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: 8370399
• 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/8370399
• 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; Liver; 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; 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). PUBLIC HEALTH RELEVANCE: History tells us that amidst the immediate panic of a nuclear attack/accident in an urban area, tens-to-hundreds of thousands of panicked people demanding to be evaluated for exposure will likely overwhelm our emergency care system and consequently jeopardize effective triage and treatment of those whose lives are imminently in danger. Additionally, our current tests for determining a specific victim's level of exposure are inadequate to ensure that every victim gets rapid and ideal medical treatment for radiation sickness, which is often survivable if treated rapidly and aggressively. In this proposal, we will develop a field-deployable blood test that will allow us to rapidly determine whether an individual has or has not been exposed to radiation, and if so, what is the best course of treatment to ensure the highest chance of survival.

描述（由申请人提供）：在3-8 Gy的总体辐射暴露时，死亡的主要原因是造血综合征。这些死亡中的许多人可以通过快速的受害者进行细胞因子疗法和积极的支持护理来预防。不幸的是，目前用于识别有造血综合征风险的患者的方式遭受了不准确，高费用，较长的分析时间和延迟诊断的患者。此外，这些方法均未直接测量对骨髓的辐射损害，也不表明存在残留的造血症，并且大多数人不适合在紧急情况下进行护理。因此，在放射学事件后，识别出可能会发展可能致命的造血综合征的受害者，对使用野外诊断的诊断非常不满意。 我们建议建立在临床诊断中非常成功的范式上，即受伤的组织泄漏，脱落和/或分泌蛋白质 进入血浆，它们是有用的生物标志物，表明组织损伤的程度。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液中，在那里它们提供了有用的生物标志物信号，可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的方法来识别辐射诱导的骨髓损伤的这些等离子体生物标志物，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有针对性蛋白质组学技术，这些技术将大大增加与传统方法相比的成功机会，从而实现我们的成功机会。非常大的（前所未有的）等离子生物标志物候选者。 AIM 1。使用SILAC标记的小鼠，鉴定响应辐射中在骨髓中诱导的蛋白质，然后测试数百种推定的组织损伤标记，以鉴定暴露后血浆中稳定升高的子集。 AIM 2：特征候选骨髓损伤生物标志物在AIM 1中鉴定出相对于其：a。随着时间的推移，剂量范围和剂量率b。等离子体的稳定性b。与临床终点（造血综合征指标）c的相关性。对造血系统损害的特异性和辐射d造成的损害。在异质种群中使用（小儿，老年，性别，遗传易感性）AIM 3。确定在辐射后，在小鼠中鉴定出的小鼠骨髓损伤的辐射生物标志物在辐射后升高，并开发出疗程的分析设备这将构成随后的人类临床验证试验的基础（超出此提案）。 公共卫生相关性：历史告诉我们，在城市地区的核攻击/事故的立即恐慌中，数以万计的恐慌的人要求进行暴露，可能会使我们的紧急护理系统不堪重负，因此危害有效的有效性分类和待遇的人天生有危险。此外，我们目前确定特定受害者暴露水平的测试不足以确保每个受害者对放射病的迅速和理想的医疗治疗，如果经过迅速而积极的治疗，通常可以生存。在此提案中，我们将开发一项可采用的血液测试，这将使我们能够迅速确定一个人是否是否 已经或尚未暴露于辐射，如果是的话，确保生存机会最高的最佳治疗方法是什么。