Direct quantitation of the circulating Mtb-peptidome for pediatric TB management

直接定量循环 Mtb 肽组用于儿科结核病管理

• 批准号: 9333558
• 负责人: Tony Y. Hu
• 金额: $ 44.05万
• 依托单位: ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-03-17 至 2022-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9333558
• 关键词: Address; Adult; Adverse effects; Antibodies; Antigens; Antitubercular Agents; Area; Bacillus (bacterium); Bacteria; Bacteriology; Biological Assay; Biological Markers; Blood; Blood Volume; Blood specimen; CXCL10 gene; Chest; Child; Childhood; Clinical; Clinical/Radiologic; Custom; DNA; Data; Detection; Diagnosis; Diagnostic; Diagnostic Procedure; Diagnostic Sensitivity; Diagnostic Specificity; Disease; Disease Progression; Early Diagnosis; Enrollment; Evaluation; Exposure to; Extreme drug resistant tuberculosis; Growth; HIV; Health; Hour; Immune response; Immunologics; Infection; Investigation; Longitudinal Studies; Mass Spectrum Analysis; Measures; Methods; Microscopy; Modeling; Molecular Weight; Monitor; Morbidity - disease rate; Mycobacterium Infections; Mycobacterium tuberculosis; National Institute of Child Health and Human Development; Noise; Organism; Patients; Pattern; Peptide Signal Sequences; Peptides; Performance; Physicians; Population; Prospective cohort; Proteins; Proteomics; Publishing; Reaction Time; Recruitment Activity; Regimen; Research Infrastructure; Resources; Retrospective cohort; Sampling; Sensitivity and Specificity; Serum; Severities; Severity of illness; Signal Transduction; South Africa; Specificity; System; Techniques; Technology; Time; Treatment Efficacy; Treatment Protocols; Treatment outcome; Trypsin; Tuberculosis; Validation; Work; accurate diagnosis; base; biobank; clinical Diagnosis; clinically relevant; co-infection; cohort; extensive drug resistance; high risk; improved; improved outcome; interest; miniaturize; mortality; mycobacterial; nanoparticle; nanoshell; non-tuberculosis mycobacteria; novel; pediatric patients; predict clinical outcome; profiles in patients; prospective; proteomic signature; response; sample collection; titanium dioxide; treatment duration; treatment response; tuberculosis drugs; tuberculosis treatment

ABSTRACT Conventional methods for adult and pediatric tuberculosis (TB) diagnosis and treatment monitoring rely heavily on time-consuming bacterial culture or unquantifiable DNA assays to detect the presence of small numbers of bacteria. Pediatric TB management is particularly difficult because current organism-based methods for diagnosis and treatment monitoring are resource-intensive, invasive and inadequately sensitive for children, who tend to have paucibacillary disease. Due their immunological immaturity, young children are at high risk of rapid disease TB progression, with significant morbidity and mortality. Assessment of their treatment response relies on subjective measures, limiting children's access to much needed shorter and safer TB regimens. More rapid and rigorous methods of TB diagnosis and treatment monitoring are thus critically needed for children. In order to address current limitations in pediatric TB management, we propose to develop a rapid diagnostic method, independent of mycobacterial isolation, that quantifies the low molecular-weight M. tuberculosis (Mtb) antigens (CFP-10 and ESAT-6) and the TB-associated host marker IP-10 in patient blood samples. ESAT-6 and CFP-10 are ideal biomarkers to detect active TB and its treatment response, and addition of IP-10 to this Mtb-specific proteomic pattern will provide an integrated profile of patients' pathophysiological changes during anti-TB treatment, while also improving diagnostic sensitivity. Our strategy utilizes hollow, energy-focusing TiO2 NanoShells functionalized with custom antibodies specific for Mtb CFP-10 and ESAT-6 and host IP-10 peptides with high-throughput mass spectrometry (MS) to increase diagnostic sensitivity and specificity. We evaluated an Mtb-antigen-only version of this NanoShell-MS platform (no anti-IP-10 NanoShells) with 292 adult and 102 pediatric patients and controls chosen from five highly relevant cohorts (adult, pediatric and latent TB, HIV/TB co-infection, and non-TB mycobacterial infection), from our global collaborators. NanoShell-MS sensitivity and specificity for active TB (bacteriologically confirmed and clinically diagnosed) were, respectively, 90.7% / 97.7% in adults and 88.2% / 100% in children, with 78.6% sensitivity in culture-negative pediatric TB patients, greatly exceeding the WHO's published 4-15% Xpert sensitivity in this cohort. NanoShell-MS results can be obtained within one hour of sample collection compared to 4-6 weeks for conventional culture, and detected decreases in blood Mtb antigen levels within four days of anti-TB treatment initiation. In this proposal we aim to 1) use NanoShell-MS profiling to develop a quantitative prediction model for active TB diagnosis in large, well-described prospective pediatric TB cohorts and 2) determine the utility of our approach for rapid evaluation of treatment efficacy. Our NanoShell-MS assay platform has the advantage that it uses accurate, high-throughput mass spectrometry, which has become a critical means for clinical diagnosis in many parts of the world, while integration of our NanoShell-MS platform with an easy-to-use “miniaturized” (shoebox size) MS system which should also allow this approach to serve pediatric patients in resource-limited areas.

