Discovering tissue-specific biomarkers of radiation injury in SILAC-labeled mice

在 SILAC 标记的小鼠中发现辐射损伤的组织特异性生物标志物

• 批准号: 8662694
• 负责人: AMANDA G PAULOVICH
• 金额: $ 44万
• 依托单位: FRED HUTCHINSON CANCER RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-06-06 至 2017-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8662694
• 关键词: Accidents; Acute; Biological Assay; Biological Markers; Blood; Blood Circulation; Blood Tests; Bone Marrow; Cause of Death; Cessation of life; Childhood; Clinical; Creatine Kinase; Development; Devices; Diagnosis; Diagnostic; Dose; Dose-Rate; Effectiveness; Elderly; Emergency Care; Emergency Situation; Ensure; Environmental Risk Factor; Extravasation; Gender; Goals; Health; Hematopoiesis; Hematopoietic; Hematopoietic System; Hospitals; Hour; Human; Immunoassay; Individual; Injury; Institution; Label; Lipase; Marrow; Measures; Medical; Methods; Modality; Mus; Muscle; Myocardial Infarction; Nuclear; Organ; Pancreas; Panic; Patients; Plasma; Population Heterogeneity; Process; Proteins; Proteomics; Radiation; Radiation Injuries; Radiation Sicknesses; Recording of previous events; Residual state; Risk; Severities; Signal Transduction; Signs and Symptoms; Specificity; Stem cell transplant; Supportive care; Syndrome; System; Techniques; Technology; Testing; Therapeutic; Time; Tissues; Transaminases; Triage; Troponin; Validation; base; care systems; cytokine therapy; injured; innovation; irradiation; liver injury; novel; point of care; prevent; response; success; urban area

DESCRIPTION (provided by applicant): At total body radiation exposures of 3-8 Gy, the predominant cause of death is the hematopoietic syndrome. Many of these deaths are preventable with rapid triage of victims for cytokine therapies and aggressive supportive care. Unfortunately current modalities for identifying patients at risk for the hematopoietic syndrome suffer from inaccuracy, high expense, long analysis times, and delayed diagnosis. Furthermore, none of these methods directly measures radiation damage to the bone marrow, nor do they indicate the existence of residual hematopoiesis, and most are not amenable to point-of-care in emergency conditions. Thus, there is a critical unmet need for a field-deployable diagnostic for use following a radiological incident to identify victims who will develop the potentially fatal ye often treatable hematopoietic syndrome. We propose to build upon a highly successful paradigm in clinical diagnostics, which is that injured tissues leak, shed, and/or secrete proteins into the plasma, where they are useful biomarkers indicating the extent of tissue injury. Precedents include the troponins in myocardial infarction, transaminases in liver injury, creatine kinase in muscle injury, and lipase in pancreatic processes. Accordingly, we hypothesize that following radiation-induced injury to the bone marrow, proteins are released from the marrow into the bloodstream, where they provide useful biomarker signals predictive of the onset and severity of the hematopoietic syndrome. We propose to identify these plasma biomarkers of radiation-induced marrow injury using an innovative approach incorporating targeted proteomic technologies that we have developed and validated in a biomarker discovery pipeline that will substantially increase our chances of success compared with traditional approaches by enabling the testing of a very large (unprecedented) number of plasma biomarker candidates. Aim 1. Using SILAC-labeled mice, identify proteins induced in the bone marrow in response to radiation, and subsequently test hundreds of these putative tissue injury markers to identify the subset that are stably elevated in the plasma post-exposure. Aim 2: Characterize candidate marrow injury biomarkers identified in Aim 1 with respect to their: a. stability in plasma over time, dose range, and dose rates b. correlation with clinical endpoints (indicators of hematopoietic syndrome) c. specificity for damage to the hematopoietic system and damage caused by radiation d. use across a heterogeneous population (pediatric, geriatric, gender, genetically susceptible) Aim 3. Determine which of the radiation biomarkers of marrow damage identified in the mouse are elevated in human blood following radiation exposure, and develop a point-of-care assay device that will form the basis of subsequent human clinical validation trials (beyond this proposal).

描述（由申请人提供）：当全身辐射暴露在 3-8 Gy 时，死亡的主要原因是造血综合征。通过对受害者进行细胞因子治疗和积极支持治疗的快速分类，许多死亡是可以预防的。不幸的是，目前识别造血综合征风险患者的方法存在不准确、费用高、分析时间长和诊断延迟的问题。此外，这些方法都不能直接测量对骨髓的辐射损伤，也不能表明残余造血的存在，并且大多数方法不适用于紧急情况下的护理点。因此，在放射事件发生后，迫切需要一种可现场部署的诊断仪来识别将出现潜在致命且通常可治疗的造血综合征的受害者。 我们建议建立一个非常成功的临床诊断范例，即受伤的组织渗漏、脱落和/或分泌蛋白质 进入血浆，它们是指示组织损伤程度的有用生物标志物。先例包括心肌梗塞中的肌钙蛋白、肝损伤中的转氨酶、肌肉损伤中的肌酸激酶以及胰腺过程中的脂肪酶。因此，我们假设在辐射引起的骨髓损伤后，蛋白质从骨髓释放到血流中，在血流中它们提供有用的生物标志物信号，预测造血综合征的发作和严重程度。我们建议使用一种创新方法来识别这些辐射引起的骨髓损伤的血浆生物标志物，该方法结合了我们在生物标志物发现管道中开发和验证的靶向蛋白质组技术，通过能够测试大量（前所未有的）血浆生物标志物候选物，与传统方法相比，这将大大增加我们的成功机会。 目标 1. 使用 SILAC 标记的小鼠，鉴定骨髓中响应辐射而诱导的蛋白质，然后测试数百个这些推定的组织损伤标记物，以确定暴露后血浆中稳定升高的子集。目标 2：描述目标 1 中确定的候选骨髓损伤生物标志物的以下特征：血浆随时间、剂量范围和剂量率的稳定性 b．与临床终点（造血综合征指标）的相关性 c．造血系统损伤和辐射引起的损伤的特异性 d．在异质人群（儿童、老年人、性别、遗传易感人群）中使用 目标 3. 确定在小鼠中发现的哪些骨髓损伤的辐射生物标志物在辐射暴露后在人体血液中升高，并开发一种即时检测装置，该装置将构成后续人体临床验证试验的基础（超出本提案）。