Salivary Biomarkers for Graft versus Host Disease

移植物抗宿主病的唾液生物标志物

• 批准号: 7383733
• 负责人: Kenneth T Izutsu
• 金额: $ 23.4万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-03-01 至 2010-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7383733
• 关键词: Activated Lymphocyte; Acute; Arts; Ascaridil; Biological Assay; Biological Markers; Cell Transplants; Cells; Central Nervous System Diseases; Chemokine, Other; Chronic Disease; Class; Clinical; Complication; Computer software; Detection; Diagnosis; Disease; Early Diagnosis; Early treatment; Enzyme-Linked Immunosorbent Assay; Etiology; Functional disorder; Goals; Guidelines; Hematopoietic; Hematopoietic stem cells; Immune Cell Activation; Immune system; Individual; Inflammatory; Injury; Laboratories; Lead; Life; Lymphocyte; Mass Spectrum Analysis; Measurement; Measures; Methods; Morbidity - disease rate; Oral; Pathogenesis; Pathway interactions; Patients; Phase; Prevalence; Procedures; Proteins; Proteomics; Recording of previous events; Research; Rest; Saliva; Salivary; Salivary Glands; Salivary Proteins; Sampling; Severity of illness; Signal Transduction; Signaling Protein; Specificity; Spectrometry; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Staging; Stem cell transplant; Structure of aggregated lymphoid follicle of small intestine; System; Techniques; Testing; Tissues; Transplant Recipients; United States National Institutes of Health; Validation; Widespread Disease; Work; base; chemokine; chronic graft versus host disease; cytokine; graft vs host disease; insight; mortality; neuropathology; polypeptide; professor; protein expression; protein function; response; salivary assay

DESCRIPTION (provided by applicant): The goal of this work is to develop salivary biomarkers for chronic Graft Versus Host Disease (cGVHD). This will be done using an iTRAQ marker-based, quantitative mass spectrometry (MS) approach to detect cGVHD-associated protein expression changes in resting whole saliva samples from patients who received hematopoietic cell transplants (HCT) and subsequently developed cGVHD. Saliva is proposed for this study not simply because it is convenient to obtain, but primarily because salivary glands are a lymphocyte-homing target in the mucosal immune system, where we postulate GVHD is activated (Murai et al., Nat Immunol 4:154-160, 2003). Hence, salivary protein composition changes should occur early in the disease and should correlate with disease intensity. The etiology of GVHD predicts increases in salivary inflammatory cytokine and related cell signaling protein contents. Our study will consist of discovery and confirmation/validation phases. In the discovery phase, an iTRAQ-based quantitative MALDI TOF/TOF mass spectrometry technique will be used to identify potential candidate cytokine/chemokine and other protein biomarkers for cGVHD in pooled samples of resting whole saliva from [HCT recipients with or without cGVHD.] In the validation phase, the eight candidate biomarker proteins showing the greatest cGVHD-associated expression differences will be measured using [ELISA] assays to confirm the MS results in the pooled samples, and to establish expression difference magnitudes and prevalence in the individual patient samples. The protein expression changes in the individual saliva samples will be used to construct a biomarker panel of 5 proteins that will be evaluated for an ability to predict cGVHD. The MS analyses will be directed by our collaborator, Dr. Jing Zhang, Shaw Professor of Neuropathology at the UW, who has developed MS techniques for identifying CSF biomarkers for central nervous system diseases. We propose to apply his techniques to find salivary biomarkers for cGVHD. We have preliminary results showing control whole saliva can be analyzed by the proposed MS procedures. The proposed state-of-art MS proteomics approach should yield specific GVHD biomarkers that would allow more timely detection of GVHD, leading to earlier treatment, and consequently less disease severity in the oral and other target tissues of cGVHD. [Since many disease biomarkers arise from underlying pathophysiologic mechanisms, function-analysis of the array of biomarkers found here should give insight into the pathogenesis of cGVHD.] Graft Versus Host Disease (GVHD) is a frequent, serious, potentially life-threatening complication of hematopoietic cell transplants used to treat hematologic disorders as well as other diseases. GVHD can occur as acute or chronic disease, and it is currently difficult to predict who will develop extensive disease. The proposed etiology of the disease suggests salivary glands should have an early involvement in GVHD, and salivary protein changes could allow early detection and treatment, which could decrease the seriousness and mortality of the disease, [as well as give insight into disease etiology]. The goal of this proposal is to develop such a salivary assay.

描述（由申请人提供）：这项工作的目的是为慢性移植与宿主疾病（CGVHD）开发唾液生物标志物。这将使用基于ITRAQ标记的定量质谱法（MS）方法来检测来自接受造血细胞移植（HCT）的患者的静息整个唾液样品的蛋白质表达变化并随后开发CGVHD。为这项研究提出了唾液，这不仅是因为它很方便，而且主要是因为唾液腺是粘膜免疫系统中的淋巴细胞 - 含量靶标，我们假设GVHD被激活（Murai等，Nat Immunol 4：154-160，2003）。因此，唾液蛋白成分的变化应在疾病的早期发生，并应与疾病强度相关。 GVHD的病因可以预测唾液炎性细胞因子和相关细胞信号蛋白含量的增加。我们的研究将包括发现和确认/验证阶段。在发现阶段，将使用基于ITRAQ的定量MALDI TOF/TOF/TOF质谱技术来鉴定潜在的候选候选细胞因子/趋化因子和其他蛋白质生物标志物，用于CGVHD，用于在均衡的静脉内唾液中汇集的整个唾液样本中的CGVHD [无与伦比的HCT接受者中均无CGVHD的HCT。将使用[ELISA]测定法确认汇总样品中的MS结果，并​​在单个患者样本中确定表达差的幅度和患病率，从而测量差异。单个唾液样品中的蛋白质表达变化将用于构建5种蛋白质的生物标志物面板，以预测CGVHD的能力。 MS分析将由我们的合作者，UW神经病理学教授Jing Zhang博士指导，他开发了用于识别中枢神经系统疾病的CSF生物标志物的MS技术。我们建议采用他的技术来寻找CGVHD的唾液生物标志物。我们的初步结果表明，可以通过建议的MS程序来分析整个唾液的控制。拟议的最新MS蛋白质组学方法应产生特定的GVHD生物标志物，以允许更及时检测GVHD，从而导致较早的治疗，从而在CGVHD的口腔和其他目标组织中疾病严重程度较小。 [由于许多疾病生物标志物是由潜在的病理生理机制引起的，因此此处发现的一系列生物标志物的功能分析应深入了解CGVHD的发病机理。 GVHD可以作为急性或慢性病发生，目前很难预测谁将发展广泛的疾病。该疾病的拟议病因表明，唾液腺应尽早参与GVHD，而唾液蛋白的变化可以允许早期检测和治疗，这可能会降低疾病的严重性和死亡率，并深入了解疾病病因学]。该提案的目的是开发这种唾液分析。