Minimizing in vivo Drug Tolerance induction in tuberculosis.

最大限度地减少结核病体内药物耐受性的诱导。

基本信息

批准号：
10665033
负责人：
DAVID G RUSSELL
金额：
$ 61.7万
依托单位：
RUTGERS BIOMEDICAL AND HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-23 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10665033
关键词：
Animal Model Animals Antibiotics Bacteria Bacterial Genes Biological Models Biology Bronchoalveolar Lavage CRISPR/Cas technology Cell Line Cell Separation Cell model Cells Clinical Collecting Cell Development Drug Exposure Drug Tolerance Drug resistance Environment Genes Genetic Genetic Databases Genetic Engineering Genome engineering Goals Heritability Heterogeneity Human Immune Infection Link Lung Macrophage Messenger RNA Metabolic Metabolic Pathway Metabolism Modality Modeling Mus Mutation Mycobacterium tuberculosis Outcome Pathway interactions Pharmaceutical Preparations Pharmacotherapy Phenotype Physiology Population Property Regimen Resistance Resources Route Small Interfering RNA Specimen System Systems Biology Technology Testing Therapeutic Therapeutic Intervention Tuberculosis axenic culture candidate identification clinically relevant drug efficacy genetic manipulation global health human disease in vivo network models novel therapeutic intervention pathogen rational design single-cell RNA sequencing success therapeutic siRNA tool transcriptome transcriptome sequencing tuberculosis chemotherapy tuberculosis drugs

项目摘要

Abstract Project 3: Minimizing in vivo drug tolerance induction in tuberculosis. Phenotypic drug tolerance in Mycobacterium tuberculosis is an expression of one of the single most significant properties that make Mtb such a major Global Health threat. Tolerance promotes bacterial persistence in the face of drug therapy and expands the window for emergence of genetically-encoded resistance. The goal of the project is the rational design of a novel therapeutic approach to minimize the induction of drug tolerance through manipulation of the host cells or the immune environment that result in drug tolerance. Such an outcome would enhance the efficacy of current TB drug regimens and reduce emergence of heritable drug resistance. To accomplish this goal, we will leverage a number of recent advances from our labs. The Russell lab has developed the capability to simultaneously profile host and pathogen transcriptomes in cells isolated directly from drug-treated animals. The Sassetti lab has developed a complementary genetic database that comprehensively quantifies the effect of bacterial mutations on drug tolerance during infection, and has exploited CRISPR-Cas9 to engineer the genomes of primary macrophage lines. The overarching hypothesis guiding this project is that phenotypic drug tolerance in Mtb is induced by the host immune environment, which can be specifically modulated to increase drug efficacy. Aim 1: Identify macrophage immune or metabolic pathways that influence Mtb drug tolerance. We will apply different RNA-seq modalities to infected macrophages from the lungs of antibiotic-treated mice to identify host pathways that are linked with the expression of tolerance-related bacterial genes. Integrating diverse Mtb isolates will link these mechanistic observations to clinically-relevant phenotypes. Aim 2: Characterize mechanistic links between macrophage metabolic state and Mtb drug tolerance. We will exploit emergent genetic tools to probe bacterial and host macrophage biology for the functional verification of candidate pathways leading to induction of drug tolerance in Mtb. We will use a combination of culture and host cell model systems to link specific immune pathways to induction of bacterial drug tolerance. Aim 3: Proof-of-Concept for therapeutic interventions to maintain/enhance frontline drug efficacy. We will use synthetic mRNA and siRNA approaches to explore avenues whereby in vivo drug tolerance can be minimized and the efficacy of frontline drugs can be effectively sustained. A combination of reductionist animal models and human clinical specimens will be used to link mechanism with therapeutic relevance.

摘要项目3：在结核病中最大程度地减少体内药物耐受性诱导。结核分枝杆菌中表型药物的耐受性是最重要的一种表达使MTB成为全球重大健康威胁的财产。耐受性促进了细菌的持久性面对药物治疗，并扩大了遗传编码抗性出现的窗口。目标的目标项目是一种新型治疗方法的合理设计，可以最大程度地减少通过操纵宿主细胞或导致药物耐受性的免疫环境。这样的结果会提高当前结核病药物方案的功效并降低可遗传耐药性的出现。为了实现这一目标，我们将利用实验室的最新进展。罗素实验室有发展了直接分离的细胞中同时介绍宿主和病原体转录组的能力来自毒品处理的动物。 Sassetti实验室已经开发了一个互补的遗传数据库全面量化细菌突变对感染过程中药物耐受性的影响，并已开发 CRISPR-CAS9来设计主要巨噬细胞线的基因组。指导该项目的总体假设是MTB中的表型药物耐受性是由宿主免疫环境，可以专门调节以提高药物疗效。目标1：确定影响MTB药物耐受性的巨噬细胞免疫或代谢途径。我们将对来自抗生素处理的小鼠肺的感染巨噬细胞应用不同的RNA-seq模式识别与耐受性相关细菌基因表达相关的宿主途径。整合多样化 MTB分离株将这些机械观察结果与临床上相关的表型联系起来。 AIM 2：表征巨噬细胞代谢状态和MTB药物耐受性之间的机械联系。我们将利用新兴的遗传工具来探测细菌和宿主巨噬细胞生物学的功能验证候选途径，导致MTB诱导药物耐受性。我们将结合培养和宿主细胞模型系统将特定的免疫途径与细菌药物耐受性诱导联系起来。目标3：治疗干预措施的概念证明，以维持/增强前线药物疗效。我们将使用合成mRNA和siRNA方法来探索途径，从而可以使体内药物耐受性是最小化和前线药物的功效可以有效地持续。还原主义动物的结合模型和人类临床标本将用于将机制与治疗相关性联系起来。