Core B: Animal Model Core

核心B：动物模型核心

• 批准号: 10190648
• 负责人: Veronique Dartois
• 金额: $ 51.49万
• 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/10190648
• 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; 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 该综合 P01 计划的前提是呼吸道飞沫的生物物理学会影响 M. 结核病（Mtb）的有效负载、生理学和可培养性，从而影响其传播和感染结核病的能力 研究新宿主的需氧生物学并鉴定有效所需的细菌基因。 传播，我们将开发一个易于处理的模拟 Mtb 传播模型，并将其应用于比较传染性 野生型与突变型 Mtb 菌株，从空腔到终末肺气道，小鼠是最多的。 用于研究结核病、细菌遗传要求的广泛气溶胶感染动物模型 和对治疗的反应 有大量的实验证据支持有效的肺部“播种”。 我们的团队可以使用两个大型动物 BSL3 设施，并拥有长期存在的设施。 开发和优化新动物模型以解决需要的挑战性问题的专业知识 翻译工具。 我们还开发了在改良的活动性结核兔模型中生成空洞干酪的工具，以及变体 依赖于泡沫状巨噬细胞裂解物和粘膜唾液分泌模拟物作为起始的酪蛋白替代物 这些矩阵将被优化并用于 (i) Mtb 突变体的体外筛选以识别和验证。 结核分枝杆菌在类宿主和环境之间的转变过程中生存所需的候选基因 条件（项目 1 和 2：识别 Mtb 所需的基因和代谢适应性反应） 承受传输压力），（ii）作为在受控实验中雾化结核杆菌的起始材料 传播给小鼠，定性和定量地模拟人类呼吸道分泌物的整体特征 生物气溶胶发生器的动力学，以及（iii）检验某些药物可能损害传播的假设 尽管存在空腔，但仍以亚杀菌浓度在酪乳中积聚。 在空气生物学项目 4 的支持和指导下，我们将结合并利用体外、离体和体内 体内工具开发和优化通过受控气溶胶感染模拟传播的小鼠模型。 模型将用于确认 Mtb 基因和途径（体外鉴定）在成功中的贡献 从一种宿主到另一种宿主的多步转变，已证明在小鼠中失去了传播的适应性。 将进一步测试豚鼠的动物间传播（项目 3：马耳他肺炎支原体咳嗽机制） 结核病传播）。 因此，核心 B 活动服务于每个项目的目标，并将受到空气生物学项目 4 的密切指导。