Collateral Consequences of Enabler Genotypes in Antibiotic Treatment Failure.

抗生素治疗失败中促成基因型的附带后果。

• 批准号: 10703351
• 负责人: Jason W. Rosch
• 金额: $ 42.51万
• 依托单位: BROAD INSTITUTE, INC.
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-12 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10703351
• 关键词: Address; Affect; Antibiotic Resistance; Antibiotic Therapy; Antibiotics; Antimicrobial Resistance; Bacterial Infections; Cell physiology; Cells; Chemicals; Clinical; Coal; Development; Evolution; Failure; Genetic; Genetic Transcription; Genotype; Infection; Intermediate resistance; Investigation; Medical; Metabolic; Metabolism; Methodology; Microfluidic Analytical Techniques; Modeling; Morbidity - disease rate; Mutation; Pathway interactions; Patient-Focused Outcomes; Phenotype; Play; Population; Population Analysis; Probability; Production; Regulatory Pathway; Resistance; Role; Serinus; Signal Transduction; Streptococcus pneumoniae; Stress; Therapeutic; Treatment Failure; Treatment outcome; antibiotic tolerance; bacterial metabolism; biological adaptation to stress; genetic manipulation; improved; in vivo; insight; mortality; mutant; prevent; resistance mechanism; response; screening; synergism; tool; treatment optimization

Project 2: Collateral Consequences of Enabler Genotypes in Antibiotic Treatment Failure Antibiotic treatment failure (ATF) is of critical medical importance as delays in effective antibiotic therapy promotes prolonged morbidity and increase mortality leading to overall poorer patient outcomes. While antibiotic resistance is one primary mechanism for ATF, the roles for transient cell-states conferring antibiotic tolerance, persistence, and hetero-resistance, herein referred to as “enablers” are increasingly recognized as a major mechanism for bacterial evasion of antibiotic therapy. These enabler bacterial populations are of considerable scientific and medical importance as typically precede the subsequent evolution or acquisition of high-level resistance mechanisms, making these populations a “canary in the coal mine” for understanding how resistance and treatment failures can emerge. Enabler mutants identified thus far in central cellular pathways indicate commonalities for developing an enabler state suggest these findings will be broadly applicable in a cross-species manner. Our underlying hypothesis is that the collateral phenotypes of enabler genotypes can be identified and rationally targeted to optimize the treatment outcomes and prevent subsequent acquisition of high-level resistance. We plan on using a multifaceted approach to understand the genetic and mechanistic basis of enabler mutations. We will experimentally select for enabler mutations using continuously variable antibiotic exposures in both sensitive strains and those with defined enabler mutations. These antibiotic sensitive but tolerant populations will be used to model the impact of different enabler genotypes on the probability and genetic pathways to subsequent evolution of high-level antibiotic resistance, the role of these genotypes on horizontal transfer of resistance, and how enabler mutants impact antibiotic treatment failure in vivo. These enabler strains will be used to model how the respective genotypes, upon antibiotic exposure, affect key pathways such as bacterial stringent response, transcriptional signaling, and translational fidelity, and how this in turn leads to the production of enabler phenotypes. These lines of investigation will provide insight into whether different enabler genotypes have common mechanistic underpinnings. We also plan to examine if rational targeting of enablers could provide therapeutic benefit to either stop the spread of resistance or improve antibiotic efficacy when enabler mutants are present. Enabler mutations are often associated with collateral consequences that confer sensitivity or resistance to metabolic of chemical stresses. We will determine the possibility of exploiting these cross-sensitivities by both targeted and unbiased chemical screening approaches. These synergies will then be investigated for their capacity to prevent emergence of enabler mutants as well as their capacity to more effectively treat infections caused by enabler strains recalcitrant to antibiotic therapy. The successful completion of these aims will provide a genetic basis and mechanistic insight into transient cell-states conferring enabler phenotypes. We anticipate these findings will be applicable across multiple bacterial species given the likelihood of conserved mechanisms underlying enablers and may provide additional strategies to prevent antibiotic treatment failure during bacterial infection.

项目2：抗生素治疗失败中使能基因型的附带后果 抗生素治疗失败（ATF）是至关重要的医疗重要性，因为延迟有效的抗生素治疗 促进了发病率的延长和死亡率的增加，从而导致总体较差的患者结果。而 抗生素耐药性是ATF的主要机制之一，瞬时细胞状态赋予ATF的作用是， 抗生素耐受性、持久性和异源抗性（本文中称为"使能因子"）正日益受到关注。 被认为是细菌逃避抗生素治疗的主要机制。这些使能细菌 人口具有相当大的科学和医学重要性，因为通常在随后的 进化或获得高水平的抗性机制，使这些种群成为"煤中的金丝雀" 我的"，以了解如何耐药性和治疗失败可以出现。由此鉴定的启动子突变体 在中枢细胞通路中，显示了发展使能状态的共同点，这表明 研究结果将以跨物种方式广泛适用。我们的基本假设是 使能基因型的附属表型可以被鉴定并合理地靶向以优化 治疗结果，并防止随后获得高水平的耐药性。我们计划用一个 多方面的方法来了解使能突变的遗传和机制基础。我们将 实验性地选择使能突变，使用连续可变的抗生素暴露， 敏感菌株和具有确定的使能突变的菌株。这些抗生素敏感但耐受 群体将被用来模拟不同的使能基因型对概率和遗传的影响。 高水平抗生素耐药性的后续进化途径，这些基因型对 耐药性的水平转移，以及使能突变体如何影响体内抗生素治疗失败。这些 使能菌株将被用于模拟各自的基因型如何在抗生素暴露后影响关键的 途径，如细菌的严格反应，转录信号，和翻译的保真度，以及如何 这又导致使能表型的产生。这些调查线索将为我们提供 不同的基因型是否有共同的机制基础。我们还计划 检查是否合理的目标，使能者可以提供治疗效益， 抗性或当使能突变体存在时提高抗生素功效。启动子突变通常 与附带后果有关，这些后果赋予对化学物质代谢的敏感性或抗性， 压力我们将确定利用这些交叉敏感性的可能性， 无偏见的化学筛选方法。然后将调查这些协同作用的能力， 防止使能突变体的出现，以及它们更有效地治疗感染的能力， 通过使能菌株抵抗抗生素治疗。这些目标的成功实现将为 遗传基础和对赋予使能表型的瞬时细胞状态的机制洞察。我们预计 这些发现将适用于多种细菌物种， 可能提供额外的策略，以防止抗生素治疗 在细菌感染期间失败。