Target and Non-target Based Resistance to Bedaquiline in South Africa

南非对贝达喹啉的目标和非目标耐药性

• 批准号: 10427170
• 负责人: Ndivhu Makhado
• 金额: $ 34.98万
• 依托单位: SEFAKO MAKGATHO HEALTH SCIENCES UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-20 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10427170
• 关键词: Alabama; Bacillus; Bioenergetics; Biological Assay; Carbon; Cause of Death; Clinical; Clinical Investigator; Collaborations; Complement; Country; Data; Drug Efflux; Drug Targeting; Drug Tolerance; Drug resistance; Drug resistant Mycobacteria Tuberculosis; Electron Transport; Energy Metabolism; Excretory function; Exhibits; Extreme drug resistant tuberculosis; Frequencies; Genes; Glucose; Goals; Health; Health Sciences; Healthcare; Homeostasis; Incidence; Infectious Agent; Intervention; Knowledge; Link; M. tuberculosis genome; Metabolic; Metabolic stress; Metabolism; Multidrug-Resistant Tuberculosis; Mutation; Mycobacterium tuberculosis; Outcome; Oxygen Consumption; Patients; Persons; Pharmaceutical Preparations; Poison; Prevalence; Primary Infection; Process; Regimen; Regulation; Research; Resistance; Respiration; Rifampicin resistance; Role; Science; Series; Shunt Device; South Africa; South African; Succinates; Technology; Testing; Therapeutic; Therapeutic Intervention; Time; Treatment outcome; Tuberculosis; Universities; Variant; Verapamil; base; cohort; drug metabolism; experimental study; extensive drug resistance; extracellular; fitness; flexibility; genome sequencing; glyoxylate; improved; innovation; liquid chromatography mass spectrometry; metabolomics; mortality risk; new technology; novel; pathogenic microbe; preventive intervention; prognostic; programs; prophylactic; stable isotope; transport inhibitor; whole genome

Tuberculosis is the leading cause of death worldwide from a curable infectious agent and is becoming a major concern due to the spread of drug resistant Mycobacterium tuberculosis (Mtb) strains. In 2015, the South African National Tuberculosis Programme introduced Bedaquiline (BDQ) to strengthen existing regimens for the therapy of rifampicin-resistant TB. This initiative has immediate and far-reaching implications for thousands of South Africans suffering from TB. Concerningly, mutations associated with BDQ resistance (rv0678 and atpE) have been identified in Mtb clones isolated from patients who have never been treated with BDQ or clofazimine (CFZ). Intriguingly, both BDQ and CFZ target the Mtb electron transport chain. These findings, together with the fact that Mtb can persist in a dormant, drug-tolerant state, sometimes reactivating to cause TB decades after the primary infection, indicate an urgent need to better understand the mechanisms of BDQ/CFZ resistance in clinical strains of Mtb. Our long-term goal is to understand the mechanisms of Mtb drug resistance and how this knowledge can be used for prophylactic and therapeutic purposes in South Africa. In this proposal, our central hypothesis is that mutations in rv0678, atpE and elsewhere in the Mtb genome dysregulate central metabolism that contributes to BDQ and CFZ resistance. To test this hypothesis, we have established a global collaborative effort between basic and clinical investigators at Sefako Makgatho Health Sciences University (SMU) in South Africa and the University of Alabama at Birmingham (UAB). As part of this collaboration, we have established a series of specific aims to determine the prevalence of rv0678 and atpE mutations in specific Mtb lineages isolated from patients in South Africa. We will also make use of a novel technology termed extracellular flux (XF96) analysis that we have adapted for studying Mtb bioenergetics in real time. This technology will be complemented by 13C stable isotope analyses using liquid chromatography mass spectrometry. Lastly, we will further pursue our exciting preliminary findings and determine whether BDQ resistance associated variants contribute to the bacilli's bioenergetic flexibility. This contribution is significant, because it has the potential to identify a new paradigm that will lead to a mechanistic understanding of the emergence of BDQ/CFZ resistance, and how disruption of this process could be exploited to sterilize Mtb. This proposal is innovative in our opinion, because the newly adapted technology that is supported by whole genome sequencing, real-time bioenergetics and metabolomics, distinguishes itself from conventional approaches for studying drug resistance in pathogenic microbes.

结核病是全球可治愈传染源导致死亡的主要原因，并且正在成为一个主要的疾病 由于耐药结核分枝杆菌 (Mtb) 菌株的传播而引起关注。 2015年，南非 国家结核病规划引入贝达喹啉 (BDQ) 以加强现有治疗方案 耐利福平结核病。这一举措对数千名南方人产生了直接而深远的影响 非洲人患有结核病。令人担忧的是，与 BDQ 耐药性相关的突变（rv0678 和 atpE）已 在从未接受过 BDQ 或氯法齐明 (CFZ) 治疗的患者中分离出 Mtb 克隆。 有趣的是，BDQ 和 CFZ 都以 Mtb 电子传输链为目标。这些发现以及事实 结核分枝杆菌可以持续处于休眠、耐药状态，有时会在发病几十年后重新激活，导致结核病 原发感染，表明临床迫切需要更好地了解 BDQ/CFZ 耐药机制 Mtb 菌株。 我们的长期目标是了解结核分枝杆菌耐药性的机制以及这些知识如何能够 在南非用于预防和治疗目的。在这个提案中，我们的中心假设是 rv0678、atpE 和 Mtb 基因组其他位置的突变会导致中枢代谢失调，从而导致 BDQ 和 CFZ 电阻。为了检验这一假设，我们在全球范围内建立了合作关系 南非 Sefako Makgatho 健康科学大学 (SMU) 的基础和临床研究人员以及 阿拉巴马大学伯明翰分校 (UAB)。作为此次合作的一部分，我们建立了一系列 具体目的是确定从分离的特定 Mtb 谱系中 rv0678 和 atpE 突变的流行率 南非的患者。我们还将利用一种称为细胞外通量 (XF96) 分析的新技术 我们已经适应了实时研究结核分枝杆菌生物能量学。该技术将得到13C的补充 使用液相色谱质谱法进行稳定同位素分析。最后，我们将进一步追求 令人兴奋的初步发现，并确定 BDQ 抗性相关变异是否会导致杆菌的 生物能量的灵活性。 这一贡献意义重大，因为它有可能确定一种新的范式，从而导致 对 BDQ/CFZ 耐药性出现的机制的理解，以及如何破坏这一过程 被用来消灭结核分枝杆菌。我们认为这个提议是创新的，因为新采用的技术 由全基因组测序、实时生物能量学和代谢组学支持，脱颖而出 与研究病原微生物耐药性的传统方法不同。