Proteomics of silent cerebral infarction biomarker in Sickle Cell Disease

无症状性脑梗塞(SCI)发生在22%的镰状细胞病(SCO)儿童中，与 认知功能下降，是中风发生的重要危险因素。通过 定义，这些儿童有符合脑梗塞的MRI和病理证据，没有明显的症状。 中风的症状。由于脊髓损伤的病因不明，我们缺乏轻松识别儿童脊髓损伤的手段。 对SCI或有SCI危险的患者进行早期干预或跟踪治疗成功。这个项目的总体目标是 GRANT建议确定SCI儿童SCI的诊断/病因血浆生物标记物。在我们的试点中 对患有或不伴有脊髓损伤的患者的血浆样本进行无偏倚蛋白质组学分析 SIT试验中，我们发现脊髓损伤儿童和有脊髓损伤儿童的血浆蛋白谱有显著差异 鉴定了两种循环中的脑特异性蛋白质，神经细胞黏附分子1和胶质纤维酸性 蛋白质，一种已知的中风生物标志物。在这些研究的基础上，我们建议识别血浆 脊髓损伤的生物标志物，确定循环中的生物标志物蛋白是否可以预测脊髓损伤并深入到 脊髓损伤儿童的血浆蛋白质组以确定脑内有效的诊断生物标志物和 脉管系统。本研究的意义在于鉴定脊髓损伤的循环标志物。 将允许对这些儿童进行早期干预，并为新疗法的设计提供洞察力。这是 尤其重要的是，因为我们目前的重复输血疗法本身就具有很高的发病率。 因此，我们假设来自sco和sci儿童的血浆样本中将含有 脊髓损伤的诊断/病因生物标志物。为了检验这一假设，我们的建议借鉴了独特的 SIT试验生物储备库中大量高表型血浆样本的来源和 约翰霍普金斯大学NHLBI心血管蛋白质组学中心。我们将在三个目标中：第一，使用最先进的 无偏倚蛋白质组学技术(高效液相色谱。ITRAQ和LC/MS/MS)，鉴定血浆诊断生物标志物 第二，结合蛋白质印迹和高通量多重分析来验证我们的生物标记物 针对SIT试验血浆库的抗体阵列，以测试生物标记物在预测 第三，回到大脑，将生物标记蛋白定位到大脑，并测试它们对 SCO与缺血性神经损伤。 这些研究描述了发现和验证阶段的工作，以揭示诊断/病因 儿童脊髓损伤的血浆生物标志物。这些研究也有可能提供新的 对上海合作组织中风和中风总体发展的洞察。

Silent cerebral infarctions (SCI) occur in 22% of children with sickle cell disease (SCO), are associated with decreased cognitive function and are a significant risk factor for the development of stroke. By definition, these children have MRI and pathologic evidence consistent with cerebral infarction, without overt symptoms of stroke. As the etiology of SCI is unknown, we lack the means to easily identify children with SCI or at risk for SCI to guide early intervention or follow the success of therapy. The overall goal of this grant proposal is to identify diagnostic/etiologic plasma biomarkers of SCI in children with SCO. In our pilot studies, using a non-biased proteomic analysis of plasma samples from patients with or without SCI in the SIT Trial, we have found significant differences in the plasma protein profile of children with SCI and have identified two circulating brain specific proteins, neural cell adhesion molecule 1 and glial fibrillary acidic protein, a known biomarker of stroke. Based upon these studies, we propose to identify the plasma biomarkers of SCI, establish whether circulating biomarker proteins can predict SCI and probe deep in the plasma proteome of children with SCI to identify diagnostic biomarkers validated to the brain and vasculature. The significance of the proposed studies is that identification of circulating markers of SCI would allow early intervention in these children and provide insight into the design of new therapies. This is particularly important, as our present therapy of repeat transfusions carries a high morbidity in and of itself. Therefore, we hypothesize that plasma samples from children with SCO and SCI will contain diagnostic/etiologic biomarkers of SCI. To examine this hypothesis, our proposal draws on the unique resources of the large and highly phenotyped plasma samples in the SIT Trial Biologic Respository and the NHLBI Cardiovascular Proteomics Center at Johns Hopkins. In three aims we will: first, using state of the art non-biased proteomic techniques (HPLC. ITRAQ and LC/MS/MS), identify plasma diagnostic biomarkers of SCI; second, validate our biomarkers using a combination of western blotting and high throughput multiplex antibody arrays against the SIT Trial plasma repository to test the efficacy of the biomarkers in predicting SCI; and third, go back to the brain, to localize biomarker proteins to the brain and test their specificity for SCO and ischemic neurologic injury. These studies describe the discovery and validation phases of work to uncover diagnostic/etiologic plasma biomarkers of SCI in children with SCO. It is also possible that these studies may provide new insights into the development of stroke in SCO and stroke in general.