The role of the anaplerotic node in redox homeostasis and pathogenesis of Mycobacterium tuberculosis and its exploitation as a therapeutic target

回补节点在结核分枝杆菌氧化还原稳态和发病机制中的作用及其作为治疗靶点的开发

• 批准号: BB/T007648/1
• 负责人: Dany Beste
• 金额: $ 72.92万
• 依托单位: University of Surrey
• 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: 未结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FT007648%2F1
• 关键词: role; anaplerotic; node; redox; homeostasis

Summary. The role of the anaplerotic node in redox homeostasis and pathogenesis of Mycobacterium tuberculosis and its exploitation as a therapeutic targetWe desperately need new treatments to control the tuberculosis pandemic, which is fuelled by the emergence of drug-resistant strains of Mycobacterium tuberculosis (Mtb, the causative agent of tuberculosis) as well as a catastrophic synergy with HIV. Mtb is an unusual bacterial pathogen, which has the remarkable ability to cause both acute life threatening disease and also symptomless latent tuberculosis infections that can persist for the lifetime of the human host. It is estimated that 23% of the world has latent tuberculosis, and in 2017 alone, there were 10.0 million cases of tuberculosis and 1.5 million tuberculosis deaths, making TB once again the leading infectious disease globally. A key to the success of tuberculosis as a human disease is ascribed to the extraordinary metabolic flexibility of Mtb. This promotes survival in the variable and harsh environments within the human host, which include exposure to REDuctive and OXidative (redox) stresses. Mtb is able to monitor these stresses in the human host and co-ordinate an appropriate response in order to survive. Maintenance of redox homeostasis is critical to the ability of Mtb to cause disease however many questions in this area of research remain unanswered. A fundamental question is what is the role of metabolism in maintaining redox balance? We have preliminary evidence that a central metabolic hub (the ANA node) consisting of four enzymes is involved in maintaining this balance and have identified a drug which targets this node and kills Mtb. Our previous work showed that this hub is required for the survival of Mtb within its human host cell and we now want to test our hypothesis that this node is essential for the maintenance of redox balance, the ability to cause disease and therefore represents a drug target. We propose to characterise the function of the node in redox control using our mutant strains of Mtb which lack a complete ANA node. Firstly we will investigate the effect of having an incomplete ANA node on the survival, redox balance and immediate metabolic responses of Mtb during exposure to reductive or oxidative stresses. For this work we will use a bespoke system which reports on the redox status of Mtb. The next question is how does the ANA node reconfigure metabolism during redox stress? To tackle this question we will directly analyse the metabolic pathways involved in mediating redox stress using cutting-edge systems-based metabolomics techniques that we have spearheaded at the University of Surrey. By utilising a human cell and mouse model of tuberculosis infection we will probe the essentiality of the ANA node to maintaining redox homeostasis and virulence in the host environment. Finally we will chemically modify a drug which we have shown in preliminary work targets the ANA node and kills Mtb in order to improve the antimicrobial activity. By exploring the mode of action of these tool compounds in the context of redox homeostasis we will validate the ANA node as a drug target. In this way we will elucidate the role of the ANA node in redox control and virulence of this important pathogen and synthesise compounds which can potentially be developed into future treatments for tuberculosis.

