Turning Mycobacterium tuberculosis appetite for fatty acids against itself

结核分枝杆菌对脂肪酸的需求与自身相悖

基本信息

批准号：
10592602
负责人：
SABINE EHRT
金额：
$ 21.19万
依托单位：
WEILL MEDICAL COLL OF CORNELL UNIV
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-11-14 至 2024-10-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10592602
关键词：
Acetylcysteine Acute Address Affect Affinity Antibiotic Therapy Antioxidants Antitubercular Antibiotics Bacillus Biology Bone Marrow C3HeB/FeJ Mouse Carbon Catabolism Cell physiology Cholesterol Chronic Clinical Clinical Trials Complement Culture Media Cyclic AMP Cytochrome c Reductase Data Desire for food Diet Doxycycline Drug Targeting Drug Tolerance Electrons Energy Metabolism Environment Enzymes Etiology Fatty Acids Gene Expression Genes Genetic Genetic Determinism Genus Mycobacterium Goals Growth Heat shock proteins Infection Knock-out Knowledge Label Laboratories Lead Lesion Libraries Lipid Peroxidation Lipids Macrophage Malate Synthase Measures Membrane Metabolism Modeling Morbidity - disease rate Mus Mutate Mutation Mycobacterium tuberculosis NADH NADH oxidase Names Oleic Acids Operon Outcome Measure Oxidative Stress Oxidoreductase Oxygen Consumption Pathogenesis Pathogenicity Pathway interactions Persons Pharmaceutical Preparations Phase Phenotype Physiological Physiology Play Predisposition Proteins Protons Reactive Oxygen Species Regimen Research Respiration Respiratory Chain Role Serine Signal Transduction Source Supplementation System Testing Therapeutic Time Toxic effect Treatment Protocols Tuberculosis Work adenosine cyclic-3&apos ,5&apos -monophosphate binding proteins alpha Tocopherol attenuation bactericide caseating granulomas folic acid metabolism host colonization improved inhibitor knock-down long chain fatty acid metabolomics minimal inhibitory concentration mortality mouse model mutant new therapeutic target novel strategies nucleotide metabolism oxidation pathogen prevent response screening transposon sequencing tuberculosis drugs uptake

项目摘要

Project summary Tuberculosis (TB) remains a leading cause of morbidity and mortality worldwide. In 2020, it is estimated that 1.5 million people died from TB. This calls for improved treatment regimens, which will greatly benefit from the identification of novel strategies to sterilize infections. The overarching idea of this line of research is to subvert highly adaptive strategies employed by the etiological agent of TB Mycobacterium tuberculosis (Mtb). There is a large body of work in support of Mtb exploiting host fatty acids as a carbon source. However, although important for growth and survival, fatty acids can also be toxic for Mtb. Previous work from our laboratory showed that mutants devoid of malate synthase (glcB), type-2 NADH dehydrogenase (ndh/ ndhA) and EtfD (etfD) render a fatty acid sensitive phenotype associated with attenuation in the mouse model of TB. This showed that it is possible to turn Mtb’s optimized metabolism to oxidize host fatty acids against the bacilli. Beyond these specific cases, there is a gap in knowledge on the genetic determinants necessary for Mtb to avoid fatty acid toxicity. To address this issue, we have performed transposon sequencing (Tn-seq) and compared transposon mutant libraries generated in medium with and without oleic acid (a long-chain fatty acid commonly used in mycobacteria culture media, and likely a carbon source during infection). This screening identified ndh and etfD transposon mutants as being sensitive to oleic acid, which is consistent with our previous findings. In this proposal we will do a “deep dive” on the physiological function of the top hit of our Tn-seq screening. Preliminary data indicates that an interplay between cAMP signaling, fatty acid catabolism and respiration impacts Mtb drug susceptibility and pathogenicity. Understanding how these different cell processes interact will not only lead to a better understanding of the pathogen’s biology, but it will unveil new strategies to sterilize Mtb infections.

项目摘要结核病（TB）仍然是全球发病率和死亡率的主要原因。在2020年，据估计1.5 百万人死于结核病。这需要改善治疗方案，这将大大受益于鉴定刻板印象的新型策略。这一研究的总体思想是颠覆结核分枝杆菌（MTB）的病因学家采用的高度适应性策略。有一个大量工作以支持MTB利用宿主脂肪酸作为碳源。但是，尽管很重要为了生长和生存，脂肪酸也可能对MTB有毒。我们实验室的先前工作表明没有苹果酸合酶（GLCB），2型NADH脱氢酶（NDH/NDHA）和ETFD（ETFD）的突变体呈现A 脂肪酸敏感的表型与TB小鼠模型中的衰减相关。这表明是可以将MTB优化的代谢转变为氧化物宿主脂肪酸对杆菌的含量。超越这些具体病例，了解MTB所需的遗传决定剂的差距是避免脂肪酸毒性所必需的。到解决这个问题，我们进行了转座子测序（TN-Seq）并比较了转座子突变体在有或没有油酸的培养基中产生的库（分枝杆菌通常使用的长链脂肪酸培养基，可能是感染期间的碳源）。该筛选确定了NDH和ETFD转座子突变体对油酸敏感，这与我们以前的发现一致。在这个建议中，我们将对我们TN-Seq筛选的顶部命中的身体功能进行“深度潜水”。初步数据指示营地信号，脂肪酸分解代谢和呼吸之间的相互作用会影响MTB药物的敏感性和致病性。了解这些不同的细胞过程如何相互作用不仅会导致更好了解病原体的生物学，但它将推出定型MTB感染的新策略。