Exploring mechanical mechanisms of antibiotic resistance

探索抗生素耐药性的机械机制

• 批准号: 10276887
• 负责人: Enrique Rojas
• 金额: $ 32.26万
• 依托单位: NEW YORK UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10276887
• 关键词: Address; Antibiotic Resistance; Antibiotics; Bacteria; Bacterial Infections; Biochemical; Biological; Biological Assay; Biophysics; Cell Death; Cell Wall; Cells; Dependence; Explosion; Fracture; Gram-Positive Bacteria; Growth; Hydrostatic Pressure; Knowledge; Measures; Mechanics; Membrane; Microbiology; Microfluidics; Microscopy; Modeling; Molecular; Monitor; Pathogenesis; Physiological Processes; Physiology; Process; Property; Research; Resistance; Structure; Techniques; Teichoic Acids; Weight-Bearing state; bacterial resistance; base; cell envelope; combat; exoskeleton; innovation; mechanical force; mechanical properties; novel; pathogenic bacteria; protein protein interaction; resistance mechanism; theories; tool

Summary Traditionally, studies of antibiotic resistance have focused on evolutionary and molecular mechanisms of resistance. However, many of our best antibiotics target the bacterial cell envelope, which is a mechanically robust, structural exoskeleton for the cell. Ultimately, these antibiotics cause cell death by weakening the envelope enough to cause explosion of the cell by the large, hydrostatic pressure within it. Despite the central mechanical importance of the cell envelope, we have little understanding of which molecules and moieties within it are critical for its load-bearing capacity. Addressing this question would transform our understanding of antibiotic resistance. A primary reason for this gap in our knowledge is the formidable challenge of applying mechanical forces to single bacterial cells while monitoring their physiology. The proposed research will address this obstacle by applying innovative, highly precise, high-throughput microfluidics and microscopy-based assays to measure the mechanical properties of two of the major cell envelope components in bacteria, the outer membrane and the cell wall. These assays will be combined with molecular and cell biological techniques, and biophysical theory, to explore an emerging paradigm within microbiology: that bacteria control antibiotic resistance by adaptively tuning the mechanical properties of their cell envelope. First, building on the recent landmark finding that the outer membrane confers antibiotic resistance to bacteria because of its mechanical strength, the dependence of outer membrane stiffness and mechanical antibiotic resistance on the fine-scale biochemical composition of the outer membrane will be systematically measured. Next, the mechanism of outer membrane vesiculation (a process underlying antibiotic resistance and pathogenesis) will be investigated by combining a theoretical mechanical model of vesiculation with novel microscopy assays to quantify vesiculation dynamics, while genetically tuning protein-protein interactions between the cell wall and outer membrane. Finally, the scope of these studies will be extended to Gram-positive bacteria by determining the dependence of antibiotic resistance on cell wall stiffness in these species, specifically focusing on the mechanical contributions of teichoic acids to resistance. Together, these studies will transform our understanding of bacterial pathogen survival and growth, and point to fresh strategies to circumvent antibiotic resistance and treat bacterial infections.

概括 传统上，抗生素耐药性的研究集中在进化和分子上 电阻机制。但是，我们许多最好的抗生素针对细菌细胞包膜， 这是一种机械稳健的细胞结构外骨骼。最终，这些抗生素原因 细胞死亡通过足够削弱的包络以引起细胞爆炸的大型静水压 其中的压力。尽管细胞信封的机械重要性核心，但我们几乎没有 了解其中哪些分子和部分对承载能力至关重要。 解决这个问题将改变我们对抗生素抗性的理解。主要 我们所知的这一差距的原因是将机械力应用于 单个细菌细胞在监测其生理学时。拟议的研究将解决这个问题 通过应用创新的，高度精确的高通量微流体和基于显微镜的障碍 测量两个主要细胞包膜组件的机械性能的测定 细菌，外膜和细胞壁。这些测定将与分子和 细胞生物学技术和生物物理理论，以探索内部的新兴范式 微生物学：细菌通过自适应调整机械性能来控制抗生素耐药性 它们的细胞信封。首先，建立在最近的地标发现外膜赋予的发现 抗生素对细菌的耐药性由于其机械强度，外部的依赖性 膜刚度和机械抗生素耐药性在细尺度的生化组成上 外膜将被系统地测量。接下来，外膜的机制 囊泡（抗生素耐药性和发病机理的基础过程）将通过 将囊泡的理论机械模型与新的显微镜测定法相结合以量化 囊泡动力学，而细胞壁和 外膜。最后，这些研究的范围将扩展到革兰氏阳性细菌。 确定这些物种中抗生素抗性对细胞壁刚度的依赖性，特别是 侧重于Teichoic Acid对耐药性的机械贡献。这些研究将在一起 改变我们对细菌病原体生存和生长的理解，并指出新的策略 避免抗生素耐药性并治疗细菌感染。