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: 8583121
• 负责人: Susan E Dorman
• 金额: $ 26.32万
• 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 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8583121
• 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。尽管已经进行了基于痰液的TB诊断测试的增量进展，但在临床上易于访问的标本（如血液和尿液）中，具有高灵敏度的结核分枝杆菌（MTB）抗原的能力将改变TB诊断过程。 MTB脂质是有吸引力的诊断靶标，虽然有几条证据表明，在结核病患者中，血液和尿液中存在某些MTB脂质，但生物学仍未开发，并且诊断潜力未实现。我们的团队开发了一个基于波导的光学生物传感器平台，用于敏感的MTB脂肪脂动脉瘤（LAM）。此外，我们基于其两亲性质，并与受支持的脂质双层的相互作用开发了一种新型的血清中MTB LAM的超敏感检测策略。少量临床标本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（尤其是高密度脂蛋白（HDL））包装在脂质组件中，从而增加了人脂蛋白作为AmphipHilic MTB产品的“下沉”的可能性。通过利用两亲性LAM的这种相互作用，我们制定了一种从血清中选择性下拉的策略。这种新颖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）完善我们在临床标本中使用现有的血清和尿液标本中发现LAM的方法的方法，并从具有良好特征的TB疾病状态的成年人中，b）对LAM与人类脂肪蛋白的相互作用进行体外观察，以扩展LAM与其他LAM的生物作用，并确定LAM与人类的生物作用。拟议的研究不仅可以通过发现MTB脂质的新型超敏化方法来取得突破，而且在结核病和人类生物学方面，由于人类血浆脂蛋白在分枝杆菌脂质贩运中的作用，因此无法探索。拟议的探索性研究有望为开发快速诊断测试平台的发展奠定基础，以及旨在阐明LAM和其他分枝杆菌病原体相关的分子模式的生物学和治疗意义的未来全面的，基于假设的研究。现有的研究临床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的现有临床标本增强了该项目的可行性。该项目直接源于NIAID/DMID资助的结核病临床诊断研究联盟合同中研究人员的技术寻求活动。