Interplay of M. tuberculosis trehalose metabolism and its pathogenesis and drug resistance

结核分枝杆菌海藻糖代谢及其发病机制和耐药性的相互作用

• 批准号: 10585346
• 负责人: Hyungjin Eoh
• 金额: $ 66.19万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-16 至 2027-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10585346
• 关键词: Aftercare; Anabolism; Antibiotics; Antitubercular Antibiotics; Bacillus; Bacterial Infections; C3HeB/FeJ Mouse; CRISPR interference; Carbon; Cell Wall; Clinical; Cord Factors; Cross-Sectional Studies; Development; Disaccharides; Disease; Drug Targeting; Drug Tolerance; Drug resistance; Drug resistance in tuberculosis; Dyes; Exhibits; Flow Cytometry; Genomics; Glycolipids; Goals; Health; Heterogeneity; Human; Immune system; In Vitro; Infection; Intrinsic factor; Label; Link; Mediating; Messenger RNA; Metabolic; Metabolism; Microbial Biofilms; Multidrug-Resistant Tuberculosis; Mutation; Mycobacterium tuberculosis; Outcome; Pathogenesis; Patients; Pattern; Periodicity; Persons; Phenotype; Play; Population; Process; Public Health; Quantitative Reverse Transcriptase PCR; Regimen; Reporting; Research; Role; Source; Testing; Therapeutic; Therapeutic Intervention; Treatment Protocols; Trehalose; Tuberculosis; Variant; Virulence; Virulence Factors; Work; aged; antibiotic tolerance; bactericide; base; cost; drug development; drug resistance development; drug-sensitive; epidemiology study; high risk; in vivo; inhibitor; insight; metabolomics; mortality; mouse model; mutant; novel therapeutic intervention; novel therapeutics; overexpression; pathogen; resistance mutation; synergism; tuberculosis drugs; tuberculosis treatment; validamycins

Research Summary Antibiotics have failed to control bacterial diseases typically due to the emergence of drug resistant (DR) mutants. Mycobacterium tuberculosis (Mtb) is one of the world’s most successful pathogens because of its capacity to develop DR mutants to withstand antibiotic effects. Treating DR-tuberculosis (TB) patients takes two years and costs nearly $393,000 per person, which is substantially more expensive than ~ $49,000 per person for treating a drug sensitive (DS)-TB patient. Despite this pressing human health problem, little is known about the mechanistic bases underlying the development of DR-TB. Given the low genomic mutation rates and slow replication of Mtb, intrinsic bacterial factors should play an important role in developing DR-TB, but they have been understudied. Accumulating evidence has shown that cyclic formation of Mtb persisters, a phenotypic variant transiently tolerant to TB antibiotics, can predispose TB patients to the emergence of permanent DR mutants. We recently reported untargeted metabolomics profiling of Mtb persisters and revealed that Mtb shifted its trehalose-mediated carbon flux towards the biosynthesis of central carbon metabolism (CCM) intermediates to avoid irreversible antibiotic damage, while decreasing its flux towards the biosynthesis of cell wall mycolyl glycolipids. This process was termed the “trehalose catalytic shift” and was identified to be essential for Mtb persister formation, viability, and drug tolerance. In this application, we hypothesize that the trehalose catalytic shift is an adaptive strategy executed by Mtb after treatment with TB antibiotics to achieve drug tolerance and also to facilitate the development of DR mutants, thus altering the TB disease course. In cross-sectional studies with 7 different clinical TB lineages, lineage 2 strains such as HN878 W-Beijing strain (HN878), have been associated with a high risk of developing multidrug resistant (MDR)-TB and high mortality. Thus, we will examine our hypothesis by demonstrating that HN878 is hypervirulent and more prone to develop drug resistance than other lineage strains because of its high level of trehalose catalytic shift activity. To this end, we will determine if the trehalose catalytic shift is an HN878 intrinsic factor responsible for its drug tolerance, DR mutation rates, and hypervirulence in vitro, ex vivo and then apply it in vivo using a TB murine model. A successful outcome of this application will aid in the development of new therapeutic interventions to cure DR-TB patients, including those infected with HN878.

研究摘要 抗生素未能控制细菌疾病，通常是由于耐药性（DR）突变体的出现。 结核分枝杆菌（MTB）是世界上最成功的病原体之一，因为它的能力 开发突变体以承受抗生素作用。治疗dr-berculculosis（TB）患者需要两年 每人的费用将近393,000美元，这比每人$ 49,000的昂贵 药物敏感（DS）-TB患者。尽管有这种紧迫的人类健康问题，但对 DR-TB开发的基础机械基础。考虑到低基因组突变率和缓慢 MTB，固有细菌因素的复制应在开发DR-TB中起重要作用，但它们具有 被理解。积累的证据表明，MTB持久的循环形成，一种表型 变体对结核病抗生素的变体耐受性，可以使结核病患者容易出现永久性DR 突变体。我们最近报道了MTB Persister的不靶向代谢组学分析，并透露MTB发生了变化 其海藻糖介导的碳通量向中央碳代谢（CCM）中间体的生物合成 为了避免不可逆的抗生素损伤，同时减小了细胞壁霉菌基因合成的通量 糖脂。该过程被称为“海藻糖催化转移”，被确定为MTB必不可少的 迫害形成，生存力和药物耐受性。在此应用中，我们假设海藻糖催化 转移是用结核病抗生素治疗后MTB执行的一种自适应策略，以实现药物耐受性和 还促进了突变体博士的发展，从而改变了结核病病程。在横断面研究中 使用7种不同的临床结核谱系，谱系2菌株，例如HN878 W-Beijing菌株（HN878） 与产生耐多药（MDR）-TB和高死亡率的高风险有关。那我们将检查 我们的假设是通过证明HN878高度呼吸的，并且比以前更容易产生耐药性 由于其高水平的海藻糖催化偏移活性，其他谱系菌株。为此，我们将确定是否 海藻糖催化转移是HN878的内在因素，负责其药物耐受性，DR突变率和 在体外过度投资，离体外，然后使用TB鼠模型在体内施加。成功的结果 应用将有助于开发新的治疗干预措施以治愈DR-TB患者，包括 感染了HN878。