抽象的 成人和儿童结核病 (TB) 诊断和治疗监测的传统方法严重依赖 耗时的细菌培养或无法定量的 DNA 检测来检测少量细菌的存在 儿童结核病的管理尤其困难，因为目前基于微生物的方法。 诊断和治疗监测是资源密集型、侵入性的并且对儿童不够敏感， 由于免疫系统不成熟，幼儿容易罹患少杆菌疾病。 结核病进展迅速，发病率和死亡率显着，评估其治疗反应。 依赖主观措施，限制了儿童获得急需的更短、更安全的结核病治疗方案。 因此，儿童迫切需要快速、严格的结核病诊断和治疗监测方法。 为了解决目前儿科结核病管理的局限性，我们建议开发一种快速诊断方法 独立于分枝杆菌分离的方法，可定量低分子量结核分枝杆菌 (Mtb) 抗原（CFP-10 和 ESAT-6）以及患者血液样本中的结核病相关宿主标记物 IP-10。 和 CFP-10 是检测活动性结核病及其治疗反应的理想生物标志物，并且将 IP-10 添加到其中 Mtb 特异性蛋白质组模式将提供患者在治疗过程中病理生理变化的综合概况。 抗结核治疗，同时还提高诊断敏感性。我们的策略利用空心、能量集中的方法。 TiO2 NanoShells 具有针对 Mtb CFP-10 和 ESAT-6 以及宿主 IP-10 的定制抗体功能 通过高通量质谱 (MS) 分析肽，以提高诊断灵敏度和特异性。 使用 292 名成人评估了该 NanoShell-MS 平台的仅 Mtb 抗原版本（无抗 IP-10 NanoShell） 以及从五个高度相关的队列（成人、儿童和潜伏性结核病、 HIV/TB 合并感染和非结核分枝杆菌感染），来自我们的全球合作者。 活动性结核病（细菌学确诊和临床诊断）的敏感性和特异性分别为： 成人为 90.7% / 97.7%，儿童为 88.2% / 100%，培养阴性儿童结核病的敏感性为 78.6% 患者，大大超过了 WHO 公布的该队列中 4-15% 的 Xpert 敏感性。 可以在样品采集后一小时内获得，而传统培养需要 4-6 周，并且 在该提案中，在开始抗结核治疗后四天内检测到血液 Mtb 抗原水平下降。 我们的目标是 1) 使用 NanoShell-MS 分析开发活动性结核病诊断的定量预测模型 大型、详细描述的前瞻性儿科结核病队列，2) 确定我们的方法在快速诊断方面的效用 我们的 NanoShell-MS 检测平台的优点是使用准确、 高通量质谱技术已成为许多地区临床诊断的重要手段 世界，同时将我们的 NanoShell-MS 平台与易于使用的“小型化”（鞋盒大小）集成 MS 系统也应该允许这种方法为资源有限地区的儿科患者提供服务。