Eradicating persistent M. tuberculosis by synthetic lethality of terminal respiratory oxidases

通过终末呼吸氧化酶的合成致死作用根除持续存在的结核分枝杆菌

基本信息

批准号：
10509390
负责人：
Michael Berney
金额：
$ 63.1万
依托单位：
ALBERT EINSTEIN COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-11-13 至 2024-10-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10509390
关键词：
Aerobic Affinity Animal Model Animals Antibiotics Antitubercular Agents Bacillus Bacteria Buffers C3HeB/FeJ Mouse Cells Clinical Combined Modality Therapy Communicable Diseases Complex Cytochrome a Cytochrome bc1 Complex Cytochromes Data Development Disease Progression Dose Drug Kinetics Drug Targeting Drug resistance Drug resistance in tuberculosis Electron Transport Enzymes Goals Granuloma Hallmark Cell Homeostasis Human Hypoxia Immune Immune Evasion Immune response In Vitro Inbred BALB C Mice Individual Infection Investigation Lesion Lung Maintenance Metabolic Metabolic Pathway Modeling Multi-Drug Resistance Mus Mutagenesis Mycobacterium tuberculosis Nutrient Oxidases Oxidation-Reduction Oxidative Phosphorylation Oxidative Stress Oxidoreductase Oxygen Patients Persons Pharmaceutical Preparations Process Property Respiratory Chain Rest Rodent Model Role Structure Structure-Activity Relationship Therapeutic Treatment Protocols Tuberculosis cytochrome c oxidase cytotoxicity drug candidate drug discovery efficacy evaluation fitness test improved in vivo inhibitor knock-down microbicide mouse model mutant mycobacterial necrotic tissue next generation novel novel therapeutics oxidation potency testing prevent pulmonary granuloma resistant strain respiratory response scaffold side effect small molecule inhibitor standard of care stress tolerance stressor synergism transcriptome tuberculosis chemotherapy tuberculosis drugs weapons

项目摘要

Project Summary Mycobacterium tuberculosis (Mtb) kills around 1.8 million people a year, more than any other infectious disease. The two main challenges of combating tuberculosis (TB) are the rapidly increasing number of multidrug-resistant clinical isolates, and the lack of drugs that completely sterilize Mtb infection. The latter is ascribed to the presence of persister cells that are not killed by antibiotics, and also evade the immune response. Mtb persisters are metabolically resting, non-replicating cells that reside in lung granulomas, compact aggregates of immune cells that are the hallmark of tuberculosis. The reduced vasculature of mature granulomas creates a microenvironment low in nutrients and oxygen that induces metabolic quiescence in Mtb. The exact mechanism of the transition to quiescence is unclear, but maintenance of redox homeostasis and oxidative phosphorylation via the electron transport chain appear to be essential to this process. The next generation of anti-TB chemotherapy should be a rational combination of highly active, synergistic drugs that kill both actively dividing cells and persister cells. The FDA-approval of the mycobacterial ATP synthase inhibitor bedaquiline has validated the energy generating machinery of Mtb as a viable drug target. Several new drug candidates (e.g. Q203) that inhibit the cytochrome (Cyt) bc1:aa3 complex, a component of the respiratory chain of Mtb, are in the pipeline. However, all bc1 inhibitors are bacteriostatic in Mtb. The scientific premise of this proposal is that the lack of cidal activity by this class of drugs is due to the presence of a second enzyme, the cytochrome bd oxidase (Cyt-bd). In addition to its role as a terminal oxygen reductase, Cyt-bd is required in cellular redox buffering in response to redox stressors. We hypothesize that (1) combined inhibition of Cyt-bd and Cyt-bc1:aa3 will abrogate terminal oxidation in Mtb, even in granulomas, and (2) inhibition of Cyt-bd will enhance efficacy of front-line and novel drugs to eradicate infection. Our preliminary results show that inactivation of Cyt-bd increases sensitivity to oxidative stress and to standard-of-care anti-TB drugs. We also demonstrated that Mtb lacking Cyt-bd is rapidly killed and cleared in mouse lungs treated with Q203 (Kalia et al. 2017, PNAS). Here, we propose to take the next step towards a new chemotherapeutic approach. In our first aim, we will investigate the synergistic lethality of terminal oxidase inhibition in an animal model that develops granulomatous lesions, similar to human tuberculosis. In aim two, we will evaluate synergies between anti-TB microbicides and inhibition of terminal oxidation. Finally, in aim three, we will focus on developing novel, small- molecule inhibitors of Cyt-bd that synergize with Q203, the pipeline Cyt-bc1:aa3 inhibitor. This will include structure-activity relationship studies, cytotoxicity and pharmacokinetics assessment and in vitro and in vivo potency testing. Successful completion of the proposed studies will contribute to combating TB drug resistance and to developing a sterilizing treatment against TB. !

