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

项目摘要 病原微生物群落内细胞之间的化学相互作用对微生物的生长有着深远的影响。 表型状态;许多化学信号，毒力因子和氧化还原活性物质影响这两个 细菌成分的毒力潜力和免疫细胞的反应。详细了解如何 “微观地理学”通过社会线索、化学战和其他系统的传递来影响细胞行为。 因此，氧气和资源限制等属性将提供潜在的有价值的新的长期 对抗感染的关键策略。精确定位小的细菌聚集体的能力， 在感兴趣的微生物群落中确定的物理和表型属性将使研究 细胞之间的时变空间相互作用，例如那些可能发生在细胞扩散和进展中的相互作用。 感染，以及建立微生物组内的动态重组。 在这里，我们建议开发一个技术平台，用于研究社会微生物学和其他 在致病微生物群落中，细菌菌落，在某些情况下， 在某些情况下，来自具有确定的群体大小/密度、形状、排列和分布的哺乳动物系的细胞， 社会微生物学状态将被组织在3D打印的多孔蛋白质微容器中， 可拆卸的光纤尖端。该光电管平台将提供报告和 通过微容器的简易化学交换可控地扰动感兴趣的微生物环境 并将通过检测荧光物质来获取微观生态系统的详细分子信息。 转录报告基因和其它细胞探针，以及用于一氧化氮的工程化学报告基因， 反应性氧物质结合在微容器壁中。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 作为聚集体的物理上固定的集合体保持在视觉上可访问的培养物中， 倒置显微镜系统作为将方法扩展到细菌递送的概念验证， 社区在体内环境中，我们将3D制造大格式成像上的Pa定义种群 光纤束将评价纤维固定群体和固定群体之间的表型相互作用。 文化的Pa/Sa。如果成功，从这些高风险、高收益研究中获得的能力将具有 中期和长期应用于各种体外模型和体内微生物，包括慢性 伤口、胃肠道和呼吸系统感染。

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.