; Gastrointestinal tract structure; Genetic Transcription; Geography; Goals; Grant; Heterogeneity; Image; Immobilization; Immune response; In Vitro; Infection; Mammalian Cell; Methodology; Methods; Microscope; Microscopic; Modeling; Molecular; Nitric Oxide; Optics; Oxidation-Reduction; Oxygen; Pathogenicity; Phenotype; Play; Population; Population Sizes; Positioning Attribute; Proteins; Pseudomonas aeruginosa; Reactive Oxygen Species; Reporter; Reporting; Resolution; Resources; Respiratory System; Role; Shapes; Signal Transduction; Staphylococcus aureus; Structure; System; Technology; Time; Time Study; Validation; Variant; Virulence; Virulence Factors; Work; bacterial community; base; cell behavior; cell community; chronic infection; combat; comparative; density; fighting; high risk; in vitro Model; in vivo; instrumentation; interest; macrophage; microbial; microbial community; microbiome; optical fiber; pathogenic bacteria; quorum sensing; response; social; spatiotemporal; trait; transmission process; wound

Project Summary The chemical interplay between cells within pathogenic microbial communities has profound effects on phenotypic states; numerous chemical signals, virulence factors, and redox-active species influence both the virulence potential of bacterial constituents and the response of immune cells. Detailed understanding how “micro-geography” impacts cellular behavior via transmission of social cues, chemical warfare, and other system attributes such as oxygen and resource limitation would therefore offer potentially valuable new long-term strategies key for combatting infection. A capability to precisely position small, bacterial aggregates having defined physical and phenotypic attributes within microbial communities of interest would enable the study time-variant spatial interactions between cells, such as those that may occur in the spread and progression of infections, as well as dynamic reorganization within established microbiomes. Here, we propose development of a technology platform for investigating sociomicrobiology and other chemically based attributes in pathogenic microbial communities in which bacterial colonies and, in some instances, cells from mammalian lines of defined population sizes/densities, shapes, arrangements, and sociomicrobiological status will be organized in porous protein-based microcontainers 3D printed on maneuverable optical fiber tips. This optrode platform will provide capabilities both for reporting on and controllably perturbing, microbial environments of interest by facile chemical exchange through microcontainer walls and will acquire detailed molecular information on microscopic ecosystems via detection of fluorescent transcriptional reporters and other cellular probes, as well as engineered chemical reporters for nitric oxide and reactive oxygen species incorporated in microcontainer walls. As validation of the platform, we will evaluate spatiotemporal attributes for transmission of quorum sensing and population-dependent antibiotic resistance bi-directionally between fiber-mounted 3D-printed Pa aggregates and model microbiome communities maintained as a physically stationary ensemble of aggregates in visually accessible cultures maintained on an inverted microscope system. As proof-of-concept for extending methodologies to delivery of bacterial communities to in vivo environments, we will 3D fabricate defined populations of Pa on large-format imaging fiber-optic bundles. Phenotypic interplay will be evaluated between fiber-mounted populations and stationary cultures of Pa/Sa. If successful, capabilities that derive from these high-risk, high-benefit studies will have medium- and long-term applications to various in vitro models and in vivo microbomes, including chronic infections in wounds, gastrointestinal tract, and respiratory system.

项目摘要 致病微生物群落中细胞之间的化学相互作用对 表型状态；许多化学信号，病毒因子和氧化还原活性物种都会影响 细菌的毒力潜力构成免疫细胞的反应。详细了解如何 “微观地理”通过传播社会提示，化学战和其他系统影响细胞行为 因此，氧气和资源限制等属性将提供潜在的有价值的新型长期 策略的关键是打击感染。精确定位小的能力，具有细菌质量 感兴趣的微生物群落中定义的物理和表型属性将使研究能够 细胞之间的时间变化空间相互作用，例如可能发生在传播和进展中的空间相互作用 在已建立的微生物组中的感染以及动态重组。 在这里，我们建议开发一个技术平台，以调查社会生物学和其他 致病微生物群落中基于化学的属性，其中细菌菌落和某些菌落中的属性 实例，来自定义的人口大小/密度，形状，布置和的细胞 社会生物学状态将在基于多孔蛋白质的微观核心剂3D上组织 可操纵的光纤技巧。此Optrode平台将提供有关报告和报告的功能 通过微牙本质可控化学交换，可控性，感兴趣的微生物环境 墙壁并将通过检测荧光获取微观生态系统的详细分子信息 转录记者和其他蜂窝探针，以及一氧化氮和一氧化氮的工程化学记者 掺入微端壁中的活性氧。作为平台的验证，我们将评估 时空属性，用于传播群体传感和种群依赖性抗生素耐药性 在纤维上安装的3D打印PA聚集体和模型微生物组群落之间的双向方向 在维持在视觉上可接近的文化中的骨料的物理固定合奏保持在一个维持在 倒立显微镜系统。作为扩展细菌递送方法的概念验证 社区到体内环境，我们将在大幅面成像上3D织物定义的PA的群体 光纤束。表型相互作用将在纤维安装的种群和固定型之间进行评估 PA/SA的文化。如果成功，则从这些高风险，高效力研究中获得的能力将具有 在各种体外模型和体内微生物中的中期和长期应用，包括慢性 伤口，胃肠道和呼吸系统的感染。