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: 9233901
• 负责人: Jeffrey D. Cirillo
• 金额: $ 37.18万
• 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/9233901
• 关键词: Aftercare; Animal Model; Animals; Attenuated Vaccines; Biological Models; Cause of Death; Cavia; Characteristics; Cleaved cell; Colony-forming units; Communicable Diseases; Custom; Dangerousness; Data; Detection; Development; Disease; Dose; Drug resistance; Drug resistance in tuberculosis; Endoscopy; Enzymes; Evaluation; Fluorescence; Fluorogenic Substrate; Foundations; Gold; Growth; Human; Image; Image Analysis; Imaging technology; In Vitro; Infection; Infectious Agent; Intervention; Lung; Measures; Methods; Monitor; Mus; Mycobacterium tuberculosis; Nature; Organ; Pathogenesis; Physiological; 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; beta-Lactamase; cost; design; efficacy study; fluorescence imaging; head-to-head comparison; imaging study; improved; in vivo imaging; innovation; luminescence; lung imaging; microendoscopy; novel; novel strategies; novel therapeutics; novel vaccines; pathogen; preclinical development; public health relevance; quantitative imaging; respiratory; screening; subcutaneous; therapeutic development; therapeutic evaluation; therapy development; therapy outcome; time use; treatment strategy; tuberculosis treatment; vaccination strategy; vaccine development; vaccine efficacy; vaccine evaluation; whole body imaging

DESCRIPTION (provided by applicant): 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 wil 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.

描述（由申请人提供）：结核病目前困扰着世界近三分之一的人口，但由于病原体结核分枝杆菌的缓慢生长速度以及通过菌落形成单位评估生存力的需要，干预措施的开发进展受到阻碍（菌落形成单位）。拟议的研究旨在通过开发和验证可用于评估结核病新型预防和治疗策略的成像技术来克服这一主要障碍。我们之前的研究已经开发出针对结核病的荧光和生物发光成像策略，但其中最敏感的已被证明是报告酶荧光 (REF)，其在肺部感染期间的检测阈值约为 104 cfu。我们还表明，REF 成像可以在体外和动物治疗后 24-48 小时内进行治疗评估。这项工作为拟议的研究奠定了基础，这些研究旨在允许将 REF 成像应用于治疗效果确定、追踪动物 Mtb 感染以及分析结核病疫苗接种策略。我们已经证明了成像技术能够量化和跟踪肺部和皮下感染，为跟踪结核病疫苗接种和激发剂量提供了独特的机会，这在以前是不可行的，特别是在实时使用活体动物时。在当前的研究中，我们将在早期研究的基础上，测试 REF 成像评估多种治疗方法疗效的能力，提高当前 REF 成像的检测阈值，并证明疫苗功效成像研究的实用性。具体来说，我们建议：1）改进 REF 成像以促进结核病研究。我们的工作假设是，REF 成像的催化性质将使该技术能够达到每个器官 10 cfu 的阈值，足以分析结核病毒力和治疗。我们的初步研究表明，REF 成像非常敏感，成像可用于测量治疗结果，并且有可能改进现有的 REF 底物。为此，我们将比较 CFU 成像的治疗效果评估，分析和改进检测阈值的底物，并仔细评估我们准确追踪 Mtb 感染的能力。 动物模型。 2) 使用成像技术分析疫苗对结核病的功效。我们的工作假设是，显微内窥镜检查和 REF 的结合将使我们能够比传统的基于 CFU 的方法更快、更详细地评估结核疫苗的功效。我们的初步研究表明，我们可以利用显微内窥镜成像来追踪疫苗报告株的皮下接种和 Mtb 肺部感染的 REF 成像。为此，我们将构建稳定的报告疫苗株，并使用它们在小鼠和豚鼠的整个疫苗接种过程中跟踪疫苗接种和攻击剂量，以更好地了解疫苗接种过程中活疫苗和攻击剂量的活力动态。