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

研究总结 抗生素未能控制细菌疾病，通常是由于耐药(DR)突变的出现。 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 开发耐受抗生素影响的DR突变体。治疗耐药结核病(TB)患者需要两年时间和 每人花费近393,000美元，这远远高于每人49,000美元的治疗费用 一名对药物敏感(DS)的结核病患者。尽管这是一个紧迫的人类健康问题，但人们对 耐药结核病发生的机制基础。考虑到基因组突变率低和缓慢 结核分枝杆菌的复制，内在的细菌因素应该在DR-TB的发展中发挥重要作用，但它们已经 一直未得到充分研究。越来越多的证据表明，结核分枝杆菌的循环形成持续存在，这是一种表型 对结核抗生素短暂耐受的变种，可使结核病患者易出现永久性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特征，并揭示了结核分枝杆菌移位 其海藻糖介导的碳流向中心碳代谢(CCM)中间产物的生物合成 以避免不可逆转的抗生素损伤，同时减少其向细胞壁真菌生物合成的通量 糖脂。这一过程被称为“海藻糖催化转移”，被认为是结核分枝杆菌所必需的。 持久性形成、生存能力和药物耐受性。在这个应用中，我们假设海藻糖催化 SHIFT是结核分枝杆菌在使用抗生素治疗后执行的一种适应性策略，以实现药物耐受和 也是为了促进DR突变体的发展，从而改变结核病的病程。在横断面研究中 与7个不同的临床结核病谱系，谱系2株，如HN878W-北京株(HN878)，已被 与发展多药耐药(MDR)结核病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HN878超强毒力和更容易产生耐药性 其他谱系菌株由于其高水平的海藻糖催化转移活性。为此，我们将确定是否 海藻糖催化移位是HN878的内在因素，与其耐药、DR突变率和 体外超强毒力，体外应用，然后在体内应用结核小鼠模型。这是一个成功的结果 应用将有助于开发新的治疗干预措施来治愈DR-TB患者，包括 感染了HN878。

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.