Investigating genetic regulators of Mycobacterium tuberculosis cell division and their role in persistent tuberculosis infection

研究结核分枝杆菌细胞分裂的遗传调节因子及其在持续性结核感染中的作用

• 批准号: 10604773
• 负责人: Frances Marks
• 金额: $ 4.77万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-12-01 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10604773
• 关键词: Actinobacteria class; Acute; Affect; Alanine; Antibiotics; Bacteria; Bacterial Infections; Binding Sites; CRISPR interference; Cell Division Process; Cell Wall; Cell division; Cells; Cellular Structures; Chromosome Segregation; Chronic; Complex; DNA Binding; Data; Defect; Development; Disease model; Drug Targeting; Essential Genes; Gene Expression; Genes; Genetic; Genomics; Goals; Granuloma; Growth; Guide RNA; Homologous Gene; Host Defense; Immune; Immune response; Impairment; In Vitro; Infection; Lead; Macrophage; Methods; Microscopy; Modeling; Morphology; Mus; Mycobacterium tuberculosis; Organism; Pathogenesis; Persons; Phagosomes; Phenotype; Play; Population; Population Heterogeneity; Process; Proliferating; Proteins; Regulation; Regulatory Pathway; Regulon; Replication-Associated Process; Repression; Role; Sagittaria; Stains; Stress; Structure; Technology; Testing; Tetracyclines; Therapeutic; Translating; Tuberculosis; Visualization; Work; adaptive immune response; analog; cellular imaging; daughter cell; gene function; gene repression; genetic technology; human pathogen; improved; in vivo; insight; knock-down; meter; mouse model; mutant; mycobacterial; novel; nutrient deprivation; pathogen; success; transcription factor; tuberculosis drugs; Actinobacteria class; Acute; Affect; Alanine; Antibiotics; Bacteria; Bacterial Infections; Binding Sites; CRISPR interference; Cell Division Process; Cell Wall; Cell division; Cells; Cellular Structures; Chromosome Segregation; Chronic; Complex; DNA Binding; Data; Defect; Development; Disease model; Drug Targeting; Essential Genes; Gene Expression; Genes; Genetic; Genomics; Goals; Granuloma; Growth; Guide RNA; Homologous Gene; Host Defense; Immune; Immune response; Impairment; In Vitro; Infection; Lead; Macrophage; Methods; Microscopy; Modeling; Morphology; Mus; Mycobacterium tuberculosis; Organism; Pathogenesis; Persons; Phagosomes; Phenotype; Play; Population; Population Heterogeneity; Process; Proliferating; Proteins; Regulation; Regulatory Pathway; Regulon; Replication-Associated Process; Repression; Role; Sagittaria; Stains; Stress; Structure; Technology; Testing; Tetracyclines; Therapeutic; Translating; Tuberculosis; Visualization; Work; adaptive immune response; analog; cellular imaging; daughter cell; gene function; gene repression; genetic technology; human pathogen; improved; in vivo; insight; knock-down; meter; mouse model; mutant; mycobacterial; novel; nutrient deprivation; pathogen; success; transcription factor; tuberculosis drugs

PROJECT SUMMARY/ABSTRACT Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis, has infected about one-quarter of the global population and kills over 1 million people every year. Mtb is an exceptionally successful human pathogen largely because the bacteria can adapt to withstand several different stresses in vivo and can enter a state of replicative quiescence where Mtb can persist for months to years. This presents major challenges to treating tuberculosis as many anti-tubercular drugs target bacterial replication and cell wall synthesis. Mtb have a unique mechanism of division where the bacteria grow asymmetrically and produce daughter cells that vary in size, growth rate, and cell wall composition. This contrasts with cell division processes employed by other well-studied bacteria, and Mtb possess distinct cellular machinery and regulatory pathways to coordinate their cell division. Thus, it is critical to understand Mtb’s unique mechanism of replication, how it is regulated, and how it contributes to bacterial pathogenesis. The overall goal of this proposal is to elucidate two novel regulatory networks that control Mtb cell division, and to understand the role this regulation plays during tuberculosis infection when Mtb slow or halt their growth. I hypothesize that the uncharacterized transcription factors WhiA and WhiB2 are essential regulators of cell division in Mtb, and that both proteins are required for maintaining bacterial viability in vivo, even during persistent tuberculosis infection when the bacteria enter a slow or non-replicative state. This proposal seeks to i) utilize novel CRISPR interference technology in Mtb to interrogate the function of two essential and previously uncharacterized genes, ii) define two regulons that are critical for regulating cell division in Mtb, and iii) determine the importance of these regulatory networks during persistent tuberculosis infection. These aims employ phenotypically relevant models of Mtb infection, including macrophage and mouse models of disease. Successful completion of this proposal will advance the field by elucidating how Mtb regulate cell division and provide insight into how these regulatory pathways contribute to Mtb’s persistence and survival in vivo. Understanding mechanisms of Mtb replication and persistence can aid with improving and developing therapeutics for treating tuberculosis.

项目摘要/摘要 结核分枝杆菌（MTB）是结核病的病因，已感染了约四分之一的 全球人口，每年杀死超过100万人。 MTB是一种非常成功的人类病原体 主要是因为细菌可以适应体内几种不同的胁迫，并且可以进入 MTB可以持续数月到几年的复制静止。这给治疗带来了主要挑战 结核病与许多抗结核药物靶向细菌复制和细胞壁合成。 MTB有独特的 细菌不对称生长并产生大小变化的发子细胞的机理， 生长速率和细胞壁组成。这与其他经过充分研究的细胞分裂过程形成对比 细菌和MTB具有不同的细胞机制和调节途径，以协调其细胞分裂。 这是至关重要的，要了解MTB独特的复制机制，如何调节以及如何贡献 到细菌发病机理。该提案的总体目标是阐明两个新型的监管网络 控制MTB细胞分裂，并了解该调节在结核病感染中的作用 缓慢或停止他们的成长。我假设未表征的转录因子和WHIB2是 MTB中细胞分裂的基本调节剂，并且两种蛋白质都是维持细菌生存能力所必需的 在体内，即使在细菌进入缓慢或非复制状态时，即使在持续性结核病感染期间。这 提案寻求i）利用MTB中新型的CRISPR干扰技术来询问两个 必不可少的和以前未表征的基因，ii）定义两种对调节细胞分裂至关重要的法规 在MTB和III中）确定在持续性结核病感染期间这些调节网络的重要性。 这些目的采用了MTB感染的表型相关模型，包括巨噬细胞和小鼠模型 疾病。成功完成该提案将通过阐明MTB调节细胞来推进该领域 分裂并提供有关这些监管途径如何促进MTB的持久性和生存的见解 体内。了解MTB复制和持久性的机制可以帮助改善和发展 治疗结核病的治疗剂。