Core B: Animal Model Core

核心B：动物模型核心

• 批准号: 10404529
• 负责人: Veronique Dartois
• 金额: $ 46.89万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-13 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10404529
• 关键词: Address; Aerosols; Affect; Animal Model; Animals; Automobile Driving; Bacillus; Bacteria; Bacterial Genes; Bacteriology; Bar Codes; Biological; Biophysics; Candidate Disease Gene; Cavia; Clinical; Coughing; Data Set; Devices; Disease; Dose; Doxycycline; Exposure to; Foamy Macrophage; Generations; Genes; Genetic; Genetic study; Genomic DNA; Histologic; Human; Impairment; In Vitro; Infection; Liquid substance; Lung; Measures; Metabolic; Metabolic Pathway; Modeling; Mus; Mycobacterium tuberculosis; Nose; Oryctolagus cuniculus; Outcome; Output; Pathway interactions; Pharmaceutical Preparations; Physiology; Program Research Project Grants; Publishing; Reproducibility; Resource Sharing; Stress; Testing; Time; Validation; Variant; Viscosity; Work; aerosolized; bacterial genetics; bactericide; biosafety level 3 facility; clinically relevant; deep sequencing; experimental study; fitness; fly; immunopathology; in vivo; knock-down; man; mouse model; mutant; programs; respiratory; response; tool; transmission process; treatment response; tuberculosis drugs

Shared Resource Core B – Abstract This integrated P01 program relies on the premise that the biophysics of respiratory droplets impacts M. tuberculosis (Mtb) payload, physiology and culturability, and thus influences its ability to transmit and infect a new host. To study the aerobiology of infection and identify the bacterial genes required for effective transmission, we will develop a tractable model of simulated Mtb transmission and apply it to compare infectivity of wild type versus mutant Mtb strains, from cavity caseum to terminal lung airways. The mouse is the most widespread animal model of aerosol infection used to investigate TB disease, bacterial genetic requirements and response to therapy. There is extensive experimental evidence supporting efficient lung ‘seeding’ by aerosolized Mtb droplets. Our group has access to two large animal BSL3 facilities and has a long-standing expertise in developing and optimizing new animal models to address challenging questions that require translational tools. We have also developed tools to generate cavity caseum in a modified rabbit model of active TB, and variations of caseum surrogate that rely on foamy macrophage lysate and mucosalivary secretion mimics as the starting point. These matrices will be optimized and used (i) in in vitro screens of Mtb mutants to identify and validate candidate genes required for survival of Mtb during the transitions between host-like and environmental conditions (with Projects 1 and 2: identification of genes and adaptive metabolic responses that Mtb requires to survive transmission stresses), (ii) as the starting material to aerosolize Mtb bacilli in experiments of controlled transmission to mice, to qualitatively and quantitively mimic overall features of human respiratory secretion dynamics in the bioaerosol generator, and (iii) to test the hypothesis that some drugs may impair transmission despite accumulating in caseum and cavities at concentrations that are sub-bactericidal. With support and guidance from the Aerobiology Project 4, we will combine and leverage in vitro, ex vivo and in vivo tools to develop and optimize a mouse model of simulated transmission by controlled aerosol infection. The model will be applied to confirm the contribution of Mtb genes and pathways – identified in vitro – in the successful multi-step transition from one host to another. Mutants with demonstrated loss of fitness for transmission in mice will progress to testing between-animal transmission in guinea pigs (Project 3: Mechanisms of cough in M. tuberculosis transmission). Thus Core B activities serve the objectives of each project and will be closely guided by Aerobiology Project 4.

共享资源核心B - 摘要 该集成的P01程序依赖于这样的前提：呼吸液滴的生物物理会影响M。 结核病（MTB）有效载荷，生理和文化，因此影响其传播和感染的能力 新主机。研究感染的环境生物学并确定有效的细菌基因 传输，我们将开发一个可访问的模拟MTB传输模型，并将其应用以比较感染 野生型与突变体MTB菌株，从腔caseum到末端肺气道。鼠标最多 用于研究结核病疾病的气溶胶感染的宽度动物模型，细菌遗传需求 和对治疗的反应。有大量的实验证据支持有效的肺“种子” 雾化的MTB液滴。我们的小组可以使用两个大型动物BSL3设施，并且有一个长期的 开发和优化新动物模型以解决需要的挑战问题的专业知识 翻译工具。 我们还开发了工具以在有效TB的修改后的兔子模型中生成空腔Caseum，并变化 依靠泡沫巨噬细胞裂解物和粘膜分泌物模仿的Caseum代理 观点。这些矩阵将被优化和使用（i）MTB突变体的体外筛选以识别和验证 MTB生存所需的候选基因在宿主样和环境之间的过渡期间 条件（带有项目1和2：MTB需要的基因和自适应代谢反应的识别 在受控的实验中存活传播应力），（ii）作为雾化MTB杆菌的起始材料 向小鼠传播，以定性和定量模仿人呼吸道分泌的整体特征 生物Aerosol发生器中的动力学，以及（iii）测试某些药物可能会损害传播的假设 在凯西姆和腔体中积累的任务，以亚细菌的浓度为浓度。 借助Aerobiology项目4的支持和指导，我们将在体外，Ex Vivo和In结合并利用 通过受控气溶胶感染开发和优化模拟传播的鼠标模型的Vivo工具。这 模型将用于确认MTB基因和途径的贡献（在体外鉴定） 从一个主机到另一个主机的多步过渡。突变体显示出小鼠传播的适应性丧失 将进展为测试豚鼠的动物间传播（项目3：咳嗽的机制。 结核病传播）。 该核心B活动为每个项目的目标提供服务，并将受到特技生物学项目4的密切指导。