Integration of redox-balancing mechanisms in Pseudomonas aeruginosa biofilms

尽管已经开发了各种各样的抗微生物药物，但细菌和真菌感染仍然存在 流行于世界各地。微生物多细胞群落的形成，称为生物膜，是一种常见的 这些疾病的特征有助于他们的耐力。生物膜中的细胞由一个 细胞外基质，不同于液体培养的细胞，其代谢和 对抗菌药物的敏感性。我们的长期目标是定义以氧化还原为基础的综合监管 支持细菌在生物膜中生存的网络和代谢，重点是病原体 铜绿假单胞菌。我们假设发育中的生物膜中细胞氧化还原状态的变化会影响 多细胞行为、毒性和对抗生素的敏感性。一种标准化的菌落生物膜形态 化验有助于我们研究支持生物膜特定代谢和特征的机制 队形。我们将使用基本的分子方法与新开发和创新的技术相结合， 包括微米级生物膜的电化学和显微分析，以解决基本问题 关于使用氧化还原活性底物进行代谢和生物被膜的全球调节的问题 基质对氧化还原信号的反应：(1)铜绿假单胞菌是如何在氧气中平衡其细胞氧化还原状态的？ 生物膜的有限区域？(2)细胞信号的氧化还原控制如何决定整个群落 生物膜内的形态和细胞排列？(3)丙酮酸和乳酸的利用如何， 重要的能源和代谢中间体，有助于在生物膜中存活和代谢 宿主感染？由于铜绿假单胞菌是医院和囊性纤维化患者感染的主要原因， 我们的动机是这些机制作为治疗靶点的潜力。

Although a diverse arsenal of antimicrobial drugs has been developed, bacterial and fungal infections remain prevalent worldwide. The formation of multicellular communities of microbes, called biofilms, is a common feature of these diseases that contributes to their endurance. Cells in biofilms are held together by an extracellular matrix and differ from those grown in liquid cultures with respect to their metabolism and sensitivity to antimicrobial drugs. Our long-term goal is to define the integrated, redox-based regulatory networks and metabolisms that support survival of bacteria in biofilms, with a focus on the pathogen Pseudomonas aeruginosa. We hypothesize that the changing redox states of cells in developing biofilms affect multicellular behavior, virulence, and susceptibility to antibiotics. A standardized colony biofilm morphology assay facilitates our investigation of mechanisms that underpin biofilm-specific metabolism and feature formation. We will use basic molecular approaches combined with newly developed and innovative techniques, including electrochemical and microscopic analysis of biofilms at the micron scale, to address fundamental questions regarding the use of redox-active substrates for metabolism and the global regulation of biofilm matrix in response to redox cues: (1) How does P. aeruginosa balance its cellular redox state in the oxygen- limited regions of biofilms? (2) How does redox control of cellular signaling determine overall community morphology and cellular arrangement within the biofilm? (3) How does the use of pyruvate and lactate, important energy sources and metabolic intermediates, contribute to survival in biofilms and metabolism during host infection? As P. aeruginosa is a major cause of infections in hospitals and in patients with cystic fibrosis, we are motivated by the potential for these mechanisms to serve as therapeutic targets.