A single cell, single molecule microscopy platform for antibiotics research

用于抗生素研究的单细胞、单分子显微镜平台

• 批准号: BB/T017570/1
• 负责人: Seamus Holden
• 金额: $ 51.34万
• 依托单位: Newcastle University
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2020
• 资助国家: 英国
• 起止时间: 2020 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FT017570%2F1
• 关键词: single; cell; molecule; microscopy; platform

The use of antibiotics to supress and treat bacterial infections is a cornerstone of modern medicine. However, bacteria are increasingly developing resistance towards multiple classes of antibiotics currently in clinical use. Some strains of life-threatening bacteria like tuberculosis are already resistant to all available antibiotics, making them effectively untreatable. The rapidly unfolding antibiotic resistance crisis has been declared a global health emergency by the World Health Organization, and research aiming to tackle antibiotic resistance a strategic priority both by UK Government and UKRI. To address this major threat to human health, we need to ramp up our efforts to screen and develop novel antibiotics that can be used against the multidrug resistant bacteria already in circulation, and also to develop approaches to re-sensitise them for already existing antibiotics. In the longer term, it is crucial to identify novel antibiotic targets and treatment strategies with an intrinsically reduced risk of resistance development. In recent years, bacteria have been found to have a highly complex and dynamic cellular internal organisation. As a result, many of the central properties of antibiotics such as their ability to trigger cell rupture can only be understood in the cellular framework. Following the cellular consequences of a novel antibacterial compound via microscopy is thus an extremely powerful tool to understand how antibiotics work (mode of action). Bacteria also exhibit large cell-to-cell differences across a single population that allows individual bacteria to evade and resist antibiotics. Due to the single cell nature of these phenomena, microscopic techniques are essential for understanding how antibiotics interact with bacterial cells and populations. The Newcastle University Centre for Bacterial Cell Biology (CBCB) is world leading in studying the structure and function of bacterial cells. However, our research focus is not limited to fundamental bacterial cellular biology. Through research on host-pathogen interactions, antibiotic mode of action, identification of novel antibiotic targets, and also through direct novel antibiotic screening projects, researchers at the CBCB are actively engaged in research that aims to translate the gained knowledge to novel antibiotic discoveries and therapies. In the very core of the success of CBCB has been a suite of high performance microscopes dedicated and optimised for work with live bacteria, including pathogenic ones. However, microscopy is still a rapidly developing field with new instrumentation enabling approaches that were previously not feasible. We have identified three complementary, cutting-edge techniques that we foresee to become particularly important for antibiotics research: (i) image-based screening for novel antibiotics, (ii) single cell imaging combined with on-chip drug treatment to understand how antibiotics kill bacteria and how bacteria resist antibiotics, (iii) single molecule microscopy that allows antibiotic action to be monitored directly on the level of individual proteins and complexes. We request funds for the purchase of a microscope system capable of all three techniques, which thus is both extremely powerful and provides excellent value for money. Advanced microscopy systems such as this one require a high level of expertise that often prevents effective adoption of these techniques by non-specialists. To deliver access to these techniques to a widest user base, Newcastle University is supporting this application with commitment to hire a PhD-level staff scientist dedicated to assisting users with experimental planning, image acquisition and image analysis, in addition to managing access and maintenance.

使用抗生素来抑制和治疗细菌感染是现代医学的基石。然而，细菌对目前临床使用的多种抗生素的耐药性越来越强。一些威胁生命的细菌菌株，如结核病，已经对所有可用的抗生素产生耐药性，使其无法有效治疗。世界卫生组织已宣布迅速展开的抗生素耐药性危机为全球卫生紧急情况，旨在解决抗生素耐药性的研究是英国政府和UKRI的战略重点。为了应对这一对人类健康的重大威胁，我们需要加大力度筛选和开发可用于对抗已在流通中的多重耐药细菌的新型抗生素，并开发使其对现有抗生素重新敏感的方法。从长远来看，确定新的抗生素靶点和治疗策略至关重要，其本质上降低了耐药性发展的风险。近年来，人们发现细菌具有高度复杂和动态的细胞内部组织。因此，抗生素的许多核心特性，例如它们触发细胞破裂的能力，只能在细胞框架中理解。因此，通过显微镜观察新型抗菌化合物的细胞后果是了解抗生素如何工作（作用模式）的一个非常强大的工具。细菌在单个种群中也表现出很大的细胞间差异，这使得单个细菌能够逃避和抵抗抗生素。由于这些现象的单细胞性质，显微技术对于了解抗生素如何与细菌细胞和种群相互作用至关重要。纽卡斯尔大学细菌细胞生物学中心（CBCB）在研究细菌细胞的结构和功能方面处于世界领先地位。然而，我们的研究重点并不局限于基本的细菌细胞生物学。通过对宿主-病原体相互作用、抗生素作用模式、新型抗生素靶点的鉴定以及直接的新型抗生素筛选项目的研究，CBCB的研究人员积极参与旨在将所获得的知识转化为新型抗生素发现和疗法的研究。CBCB成功的核心是一套高性能显微镜，专门用于活细菌（包括致病菌）的工作并进行了优化。然而，显微镜仍然是一个快速发展的领域，新的仪器使以前不可行的方法成为可能。我们已经确定了三种互补的尖端技术，我们预计这些技术对抗生素研究特别重要：（i）基于图像的新型抗生素筛选，（ii）单细胞成像与芯片上药物治疗相结合，以了解抗生素如何杀死细菌以及细菌如何抵抗抗生素，（iii）单分子显微镜，其允许在单个蛋白质和复合物的水平上直接监测抗生素作用。我们要求提供资金，用于购买一个能够使用所有三种技术的显微镜系统，该系统功能非常强大，性价比极高。像这样的先进显微镜系统需要高水平的专业知识，这往往会阻碍非专业人士有效地采用这些技术。为了向最广泛的用户群提供对这些技术的访问，纽卡斯尔大学正在支持这一应用，并承诺聘请一名博士级科学家，致力于协助用户进行实验规划、图像采集和图像分析，以及管理访问和维护。

项目成果

Molecular motor tug-of-war regulates elongasome cell wall synthesis dynamics in Bacillus subtilis

分子运动拔河比赛调节枯草芽孢杆菌细胞壁合成动力学

• DOI: 10.1101/2023.05.10.540107
• 发表时间: 2023
• 作者: Middlemiss S

The structural basis of hyperpromiscuity in a core combinatorial network of type II toxin-antitoxin and related phage defense systems.

• DOI: 10.1073/pnas.2305393120
• 发表时间: 2023-08-15
• 期刊: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• 影响因子: 11.1
• 作者: Ernits, Karin;Saha, Chayan Kumar;Brodiazhenko, Tetiana;Chouhan, Bhanu;Shenoy, Aditi;Buttress, Jessica A.;Duque-Pedraza, Julian J.;Bojar, Veda;Nakamoto, Jose A.;Kurata, Tatsuaki;Egorov, Artyom A.;Shyrokova, Lena;Johansson, Marcus J. O.;Mets, Toomas;Rustamova, Aytan;Dzigurski, Jelisaveta;Tenson, Tanel;Garcia-Pino, Abel;Strahl, Henrik;Elofsson, Arne;Hauryliuk, Vasili;Atkinson, Gemma C.
• 通讯作者: Atkinson, Gemma C.