PknG mediated tailoring Mycobacterium tuberculosis adaptive metabolism is required for the persister formation

研究综述 结核病已经困扰人类大约7万年了。尽管出现了有效的结核病治疗方法 50多年前的期权，它们既冗长又复杂，并与高频率的 治疗失败。目前结核病治疗的迟钝疗效在很大程度上归因于 结核分枝杆菌(Mtb)形成持续体，这是一小部分耐受于 抗生素的作用。数学模型已经证实了这一点，它表明延长治疗时间 是确保根除持久病毒所必需的。因此，结核分枝杆菌的持久者构成了治疗的关键方面 结核病大流行。然而，人们对结核分枝杆菌形成的潜在代谢过程知之甚少。 坚持在药物治疗中幸存下来的人。 PKng是11种丝氨酸-苏氨酸蛋白激酶之一，它监测和纠正一个受干扰的细胞质氧化还原。 州政府。越来越多的证据表明，有害的活性氧物种(ROS)是由 抗生素和靶细菌代谢。因此，ROS介导的代谢损伤是一种常见的杀微生物剂 效应器。作为对策，结核分枝杆菌已经进化出适应性代谢机制，以规避抗生素- 调节ROS的产生。我们的初步数据证明了这一点，显示结核分枝杆菌试图避免ROS损害， 通过维持必要的代谢活动，如甲基柠檬酸循环(MCC)和TCA循环以维持生存 不是通过使用外部营养，而是通过分解代谢重塑丰富的内源性真菌酸。我们也 观察到pKNG介导的谷氨酰胺-谷氨酸代谢重塑在缓解 过度活化的MCC引起的代谢损伤。总而言之，MCC和/或pKNG介导的失活 调节功能损害持续体的形成。因此，我们假设霉菌酸作为一种内部 碳库，以弥补由于有限的外源碳支持而造成的养分短缺，以及真菌 酸消耗要求pnng和mcc之间存在监管串扰。此应用程序的目标是： 验证霉酚酸消耗(目标1)和pKNG介导的代谢重塑的相对贡献 用于氧化还原动态平衡(目标2)，在周围体形成和随后的药物耐受性。AIMS的结果 1和2将被评估为可用于增强当前标准的疗效的治疗方案 通过防止持久性形成，干扰免疫逃避策略，根除结核病药物治疗方案 代谢同步的结核分枝杆菌(目标3)。我们的工作将提供一种针对串扰的新药物养生法 在Mtb代谢的调节和催化回路之间。这一新选项将提供更简单、更短的 治疗选择将导致提高患者依从性和治愈率，同时减少出现 耐药基因突变。

RESEARCH SUMMARY Tuberculosis (TB) has afflicted humans for roughly 70,000 years. Despite the advent of effective TB treatment options over 50 years ago, they are lengthy and complicated, and are directly associated with high frequency of treatment failure. The blunted efficacy of the current TB treatment is largely attributed to the ability of Mycobacterium tuberculosis (Mtb) to form persisters, a small fraction of phenotypic variants that are tolerant to antibiotic effects. This has been confirmed by mathematical modeling, which showed that prolonged treatment is required to ensure persister eradication. Thus, Mtb persisters constitute a therapeutically critical facet of the TB pandemic. However, little is known regarding the underlying metabolic processes through which Mtb forms persisters in an effort to survive drug treatment. PknG is one of 11 serine-threonine protein kinases and it monitors and corrects a perturbed cytoplasmic redox state. Accumulating evidence suggests that deleterious reactive oxygen species (ROS) are produced by antibiotics and target bacterial metabolism. Thus, ROS-mediated metabolic damage is a common microbicidal effector. As a countermeasure, Mtb has evolved adaptive metabolic mechanisms to circumvent antibiotic- mediated ROS production. Our preliminary data proved this by showing that Mtb seeks to avoid ROS damage, by maintaining essential metabolic activities such as the methylcitrate cycle (MCC) and TCA cycle for survival not by using external nutrients but by catabolically remodeling abundant endogenous mycolic acid. We also observed that PknG-mediated remodeling in glutamine-glutamate metabolism plays a crucial role in mitigating metabolic damage induced by overactive MCC. Taken together, inactivation of the MCC and/or PknG-mediated regulatory function impairs persister formation. Thus, we hypothesize that mycolic acid serves as an internal carbon reservoir to compensate for nutrient shortage due to limited exogenous carbon support, and that mycolic acid consumption requires regulatory crosstalk between PknG and MCC. The goals of this application are: to validate the relative contribution of mycolic acid consumption (Aim 1) and PknG-mediated metabolic remodeling for redox homeostasis (Aim 2) during persister formation and subsequent drug-tolerance. The outcomes of Aims 1 and 2 will be assessed as therapeutic options that can be used to enhance the efficacy of the current standard TB drug regimen by preventing persister formation, interfering with the immune-evasion strategy, and eradicating the metabolically synchronized Mtb (Aim 3). Our work will offer a new drug regimen that targets crosstalk between regulatory and catalytic circuits of Mtb metabolism. This new option will offer simpler and shorter treatment options that will lead to increasing patient compliance and cure rates, while decreasing the emergence of drug-resistant mutations.