总结。我们迫切需要新的治疗方法来控制结核病的大流行，结核病是由结核分枝杆菌耐药菌株（Mtb，结核病的病原体）的出现以及与艾滋病毒的灾难性协同作用推动的。结核分枝杆菌是一种不寻常的细菌病原体，它具有引起急性危及生命的疾病和无症状潜伏性结核感染的非凡能力，这种感染可以在人类宿主的一生中持续存在。据估计，全球23%的人患有潜伏性结核病，仅2017年就有1000万例结核病病例和150万例结核病死亡，结核病再次成为全球主要传染病。结核病作为一种人类疾病取得成功的关键在于结核分枝杆菌非凡的代谢灵活性。这促进了人类宿主在多变和恶劣的环境中生存，包括暴露于还原和氧化（氧化还原）应激。Mtb能够监测人类宿主的这些压力，并协调适当的反应以生存。氧化还原稳态的维持对结核分枝杆菌致病的能力至关重要，但这一研究领域的许多问题仍未得到解决。一个基本的问题是代谢在维持氧化还原平衡中的作用是什么？我们有初步证据表明，一个由四种酶组成的中央代谢枢纽（ANA节点）参与维持这种平衡，并且已经确定了一种靶向该节点并杀死结核分枝杆菌的药物。我们之前的工作表明，这个中心是结核分枝杆菌在其人类宿主细胞内存活所必需的，我们现在想要验证我们的假设，即这个节点对于维持氧化还原平衡至关重要，这是引起疾病的能力，因此代表了一个药物靶点。我们建议使用缺乏完整ANA节点的Mtb突变株来表征该节点在氧化还原控制中的功能。首先，我们将研究在暴露于还原性或氧化应激时，ANA节点不完整对结核分枝杆菌存活、氧化还原平衡和即时代谢反应的影响。对于这项工作，我们将使用一个定制的系统来报告Mtb的氧化还原状态。下一个问题是ANA节点如何在氧化还原应激下重新配置代谢？为了解决这个问题，我们将直接分析参与介导氧化还原应激的代谢途径，使用我们在萨里大学率先采用的基于系统的尖端代谢组学技术。通过利用人类细胞和小鼠结核感染模型，我们将探讨ANA节点在宿主环境中维持氧化还原稳态和毒力的重要性。最后，我们将化学修饰我们在初步工作中展示的靶向ANA节点并杀死Mtb的药物，以提高抗菌活性。通过探索这些工具化合物在氧化还原稳态背景下的作用模式，我们将验证ANA节点作为药物靶点。通过这种方式，我们将阐明ANA节点在氧化还原控制和这一重要病原体的毒力中的作用，并合成可能开发用于结核病未来治疗的化合物。

项目成果

Bedaquiline reprograms central metabolism to reveal glycolytic vulnerability in Mycobacterium tuberculosis.

• DOI: 10.1038/s41467-020-19959-4
• 发表时间: 2020-11-30
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Mackenzie JS;Lamprecht DA;Asmal R;Adamson JH;Borah K;Beste DJV;Lee BS;Pethe K;Rousseau S;Krieger I;Sacchettini JC;Glasgow JN;Steyn AJC
• 通讯作者: Steyn AJC

Murine Norovirus Infection Results in Anti-inflammatory Response Downstream of Amino Acid Depletion in Macrophages.

• DOI: 10.1128/jvi.01134-21
• 发表时间: 2021-09-27
• 期刊: Journal of virology
• 影响因子: 5.4
• 作者: Brocard M;Lu J;Hall B;Borah K;Moller-Levet C;Georgana I;Sorgeloos F;Beste DJV;Goodfellow IG;Locker N
• 通讯作者: Locker N

Structural and Molecular Dynamics of Mycobacterium tuberculosis Malic Enzyme, a Potential Anti-TB Drug Target.

• DOI: 10.1021/acsinfecdis.0c00735
• 发表时间: 2021-01-08
• 期刊: ACS infectious diseases
• 影响因子: 5.3
• 作者: Burley KH;Cuthbert BJ;Basu P;Newcombe J;Irimpan EM;Quechol R;Foik IP;Mobley DL;Beste DJV;Goulding CW
• 通讯作者: Goulding CW

Expanding the boundaries of atomic spectroscopy at the single-cell level: critical review of SP-ICP-MS, LIBS and LA-ICP-MS advances for the elemental analysis of tissues and single cells.

• DOI: 10.1007/s00216-023-04721-8
• 发表时间: 2023-11
• 期刊: ANALYTICAL AND BIOANALYTICAL CHEMISTRY
• 影响因子: 4.3
• 作者: Davison, Claire;Beste, Dany;Bailey, Melanie;Felipe-Sotelo, Monica
• 通讯作者: Felipe-Sotelo, Monica

Structural and molecular dynamics of Mycobacterium tuberculosis malic enzyme, a potential anti-TB drug target

• DOI: 10.1101/2020.07.07.192161
• 发表时间: 2020-07
• 期刊: bioRxiv
• 作者: K. Burley;B. Cuthbert;Piyali Basu;J. Newcombe;E. M. Irimpan;Robert Quechol;Ilona P. Foik;D. Mobley;D. Beste;C. Goulding