项目概要结核分枝杆菌 (Mtb) 每年导致约 180 万人死亡，比任何其他传染病都要多疾病。抗击结核病的两个主要挑战是结核病数量的迅速增加临床分离株具有多重耐药性，且缺乏彻底消灭 Mtb 感染的药物。后者是归因于存在未被抗生素杀死并且也逃避免疫的持久细胞回复。结核分枝杆菌持续存在是存在于肺肉芽肿中的代谢静止、非复制细胞，免疫细胞紧密聚集，这是结核病的标志。成熟血管系统减少肉芽肿创造了一个营养和氧气含量低的微环境，导致结核分枝杆菌的代谢静止。过渡到静止的确切机制尚不清楚，但维持氧化还原稳态和通过电子传递链的氧化磷酸化似乎对该过程至关重要。下一代抗结核化疗应该是高效、杀死活跃分裂细胞和持续细胞的协同药物。 FDA批准分枝杆菌 ATP 合酶抑制剂贝达喹啉已验证 Mtb 的能量产生机制是一种可行的药物目标。几种抑制细胞色素 (Cyt) bc1:aa3 复合物的新候选药物（例如 Q203）结核分枝杆菌呼吸链的组成部分正在筹备中。然而，所有 bc1 抑制剂均具有抑菌作用山地车。该提案的科学前提是，此类药物缺乏杀伤活性是由于第二种酶的存在，即细胞色素 bd 氧化酶 (Cyt-bd)。除了作为终端氧气的作用 Cyt-bd 还原酶是细胞氧化还原缓冲中响应氧化还原应激源所必需的。我们假设 (1) Cyt-bd 和 Cyt-bc1:aa3 的联合抑制将消除 Mtb 的末端氧化，甚至在肉芽肿中也是如此，并且 (2)抑制Cyt-bd将增强一线药物和新药根除感染的疗效。我们的初步结果表明，Cyt-bd 失活增加了对氧化应激和标准治疗抗结核病的敏感性药物。我们还证明，缺乏 Cyt-bd 的 Mtb 在用 Q203（Kalia 等人，2017 年，PNAS）。在这里，我们建议采取下一步措施，采用新的化疗方法。在我们的第一个目标中，我们将在开发的动物模型中研究末端氧化酶抑制的协同致死性肉芽肿性病变，类似于人类结核病。在目标二中，我们将评估抗结核病之间的协同作用杀微生物剂和抑制末端氧化。最后，在目标三中，我们将专注于开发新颖的、小型的 Cyt-bd 分子抑制剂与 Q203（管道 Cyt-bc1:aa3 抑制剂）具有协同作用。这将包括构效关系研究、细胞毒性和药代动力学评估以及体外和体内效力测试。成功完成拟议研究将有助于抗击结核病药物耐药性并开发针对结核病的灭菌疗法。！

项目成果

期刊论文数量（6）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Combining CRISPRi and metabolomics for functional annotation of compound libraries.

DOI：
10.1038/s41589-022-00970-3
发表时间：
2022-05
期刊：
Nature chemical biology
影响因子：
14.8
作者：
Anglada-Girotto M;Handschin G;Ortmayr K;Campos AI;Gillet L;Manfredi P;Mulholland CV;Berney M;Jenal U;Picotti P;Zampieri M
通讯作者：
Zampieri M

Bioenergetic Inhibitors: Antibiotic Efficacy and Mechanisms of Action in Mycobacterium tuberculosis.