Proteomics of silent cerebral infarction biomarker in Sickle Cell Disease

无症状性脑梗死（SCI）发生在22%的镰状细胞病（SCO）儿童中， 认知功能下降，是发生中风的重要危险因素。通过 定义，这些儿童有MRI和病理证据符合脑梗死，没有明显的 中风的症状由于SCI的病因不明，我们缺乏容易识别儿童的方法。 SCI或处于SCI风险中的患者，以指导早期干预或跟踪治疗的成功。这个项目的总体目标是 基金提案是鉴定SCO儿童SCI的诊断/病因学血浆生物标志物。在我们的试播集里 研究，使用无偏倚的蛋白质组学分析血浆样本，从患者或没有SCI，在 SIT试验中，我们发现SCI儿童的血浆蛋白质谱存在显著差异， 鉴定了两种循环脑特异性蛋白，神经细胞粘附分子1和胶质细胞酸性蛋白， 蛋白质，一种已知的中风生物标志物。基于这些研究，我们建议鉴定血浆 SCI的生物标志物，确定循环生物标志物蛋白是否可以预测SCI并深入探讨 脊髓损伤儿童血浆蛋白质组，以确定诊断生物标志物验证大脑， 脉管系统这些研究的意义在于确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见解。这是 特别重要的是，我们目前的重复输血疗法本身具有很高的发病率。 因此，我们假设SCO和SCI儿童的血浆样本中含有 SCI的诊断/病因学生物标志物。为了检验这一假设，我们的建议借鉴了独特的 在SIT试验生物储存库和 约翰霍普金斯NHLBI心血管蛋白质组学中心。我们将有三个目标：第一，利用最先进的技术 非偏倚蛋白质组学技术（HPLC. ITRAQ和LC/MS/MS），鉴定血浆诊断生物标志物， SCI;第二，使用蛋白质印迹和高通量多重检测的组合来验证我们的生物标志物 针对SIT试验血浆储存库的抗体阵列，以测试生物标志物在预测 第三，回到大脑，将生物标志物蛋白定位到大脑，并测试它们对脊髓损伤的特异性。 SCO与缺血性神经损伤 这些研究描述了工作的发现和验证阶段，以揭示诊断/病因 SCO儿童SCI的血浆生物标志物。这些研究也可能提供新的 对SCO中卒中和一般卒中发展的见解。

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.