Lipidated Mycobacterium tuberculosis biomarkers in clinical specimens

临床标本中的脂化结核分枝杆菌生物标志物

• 批准号: 8660620
• 负责人: Susan E Dorman
• 金额: --
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-05-15 至 2016-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8660620
• 关键词: Address; Adult; Algorithms; Antibodies; Antigens; Apolipoprotein A-I; Biodistribution; Biological Assay; Biological Markers; Biology; Biosensor; Blood; Cattle; Cessation of life; Chemicals; Chemistry; Clinical; Clinical Microbiology; Collaborations; Communicable Diseases; Contracts; Detection; Development; Diagnosis; Diagnostic; Diagnostic tests; Diagnostics Research; Disease; Engineering; Enzyme-Linked Immunosorbent Assay; Funding; Future; Goals; HIV; High Density Lipoproteins; Human; Human Biology; Immune response; Immunoassay; Immunocompromised Host; Immunology; In Vitro; Institutional Review Boards; Knowledge; Label; Laboratories; Lateral; Lead; Lipid Bilayers; Lipid Chemistry; Lipids; Lipoprotein (a); Lipoproteins; Manufacturer Name; Membrane; Methods; Microscopy; Molecular; Morbidity - disease rate; Mycobacterium tuberculosis; Mycobacterium tuberculosis antigens; National Institute of Allergy and Infectious Disease; Nature; Optics; Pathogenesis; Patients; Pattern; Plasma; Process; Research Personnel; Role; Sampling; Serum; Specimen; Sputum; Technology; Testing; Therapeutic; Tuberculosis; Uganda; United States National Institutes of Health; Urine; Validation; Work; base; chemotherapy; cohort; disease diagnosis; innovation; lipid transport; lipoarabinomannan; mortality; multidisciplinary; mycobacterial; novel; novel strategies; particle; pathogen; prospective; public health relevance; rapid detection; sensor

DESCRIPTION (provided by applicant): Tuberculosis (TB) remains a leading cause of infectious disease morbidity and mortality. While incremental advances in sputum-based TB diagnostic tests have been made, the ability to rapidly detect M. tuberculosis (MTB) antigens with high sensitivity in clinically accessible specimens such as blood and urine would transform the TB diagnostic process. MTB lipids are attractive diagnostic targets, and while several lines of evidence indicate that certain MTB lipids are present in blood and urine in TB patients, the biology remains unexplored and the diagnostic potential unrealized. Our team has developed a waveguide-based optical biosensor platform for sensitive detection of MTB lipoarabinomannan (LAM). Further, we have developed a novel ultrasensitive detection strategy for monomeric MTB LAM in serum, based on its amphiphilic nature and consequent interaction with supported lipid bilayers. Preliminary work with a small number of clinical specimens is promising. Moreover, in vitro studies have shown that LAM in serum is packaged in lipid assemblies with human lipoproteins, especially high-density lipoprotein (HDL), raising the possibility that human lipoproteins serve as "sinks" for amphiphilic MTB products. By exploiting this interaction of amphiphilic LAM, we have developed a strategy for selective pull-down of LAM from serum; this novel strategy could be applied to other biomarkers. The overall goals of this R21 project are a) to refine our approach for detection of LAM in clinical specimens using existing serum and urine specimens from adults with well-characterized TB disease status, and b) to extend preliminary in vitro observations on the interaction of LAM with human lipoproteins to clarify the biodistribution of LAM and determine if other human lipoproteins are associated with LAM. The proposed studies could lead to a breakthrough not only with respect to novel ultrasensitive approaches for TB diagnosis through detection of MTB lipids, but also with respect to TB and human biology since the role of human plasma lipoproteins in mycobacterial lipid trafficking is unexplored. The proposed exploratory studies are expected to lay the groundwork for development of a rapid diagnostic testing platform as well as future comprehensive, hypothesis-based studies aimed at elucidating the biology and therapeutic implications of LAM and other mycobacterial pathogen-associated molecular patterns. The feasibility of this project is enhanced by the existing clinical specimens for study, as well as the multidisciplinary team including engineers, chemists, and TB clinicians. This project arose directly from technology-seeking activities of investigators in the NIAID/DMID-funded TB Clinical Diagnostics Research Consortium contract.

描述（由申请人提供）：结核病（TB）仍然是传染病发病率和死亡率的主要原因。虽然在结核病诊断检测方面取得了进展，但快速检测M。在临床上可获得的样本如血液和尿液中具有高灵敏度的结核病（MTB）抗原将改变TB诊断过程。MTB脂质是有吸引力的诊断靶点，虽然一些证据表明某些MTB脂质存在于TB患者的血液和尿液中，但其生物学尚未探索，诊断潜力尚未实现。我们的团队开发了一种基于波导的光学生物传感器平台，用于灵敏检测MTB脂阿拉伯甘露聚糖（LAM）。此外，我们已经开发了一种新的超灵敏的检测策略的单体MTB LAM在血清中，基于其两亲性和随之而来的相互作用与支持的脂质双层。用少量临床标本进行的初步工作是有希望的。此外，体外研究表明，血清中的LAM与人脂蛋白，特别是高密度脂蛋白（HDL）一起包装在脂质组装体中，提高了人脂蛋白作为两亲性MTB产物“汇”的可能性。通过利用两亲性LAM的这种相互作用，我们已经开发了一种从血清中选择性下拉LAM的策略;这种新的策略可以应用于其他生物标志物。该R21项目的总体目标是a）使用来自具有良好表征的TB疾病状态的成人的现有血清和尿液样本来改进我们用于检测临床样本中的LAM的方法，以及B）扩展关于LAM与人脂蛋白的相互作用的初步体外观察，以澄清LAM的生物分布并确定其他人脂蛋白是否与LAM相关。拟议的研究可能导致突破，不仅在新的超灵敏的方法，结核病诊断通过检测结核分枝杆菌脂质，而且在结核病和人类生物学方面，因为人类血浆脂蛋白在分枝杆菌脂质运输的作用是未开发的。预计拟议的探索性研究将为快速诊断检测平台的开发以及未来旨在阐明LAM和其他分枝杆菌病原体相关分子模式的生物学和治疗意义的综合性、基于假设的研究奠定基础。该项目的可行性通过现有的临床研究标本以及包括工程师、化学家和结核病临床医生在内的多学科团队得到增强。该项目直接源于NIAID/DMID资助的结核病临床诊断研究联合会合同中研究人员的技术寻求活动。