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: 10372903
• 负责人: Christopher J Lyon
• 金额: $ 38.68万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-09 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10372903
• 关键词: Address; Adult; Antibodies; Antigens; Antitubercular Agents; Bacillus; Bacteria; Bacteriology; Biological Assay; Biological Markers; Blood; Blood Volume; Blood specimen; CXCL10 gene; Chest; Child; Childhood; Clinical; Clinical/Radiologic; Consumption; Custom; DNA; Data; Diagnosis; Diagnostic; Diagnostic Procedure; Diagnostic Sensitivity; Disease; Disease Progression; Early Diagnosis; Enrollment; Evaluation; Exposure to; Extreme drug resistant tuberculosis; Growth; HIV; HIV/TB; Health; Hour; Immune response; Immunologics; Infection; Investigation; Longitudinal Studies; Mass Spectrum Analysis; Measures; Methods; Microscopy; Molecular Weight; Monitor; Morbidity - disease rate; Mycobacterium Infections; Mycobacterium tuberculosis; Mycobacterium tuberculosis antigens; National Institute of Child Health and Human Development; Noise; Organism; Patients; Pattern; Peptide Signal Sequences; Peptides; Performance; Physicians; Population; Prospective cohort; Proteins; Proteomics; Publishing; Rapid diagnostics; Reaction Time; Regimen; Research Infrastructure; Resource-limited setting; 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; Tuberculosis diagnosis; Validation; Work; accurate diagnosis; base; biobank; biomarker signature; clinical diagnosis; clinically relevant; co-infection; cohort; detection method; detection sensitivity; diagnostic value; early detection biomarkers; high risk; improved; improved outcome; interest; miniaturize; mortality; mycobacterial; nanoparticle; nanoshell; non-tuberculosis mycobacteria; novel; pediatric patients; predict clinical outcome; predictive modeling; profiles in patients; prospective; proteomic signature; recruit; response; sample collection; side effect; titanium dioxide; treatment comparison; 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 成人和小儿结核病（TB）诊断和治疗监测的常规方法严重依赖 在耗时的细菌培养或不可量化的DNA分析上，以检测少量的存在 细菌。小儿结核病管理特别困难，因为当前基于生物的方法用于 诊断和治疗监测对儿童的资源密集，侵入性和不充分敏感， 谁倾向于患有paucibaCillarry疾病。由于免疫学不成熟，幼儿有高风险 快速疾病结核病进展，发病率和死亡率很高。评估他们的治疗反应 依靠主观措施，限制儿童获得急需的较短和更安全的结核病方案。更多的 因此，儿童需要快速，严格的结核病诊断和治疗监测方法。 为了解决小儿结核病管理中当前限制，我们建议开发快速诊断 与分枝杆菌分离无关的方法，可以量化低分子量的结核分子（MTB） 患者血液样本中的抗原（CFP-10和ESAT-6）以及与TB相关的宿主标记IP-10。 ESAT-6 CFP-10是检测活性结核病及其治疗反应的理想生物标志物，并在此增加IP-10 MTB特异性的蛋白质组学模式将提供患者病理生理变化期间的综合概况 抗TB治疗，同时也提高了诊断敏感性。我们的策略利用空心，以能源为目标 TIO2纳米壳，可与MTB CFP-10和ESAT-6和主机IP-10一起功能化的定制抗体 具有高通量质谱（MS）的肽，以提高诊断敏感性和特异性。我们 与292个成人的Nanoshell-MS平台（无抗IP-10纳米壳）评估了仅MTB抗原的版本（无抗IP-10纳米壳） 从五个高度相关的队列中选择的102名儿科患者和对照组（成人，小儿和潜在结核， 来自我们的全球合作者，HIV/TB共感染和非TB分枝杆菌感染）。 Nanoshell-MS 主动结核的敏感性和特异性（细菌学确认和临床诊断）分别为 成人的90.7％ / 97.7％，儿童为88.2％ / 100％，培养阴性小儿TB的敏感性为78.6％ 在此队列中，患者大大超过了WHO出版的4-15％XPERT敏感性。 Nanoshell-MS结果 可以在采样后一小时内获得，而常规培养物的4-6周则可以获得 在抗TB治疗计划的四天内，血液MTB抗原水平的下降发现。在此提案中 我们的目的是1）使用纳米壳-MS分析开发一个定量预测模型，以进行主动结核病诊断 大型，描述良好的前瞻性小儿TB队列和2）确定我们方法的实用性 评估治疗效率。我们的Nanoshell-MS分析平台的优势是它使用准确的， 高通量质谱法，这已成为许多部分临床诊断的关键手段 世界，虽然我们的纳米壳-MS平台与易于使用的“小型化”（鞋盒尺寸）集成 MS系统还应允许这种方法在资源有限的地区为儿科患者提供服务。