Protein Biomarkers for Invasive Aspergillosis Diagnostics

用于侵袭性曲霉病诊断的蛋白质生物标志物

• 批准号: 8434808
• 负责人: Marta L. Feldmesser
• 金额: $ 46.26万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-01-01 至 2015-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8434808
• 关键词: Adrenal Cortex Hormones; Affect; Allergic; Allergic Bronchopulmonary Aspergillosis; Allergic Disease; Animal Model; Antibody Formation; Antifungal Agents; Antigens; Arizona; Aspergillosis; Aspergillus fumigatus; Asthma; Award; Biochemistry; Biological Assay; Biological Markers; Biophysics; Carbohydrates; Clinical; Collaborations; Communicable Diseases; Contracts; Detection; Development; Diagnosis; Diagnostic; Diagnostic Procedure; Diagnostic tests; Disease; Disease Marker; Early Diagnosis; Enzyme-Linked Immunosorbent Assay; Fever; Fungal Antibodies; Fungal Proteins; Growth; Head; Hypersensitivity; Immunoassay; In Vitro; Individual; Infection; Link; Lung diseases; Mass Spectrum Analysis; Mediating; Medicine; Methods; Modeling; Molecular; Monoclonal Antibodies; Mus; Organism; Outcome; Pathogenesis; Patients; Process; Proteins; Proteome; Proteomics; Reagent; Recombinant Proteins; Resources; Serum; Specimen; Syndrome; United States National Institutes of Health; Universities; assay development; base; biodefense; burden of illness; clinical application; college; disease diagnosis; flexibility; immunosuppressed; improved; innovation; mortality; novel diagnostics; point of care; prototype; response

ABSTRACT With the rise in susceptible individuals, the burden of disease due to Aspergillus fumigatus has increased substantially. In addition, because of the difficulties in diagnosis, the actual number of affected people likely has been severely underestimated. Improved outcome in patients with invasive aspergillosis has been directly linked to earlier diagnosis, underscoring the need for more accurate diagnostic tests. Among the challenges posed for diagnostic assays include: i. the ubiquity of the organism, necessitating the ability to distinguish between contamination, infection and disease; ii. the variety of susceptible hosts, in whom the pathogenesis of disease may vary; iii. the spectrum of disease syndromes, which range from allergy- mediated processes to aspergilloma to invasive disease. At present, the majority of effort in diagnostic development has focused on serum detection of carbohydrate antigens, PCR-based indicators of fungal burden or immunoassays for detection of anti-fungal antibody responses. Fungal proteomic analyses generally have been limited to in vitro growth conditions. This application proposes a collaboration between Dr. Marta Feldmesser (Albert Einstein College of Medicine) with Drs. John Galgiani and Vicki Wysocki (University of Arizona) to identify specific protein biomarkers associated with invasive disease in animal models of neutropenic and corticosteroid-treated mice, but that are not found in a model of allergic bronchopulmonary aspergillosis. The fungal proteome will be probed for the range of markers that will allow for more sensitive and specific markers of disease using mass spectrometry approaches to biomarker identification. Recombinant proteins then will be used to create monoclonal antibodies that can be used for mass spectrometry-based assay development. Six specific Aims are proposed: 1. To identify A. fumigatus proteins present specifically during invasive pulmonary disease and not during fungal asthma; 2. To express and purify recombinant proteins of three candidate biomarkers of invasive aspergillosis; 3. To make monoclonal antibodies to fungal proteins identified in Aim 1 for use in diagnostic assays; 4. To use mass spectrometry to identify monoclonal antibodies for use in immunoassays; 5. To develop a prototype ELISA to detect candidate biomarkers in experimental invasive and allergic aspergillosis murine models and in clinical specimens from other NIH contract resources; and 6. To begin to migrate the recombinant protein and MAb reagents to multiplex and point-of-care platforms for clinical applications. The first two Aims would be accomplished during the R21 portion of the award, while the remaining four would be done during the R33 period. The accomplishment of these studies would provide novel diagnostic methods with potential for flexibility in response to particular clinical settings.

摘要 随着易感个体的增加，烟曲霉引起的疾病负担已经增加， 大幅增加。此外，由于诊断困难，受影响的实际人数 人们可能被严重低估了。侵袭性曲霉病患者的预后改善， 与早期诊断直接相关，强调需要更准确的诊断测试。中 对诊断测定提出的挑战包括：i.生物体的普遍存在，需要有能力 区分污染、感染和疾病; ii.各种易感宿主，其中 疾病的发病机理可能不同; iii.疾病综合征的范围，从过敏- 介导的过程曲霉菌瘤的侵袭性疾病。目前，大部分的努力，在诊断 发展的重点是检测血清中的糖类抗原，基于PCR的真菌指标 用于检测抗真菌抗体应答的负荷或免疫测定。真菌蛋白质组学分析 通常限于体外生长条件。该应用程序提出了一个合作， 博士玛尔塔费尔德梅瑟（阿尔伯特爱因斯坦医学院）与博士。 （亚利桑那大学）在动物模型中鉴定与侵袭性疾病相关的特定蛋白质生物标志物 但在过敏性支气管肺炎模型中未发现 曲霉病真菌蛋白质组将被探测的范围内的标志物，将允许更敏感的 以及使用质谱法鉴定疾病的特异性标志物。 然后，重组蛋白将被用于制造单克隆抗体， 基于光谱法的测定开发。提出了六个具体目标：1。鉴定A.烟曲霉蛋白质 在侵袭性肺部疾病期间特异性地存在，而在真菌性哮喘期间不存在; 2.表达和 纯化侵袭性曲霉病三个候选生物标志物的重组蛋白; 3.为了使单克隆 用于诊断测定的针对目的1中鉴定的真菌蛋白的抗体; 4.用质谱分析法 鉴定用于免疫测定的单克隆抗体; 5.开发一种ELISA原型， 实验侵袭性和过敏性曲霉菌病小鼠模型和来自 其他NIH合同资源;和6.开始迁移重组蛋白和单克隆抗体试剂， 用于临床应用的多路复用和即时护理平台。前两个目标将在 R21部分的奖励，而其余四个将在R33期间完成。的 这些研究的完成将提供新的诊断方法，具有灵活性， 对特定临床环境的反应。