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 这一综合P01计划依赖于呼吸液滴的生物物理影响M。 结核分枝杆菌(Mtb)的有效载荷、生理和可培养能力，从而影响其传播和感染A 新主人。研究感染的需氧生物学并鉴定有效的细菌基因 ，我们将开发一个易于处理的模拟结核分枝杆菌传播模型，并将其应用于比较传染性。 野生型与突变型结核分枝杆菌菌株，从空洞到终末肺道。鼠标是最多的 广泛使用的气溶胶感染动物模型用于研究结核病、细菌遗传要求 对治疗的反应。有大量的实验证据支持有效的肺部播种 雾化的结核分枝杆菌液滴。我们集团拥有两个大型动物BSL3设施，并拥有长期的 在开发和优化新的动物模型方面的专业知识，以解决需要 翻译工具。 我们还开发了在活动性结核病的改良兔模型中产生空洞干酪的工具，以及各种变化 依赖泡沫巨噬细胞裂解物和粘膜唾液分泌的酪蛋白替代物作为起点 指向。这些矩阵将被优化并用于(I)结核分枝杆菌突变体的体外筛选，以鉴定和验证 结核分枝杆菌在类宿主和环境之间的转换过程中生存所需的候选基因 条件(项目1和2：确定结核分枝杆菌需要的基因和适应性代谢反应 (Ii)作为控制试验中雾化结核分枝杆菌的起始物质 传播给小鼠，以定性和定量地模拟人类呼吸道分泌物的总体特征 生物气溶胶发生器中的动力学，以及(Iii)检验某些药物可能损害传播的假设 尽管在干酪和空洞中积聚的浓度是亚杀菌。 在空气生物学项目4的支持和指导下，我们将结合和利用体外、体外和体内 开发和优化通过受控气溶胶感染模拟传播的小鼠模型的vivo工具。这个 模型将被应用于确认结核分枝杆菌基因和途径-在体外确定-在成功的 从一台主机到另一台主机的多步骤过渡。在小鼠身上表现出丧失传播适合性的突变 将在测试豚鼠的动物间传播方面取得进展(项目3：M。 结核病传播)。 因此，核心B活动服务于每个项目的目标，并将受到航空生物学项目4的密切指导。