Host Proteomic Biosignatures for a Urine-based Diagnosis of Pulmonary Tuberculosis in Children

项目摘要 结核病（TB）是全世界儿童死亡的主要原因。痰液采集困难， 痰菌载量是诊断的主要障碍，并且需要发展非痰菌， 儿童胸内结核病的生物标志物检测儿童结核病的宿主生物标志物发现 需要更多地关注下游蛋白质及其翻译后修饰（PTM）， 可能对疾病表型具有特异性，并且可以更容易地转化为即时检测。与 为了支持这项K23奖，Devan Jaganath博士将鉴定尿液中的宿主蛋白质组生物特征， 实现儿童肺结核分诊和/或诊断测试的目标准确性。完成这个 目的，他将利用一个正在进行的前瞻性队列的症状儿童正在评估， 乌干达坎帕拉的胸内结核病研究， 加州，旧金山弗朗西斯科（加州大学旧金山分校）。在目标1中，他将对来自 儿童确诊与不太可能结核病，并检查10个宿主蛋白质的丰度和泛素化， 有与M有特定相互作用的先前证据。结核病（Mtb）蛋白作为候选生物标志物。在Aim中 2，他将使用鸟枪质谱对尿液样本，以确定所有的宿主蛋白质和他们的PTM， 可以区分TB状态作为新的生物标志物，并进行途径分析，以确定子集， 与Mtb发病机制的功能相关性。在目标3中，他将应用机器学习分析来识别 可以实现用于分诊和/或诊断的目标准确度阈值的生物标志物的最小组合 检测胸腔内结核病然后，他将评估有希望的生物签名在一个 独立、前瞻性入组的测试集。通过这种方法，Jaganath博士寻求优化生物标志物 通过将前瞻性临床队列与靶向和非靶向的 高通量方法，以全面检查非痰样本的宿主生物标志物的儿童。 博士Jaganath的职业目标是成为一名医生科学家，将非痰生物标志物转化为临床工具 可以改善对结核病儿童的治疗。为了支持他的独立之路，拟议的工作将是 与专门的多学科指导团队和国际儿科结核病生物标志物培训相结合 研究，蛋白质组学分析的生物信息学和机器学习。UCSF是一个出色的环境， 致力于初级研究人员的研究和职业发展的广泛资源，和Mulago 乌干达国家转诊医院是儿科结核病研究的领导者， 用于正在进行的招募和样本采集。研究结果将支持NIH R 01应用程序，以验证 与现有的非痰诊断相比，在大的、不同的群组中的生物标志物和生物特征。因此，在本发明中， K23奖将为Jaganath博士提供重要的指导、培训、资源和经验， 成为一名独立的研究人员，可以为儿童结核病领域做出重要贡献。

PROJECT ABSTRACT Tuberculosis (TB) is a leading cause of mortality in children worldwide. Difficulty in obtaining sputum and a low sputum bacillary load are major barriers to diagnosis, and necessitate the development of a non-sputum, biomarker-based assay for childhood intrathoracic TB disease. Host biomarker discovery for childhood TB requires a greater focus on downstream proteins and their post-translational modifications (PTMs) that are more likely to be specific to a disease phenotype and can be more easily translated into a point-of-care test. With the support of this K23 award, Dr. Devan Jaganath will identify a host proteomic biosignature in urine that can achieve the goal accuracy for a triage and/or diagnostic test for pulmonary TB in children. To complete this objective, he will leverage an ongoing prospective cohort of symptomatic children being evaluated for intrathoracic TB in Kampala, Uganda, with the extensive proteomic facilities available at the University of California, San Francisco (UCSF). In Aim 1, he will perform targeted mass spectrometry on urine samples from children with confirmed vs. unlikely TB, and examine the abundance and ubiquitylation of 10 host proteins that have prior evidence of specific interactions with M. tuberculosis (Mtb) proteins as candidate biomarkers. In Aim 2, he will use shotgun mass spectrometry on the urine samples to identify all host proteins and their PTMs that can differentiate TB status as novel biomarkers, and perform pathway analysis to determine the subset with functional relevance to Mtb pathogenesis. In Aim 3, he will apply machine learning analyses to identify the smallest combination of biomarkers that can achieve the target accuracy thresholds for a triage and/or diagnostic test for intrathoracic TB disease. He will then evaluate the performance of promising biosignatures in an independent, prospectively enrolled test set. Through this approach, Dr. Jaganath seeks to optimize biomarker discovery for childhood TB diagnosis by coupling prospective clinical cohorts with a targeted and untargeted high-throughput approach to comprehensively examine non-sputum samples for host biomarkers for children. Dr. Jaganath's career goal is to be a physician scientist who translates non-sputum biomarkers into clinical tools that can improve the care of children with TB. To support his path to independence, the proposed work will be paired with a dedicated, multidisciplinary mentorship team and training in international pediatric TB biomarker studies, bioinformatics for proteomic analysis, and machine learning. UCSF is an outstanding environment that is committed to junior investigators with extensive resources for research and career development, and Mulago National Referral Hospital in Uganda is a leader in pediatric TB research, and has the established infrastructure for ongoing enrollment and sample collection. The findings will support an NIH R01 application to validate the biomarkers and biosignatures in large, diverse cohorts in comparison to existing non-sputum diagnostics. Thus, the K23 award will provide Dr. Jaganath with the critical mentorship, training, resources and experience to become an independent investigator who can make important contributions to the field of childhood TB.