Application of Imaging to Development of Tuberculosis Interventions

影像学在结核病干预措施开发中的应用

• 批准号: 8631451
• 负责人: Jeffrey D. Cirillo
• 金额: $ 45.32万
• 依托单位: TEXAS A&M UNIVERSITY HEALTH SCIENCE CTR
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-03-01 至 2019-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8631451
• 关键词: Aftercare; Animal Model; Animals; Attenuated Vaccines; Biological Models; Cause of Death; Cavia; Characteristics; Cleaved cell; Colony-forming units; Communicable Diseases; Custom; Cutaneous; Data; Databases; Detection; Development; Disease; Dose; Drug Resistant Tuberculosis; Drug resistance; Endoscopy; Enzymes; Evaluation; Fluorescence; Fluorogenic Substrate; Foundations; Gold; Growth; Human; Image; Imaging technology; In Vitro; Infection; Infectious Agent; Intervention; Lactamase; Life; Lung; Measures; Methods; Monitor; Mus; Mycobacterium tuberculosis; Nature; Organ; Outcome; Pathogenesis; Population; Prevention strategy; Process; Proteins; Pulmonary Tuberculosis; Recording of previous events; Reporter; Reproducibility; Research; Research Design; Savings; Surface; System; Technology; Testing; Therapeutic; Time; Treatment Efficacy; Tuberculosis; Tuberculosis Vaccines; Vaccination; Vaccines; Validation; Virulence; Work; antimicrobial; base; cost; design; efficacy evaluation; fluorescence imaging; hamstring; head-to-head comparison; improved; in vivo imaging; innovation; luminescence; novel; novel strategies; novel therapeutics; novel vaccines; pathogen; pre-clinical; public health relevance; respiratory; screening; subcutaneous; therapeutic evaluation; therapeutic vaccine; therapy development; time use; treatment strategy; tuberculosis treatment; vaccination strategy; vaccine efficacy; vaccine evaluation; whole body imaging

PROJECT SUMMARY Tuberculosis currently afflicts nearly one-third of the world's population, but progress in development of interventions is hamstrung by the slow growth rate of Mycobacterium tuberculosis, the causative agent, and the need to evaluate viability by colony forming units (cfu). The proposed study is designed to overcome this major roadblock through the development and validation of imaging technologies that can be applied to evaluation of novel prevention and treatment strategies for tuberculosis. Our previous studies have resulted in development of both fluorescent and bioluminescent imaging strategies for tuberculosis, but the most sensitive of these has proven to be reporter enzyme fluorescence (REF), which has a detection threshold of approximately 104 cfu during pulmonary infection. We have also shown that REF imaging allows therapeutic evaluation within 24-48 h post-treatment both in vitro and in animals. This work has set the foundation for the proposed studies designed to allow application of REF imaging to therapeutic efficacy determination, tracking Mtb infections in animals and analysis of vaccination strategies for tuberculosis. We have demonstrated the ability of imaging to quantify and track both pulmonary and subcutaneous infections, offering the unique opportunity to track both vaccination and challenge doses for tuberculosis, which has not been previously feasible, particularly in real-time using live animals. In the current study, we will build upon our earlier studies by testing the ability of REF imaging to evaluate efficacy of a diverse set of therapeutics, improvement of the current REF imaging threshold of detection and demonstrate the utility of imaging study of vaccine efficacy. Specifically, we propose to: 1) Improve REF imaging to facilitate tuberculosis research. Our working hypothesis is that the catalytic nature of REF imaging will allow this technology to achieve thresholds of 10 cfu per organ, sufficient for analysis of tuberculosis virulence and therapeutics. Our preliminary studies demonstrate that REF imaging is very sensitive, imaging can be used to measure therapeutic outcome and that there is potential to improve existing REF substrates. In this aim we will compare therapeutic efficacy evaluation for imaging with cfu, analyze and improve substrates for thresholds of detection and carefully evaluate our ability to accurately track Mtb infection in animal models. 2) Analyze vaccine efficacy against tuberculosis using imaging. Our working hypothesis is that the combination of micro-endoscopy and REF will allow us to evaluate tuberculosis vaccine efficacy more rapidly and in more detail than conventional cfu-based methods. Our preliminary studies demonstrate that we can utilize micro- endoscopic imaging to follow sub-cutaneous inoculation with reporter strains of the vaccine and REF imaging for pulmonary infection with Mtb. In this aim we will construct stable reporter vaccine strains and use them to follow both the vaccination and challenge dose throughout vaccination in mice and guinea pigs to better understand the live vaccine and challenge dose viability dynamics during vaccination.

项目摘要 目前，结核病遭受了世界人口的近三分之一，但发展的进展 干预措施因结核分枝杆菌的生长缓慢，因果剂和 需要通过菌落成型单元（CFU）评估生存能力。拟议的研究旨在克服 通过可以应用于成像技术的开发和验证的主要障碍 评估结核病的新型预防和治疗策略。我们以前的研究已导致 在开发结核病的荧光和生物发光成像策略中，但最多 事实证明，这些敏感是记者酶荧光（参考），其检测阈值 肺部感染期间约有104个CFU。我们还表明，参考成像允许 在体外和动物治疗后24-48小时内的治疗评估。这项工作设定了 旨在允许将REF成像应用于治疗功效的拟议研究基础 确定，跟踪动物中MTB感染以及分析结核病的疫苗接种策略。我们 已经证明了成像量化和跟踪肺和皮下感染的能力， 提供独特的机会来跟踪疫苗接种和挑战剂量的结核病，但尚未 以前是可行的，尤其是在实时使用活动物的实时。在当前的研究中，我们将基于 我们较早的研究通过测试参考成像评估各种治疗剂功效的能力， 改进当前的参考成像检测阈值，并证明了成像研究的实用性 疫苗功效。具体而言，我们建议：1）改进参考成像以促进结核 研究。我们的工作假设是，参考成像的催化性质将使这项技术得以实现 达到每个器官10 CFU的阈值，足以分析结核病的毒力和治疗剂。 我们的初步研究表明，参考成像非常敏感，成像可用于测量 治疗结果，并且有可能改善现有的REF底物。在这个目标中，我们将 比较使用CFU成像的治疗功效评估，分析和改善阈值的底物 检测并仔细评估我们准确跟踪动物模型中MTB感染的能力。 2）分析 使用成像对结核病的疫苗功效。我们的工作假设是 微观镜检查和REF将使我们能够更快地评估结核病疫苗功效 细节比常规的基于CFU的方法。我们的初步研究表明，我们可以利用微型 内窥镜成像以接种疫苗的报告菌株和REF接种后接种后接种 成像用MTB感染肺部。在此目标中，我们将构建稳定的记者疫苗菌株和 在小鼠和豚鼠中，使用它们在整个疫苗接种过程中均跟随疫苗接种和挑战剂量 为了更好地了解疫苗接种期间的实时疫苗并挑战剂量的生存力动力学。