Detecting pathogen and host factors on extracellular vesicles for pediatric TB diagnosis and management

检测细胞外囊泡上的病原体和宿主因子，用于儿童结核病的诊断和管理

• 批准号: 10753281
• 负责人: Tony Y. Hu
• 金额: $ 63.24万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 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 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10753281
• 关键词: Adult; Aftercare; Antitubercular Agents; Award; Bacillus; Biological; Biological Assay; Biological Markers; Blood specimen; Body Fluids; Breakthrough device; Cells; Cessation of life; Characteristics; Child; Childhood; Chronic Disease; Collaborations; Complex; Containment; DNA; Data; Detection; Development; Devices; Diagnosis; Diagnostic; Disease; Dominican Republic; Early Diagnosis; Enrollment; Equipment; Evaluation; Exhibits; Guidelines; HIV; HIV Infections; Hospitals; Immune; Immune System Diseases; Immune response; Immune system; Immunoassay; Individual; Infant; Integration Host Factors; International Maternal Pediatric Adolescent AIDS Clinical Trials; Lung; Macrophage; Mass Spectrum Analysis; Measurement; Methods; Modification; Monitor; Morbidity - disease rate; Mycobacterium tuberculosis; Outcome; Pathogen detection; Pathogenesis; Pattern; Pediatric cohort; Performance; Phagocytes; Play; Population; Procedures; Process; Prospective cohort; Proteins; Proteomics; Reporting; Reproducibility; Resource-limited setting; Retrospective cohort; Risk; Role; Sampling; Serum; Signal Transduction; Sputum; Surface; Testing; Time; Translations; Tuberculosis; Tuberculosis diagnosis; Universities; Validation; Variant; biomarker signature; blood-based biomarker; clinical application; cohort; effectiveness evaluation; efficacy evaluation; experience; extracellular vesicles; improved; lifetime risk; machine learning algorithm; mass spectrometer; mortality; mycobacterial; nanoparticle; pathogen; portability; predictive modeling; preservation; prevent; prognostic performance; prospective; respiratory; treatment response; tuberculosis treatment; validation studies

ABSTRACT Tuberculosis (TB) is a major cause of pediatric morbidity and mortality (1.2 million new cases and 224,000 deaths in 2021), with most deaths occurring in children < 5 years who do not initiate treatment. These children often present with disseminated or extrapulmonary TB as their immune systems may fail to contain their M. tuberculosis (Mtb) infections in their lungs. Since host responses play critical roles in TB development, analysis of Mtb- and host-derived could improve early diagnosis and TB management. However, TB tests still primarily rely on microbiologic evidence from respiratory samples that are difficult to obtain from, and less informative for, young children. Sputum culture detects only 30-60% of pediatric TB, and similar results are obtained with PCR- based Xpert MTB/RIF and Xpert Ultra assays. There is thus an urgent need for non-sputum-based tests that can effectively diagnose pediatric TB and monitor its response to treatment. Serum biomarker assays are ideal for pediatric TB diagnosis. The assay used in our initial proposal detects low abundance Mtb proteins in serum to effectively diagnose pediatric TB and has since received breakthrough device status by the FDA prior to pending validation studies for its FDA clearance. We have further shown that we can employ a nanoparticle-enhanced immunoassay (NEI) read by an inexpensive portable device to detect Mtb-derived factors on extracellular vesicles (EVs) for TB diagnosis, an approach suitable for use in resource- limited settings. We have also shown that a simple modification can markedly increase sensitivity and reproducibility to improve measurements. Biomarkers on EVs secreted by Mtb-infected phagocytes and by immune cells may reflect Mtb burden and its containment by the immune system, and provide critical information to required improve pediatric TB diagnosis and to allow rapid and accurate assessment of effective anti-TB treatment. However, few studies have evaluated the diagnostic and prognostic performance EV-based Mtb assays, and none have evaluated the ability of host-derived EV biomarkers to improve assay performance. We now propose modify our initial NEI test to incorporate both Mtb- and host-derived EV markers associated with TB disease in order to enhance its performance for TB diagnosis and treatment monitoring. We will employ the following aims to develop and characterize this modified assay: 1) We will optimize its EV biomarker signature using additional Mtb- and host-derived proteins we will identify from serum EVs of TB cases and controls. 2) We will conduct an analytical validation of this refined NEI to establish its characteristics using STARD guidelines. 3) We will establish a prediction model of NEI array’s multiplex performance for TB diagnosis and treatment evaluation using longitudinal serum and data from a retrospective cohort of HIV-exposed infants at increased risk for TB (IMPAACT P1041 cohort) with longitudinal samples. 4) We will validate the NEI array’s ability to diagnose LTBI and active TB and evaluate the efficacy of anti-TB treatment responses using longitudinal serum from a second cohort of older children prospectively enrolled at the O&M Hospital in the Dominican Republic.

