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）：这项工作的目标是开发慢性移植物抗宿主病（cGVHD）的唾液生物标志物。这将使用基于 iTRAQ 标记的定量质谱 (MS) 方法来检测接受造血细胞移植 (HCT) 并随后发生 cGVHD 的患者的静息全唾液样本中与 cGVHD 相关的蛋白表达变化。提议将唾液用于本研究，不仅是因为其获取方便，而且主要是因为唾液腺是粘膜免疫系统中淋巴细胞归巢的靶标，我们假设 GVHD 被激活（Murai 等人，Nat Immunol 4:154-160, 2003）。因此，唾液蛋白质组成的变化应该发生在疾病早期，并且应该与疾病强度相关。 GVHD 的病因预测唾液炎症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，基于 iTRAQ 的定量 MALDI TOF/TOF 质谱技术将用于在来自[有或没有 cGVHD 的 HCT 受者]的静息全唾液混合样本中识别 cGVHD 的潜在候选细胞因子/趋化因子和其他蛋白质生物标志物。在验证阶段，八种候选生物标志物蛋白显示出最大的 cGVHD 相关表达 将使用 [ELISA] 测定来测量差异，以确认合并样本中的 MS 结果，并确定个体患者样本中的表达差异幅度和患病率。个体唾液样本中的蛋白质表达变化将用于构建由 5 种蛋白质组成的生物标志物组，将评估其预测 cGVHD 的能力。 MS 分析将由我们的合作者、华盛顿大学 Shaw 神经病理学教授张静博士指导，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。我们建议应用他的技术来寻找 cGVHD 的唾液生物标志物。我们的初步结果表明对照全唾液可以通过建议的 MS 程序进行分析。所提出的最先进的 MS 蛋白质组学方法应该产生特定的 GVHD 生物标志物，从而可以更及时地检测 GVHD，从而实现早期治疗，从而减轻口腔和其他 cGVHD 靶组织的疾病严重程度。 [由于许多疾病生物标志物源自潜在的病理生理机制，因此对此处发现的一系列生物标志物进行功能分析应该可以深入了解 cGVHD 的发病机制。] 移植物抗宿主病 (GVHD) 是用于治疗血液系统疾病和其他疾病的造血细胞移植的一种常见、严重、可能危及生命的并发症。 GVHD 可以作为急性或慢性疾病发生，目前很难预测谁会发展为广泛的疾病。拟议的疾病病因表明唾液腺应该早期参与 GVHD，唾液蛋白的变化可以实现早期检测和治疗，从而降低疾病的严重性和死亡率，[并深入了解疾病病因]。该提案的目标是开发这样的唾液检测方法。