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.

项目概要 病原微生物群落内细胞之间的化学相互作用对 表型状态；许多化学信号、毒力因子和氧化还原活性物质都会影响 细菌成分的毒力潜力和免疫细胞的反应。详细了解如何 “微观地理”通过社会线索、化学战和其他系统的传播影响细胞行为 因此，诸如氧气和资源限制之类的属性将提供潜在有价值的新长期 对抗感染的关键策略。能够精确定位小的细菌聚集体 感兴趣的微生物群落内定义的物理和表型属性将使研究成为可能 细胞之间随时间变化的空间相互作用，例如在细胞的传播和进展中可能发生的相互作用 感染，以及已建立的微生物组内的动态重组。 在这里，我们建议开发一个用于研究社会微生物学和其他领域的技术平台 病原微生物群落中基于化学的属性，其中细菌菌落，以及某些 例如，来自具有确定种群规模/密度、形状、排列和 社会微生物学状态将被组织在基于多孔蛋白质的微容器中 3D 打印 可操纵的光纤尖端。该光极平台将提供报告和 通过微容器进行轻松的化学交换，可控地扰动感兴趣的微生物环境 墙壁，并将通过检测荧光来获取微观生态系统的详细分子信息 转录报告基因和其他细胞探针，以及一氧化氮和一氧化氮的工程化学报告基因 活性氧结合在微容器壁中。作为平台的验证，我们将评估 群体感应和群体依赖性抗生素耐药性传播的时空属性 纤维安装的 3D 打印 Pa 聚集体和模型微生物群落之间的双向 作为视觉上可访问的文化中聚合体的物理固定集合体进行维护 倒置显微镜系统。作为将方法扩展到细菌递送的概念验证 社区到体内环境，我们将在大格式成像上 3D 制造定义的 Pa 群体 光纤束。将评估纤维安装群体和固定群体之间的表型相互作用 Pa/Sa 文化。如果成功，从这些高风险、高收益研究中获得的能力将具有 各种体外模型和体内微生物组的中长期应用，包括慢性 伤口、胃肠道和呼吸系统感染。