摘要 结核病（TB）是儿科发病率和死亡率的主要原因（120万新发病例和224，000 2021年死亡），大多数死亡发生在未开始治疗的5岁以下儿童中。这些孩子 由于他们的免疫系统可能无法控制他们的M。 肺结核（Mtb）感染。由于宿主反应在结核病发展中起着关键作用， Mtb和宿主来源的结核分枝杆菌的检测可以改善结核病的早期诊断和管理。然而，结核病检测仍然主要 依赖于呼吸道样本的微生物证据，这些样本很难获得，而且信息量较少， 年幼的孩子。痰培养只能检出30-60%的儿童结核病，PCR也能得到类似的结果。 基于Xpert MTB/RIF和Xpert Ultra检测试剂盒。因此，迫切需要非痰基测试， 有效诊断儿童结核病并监测其对治疗的反应。 血清生物标志物检测是儿科结核病诊断的理想方法。在我们最初的建议中使用的检测方法检测 血清中低丰度结核分枝杆菌蛋白，以有效地诊断儿科结核病，并已获得突破 在等待FDA批准的验证研究之前，FDA对器械状态进行了评估。我们进一步表明， 我们可以使用一种廉价的便携式设备读取纳米颗粒增强免疫测定（NEI）， 细胞外囊泡（EV）上的结核杆菌衍生因子用于结核病诊断，这是一种适合用于资源的方法- 有限的设置。我们还表明，一个简单的修改可以显着提高灵敏度， 重现性，以改善测量。由Mtb感染的吞噬细胞分泌的EV上的生物标志物和由Mtb感染的吞噬细胞分泌的EV上的生物标志物 免疫细胞可以反映Mtb的负荷和免疫系统对其的遏制，并提供关键信息 需要改进儿科结核病诊断，并允许快速准确地评估有效的抗结核药物， 治疗然而，很少有研究评估基于EV的Mtb的诊断和预后性能， 然而，没有人评估宿主来源的EV生物标志物改善测定性能的能力。 我们现在建议修改我们最初的NEI测试，以纳入结核分枝杆菌和宿主衍生的EV标志物， 以提高其在结核病诊断和治疗监测方面的性能。我们会委聘 以下目的是开发和表征这种改进的测定法：1）我们将优化其EV生物标志物特征 使用另外的Mtb和宿主衍生蛋白，我们将从TB病例和对照的血清EV中鉴定。2)我们 我们将对这一改进的NEI进行分析验证，以使用STARD指南确定其特性。 3)我们将建立NEI阵列在结核病诊断和治疗中的多重性能预测模型 使用纵向血清和来自一个回顾性队列的HIV暴露婴儿的数据进行评估， TB风险（IMPAACT P1041队列），纵向样本。4)我们将验证NEI阵列的能力， 诊断LTBI和活动性TB，并使用纵向血清评估抗TB治疗反应的疗效 来自多米尼加共和国O&M医院前瞻性招募的第二组年龄较大的儿童。

项目成果

Rapid Lipid-Based Approach for Normalization of Quantum-Dot-Detected Biomarker Expression on Extracellular Vesicles in Complex Biological Samples.

• DOI: 10.1021/acs.nanolett.9b02232
• 发表时间: 2019-11-13
• 期刊: Nano letters
• 影响因子: 10.8
• 作者: Rodrigues M;Richards N;Ning B;Lyon CJ;Hu TY
• 通讯作者: Hu TY

Strategies for advanced personalized tuberculosis diagnosis: Current technologies and clinical approaches.

• DOI: 10.1093/pcmedi/pbaa041
• 发表时间: 2021-03
• 期刊: Precision clinical medicine
• 影响因子: 5.3
• 作者: Chen X;Hu TY
• 通讯作者: Hu TY

From Prevention to Therapy: A Roadmap of Nanotechnologies to Stay Ahead of Future Pandemics.

• DOI: 10.1021/acsnano.2c04148
• 发表时间: 2022-07-26
• 期刊: ACS NANO
• 影响因子: 17.1
• 作者: Chandra, Sutapa;Hu, Tony
• 通讯作者: Hu, Tony

Mesoporous silica chip: enabled peptide profiling as an effective platform for controlling bio-sample quality and optimizing handling procedure.

介孔二氧化硅芯片：使肽分析成为控制生物样品质量和优化处理程序的有效平台。

• DOI: 10.1186/s12014-016-9134-9
• 发表时间: 2016
• 期刊: Clinical proteomics
• 影响因子: 3.8
• 作者: Liang,Kai;Wu,Hongmei;Hu,TonyY;Li,Yan
• 通讯作者: Li,Yan

Predictive value of serum bradykinin and desArg9-bradykinin levels for chemotherapeutic responses in active tuberculosis patients: A retrospective case series.

• DOI: 10.1016/j.tube.2016.09.022
• 发表时间: 2016-12
• 期刊: Tuberculosis
• 影响因子: 3.2
• 作者: X. Qian;D. Nguyen;Yaojun Li;J. Lyu;E. Graviss;Tony Y. Hu