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.

研究综述 抗生素未能控制细菌性疾病，通常是由于耐药（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，因为它能够 产生耐药突变体来抵抗抗生素的作用。治疗耐药结核病（TB）患者需要两年时间， 每人花费近393，000美元，这比每人治疗约49，000美元要贵得多 一名药物敏感（DS）结核病患者。尽管人类健康问题迫在眉睫，但人们对人类的健康状况知之甚少。 耐药结核病发展的机制基础。鉴于基因组突变率低， 结核分枝杆菌复制，内在细菌因素应该在发展耐药结核病中发挥重要作用，但它们 被研究了。越来越多的证据表明，结核分枝杆菌持续存在的周期性形成， 对结核抗生素产生短暂耐受的变异，可使结核患者易于出现永久性DR 变种人我们最近报道了结核分枝杆菌持续者的非靶向代谢组学分析，并揭示了结核分枝杆菌转移 其海藻糖介导的碳通量朝向中心碳代谢（CCM）中间体的生物合成 以避免不可逆的抗生素损伤，同时减少其流向细胞壁分枝菌酰生物合成的通量 糖脂这一过程被称为“海藻糖催化转换”，并被确定为结核分枝杆菌所必需的 持久性形成、存活力和药物耐受性。在本申请中，我们假设海藻糖催化 转移是Mtb在用TB抗生素治疗后执行的适应性策略，以实现药物耐受性， 也促进DR突变体的发展，从而改变TB病程。在横断面研究中， 与7个不同的结核病临床谱系，2个谱系如HN 878 W-北京株（HN 878）， 与发展成多药耐药（MDR）-TB的高风险和高死亡率相关。因此，我们将研究 我们的假设是通过证明HN 878是高毒力的，比HN 878更容易产生耐药性， 其他谱系菌株，因为其高水平的海藻糖催化转化活性。为此，我们将确定， 海藻糖催化转换是HN 878的内在因子，负责其药物耐受性、DR突变率， 体外、离体高毒力，然后使用TB鼠模型将其应用于体内。成功的结果 应用将有助于开发新的治疗干预措施，治愈耐药结核病患者，包括那些 感染了HN 878。