Understanding ischemia in children with tuberculous meningitis (iThemba)

项目总结 背景：每年有一百万儿童患上结核病，其中四分之一死亡。结核性脑膜炎 (TBM)是最严重的结核病，即使诊断和治疗，20%的人死亡，超过50%的人 幸存者留下的是永久性的神经残疾。与TBM相关的大部分发病率和死亡率 是由于梗塞引起的大小血管炎症和血栓形成。尽管如此，我们的 对脑梗塞发病机制的认识还很有限，尤其是在儿童。我们假设 血管内皮细胞和血浆促血栓形成和抗血栓机制的失衡，炎症途径和 血管增殖过程是脑梗塞发生的基础。我们的团队有着广泛的记录 研究团队由临床领域的世界领先者组成 流行病学、蛋白质组学、转录组学、生物信息学、神经科学和放射学。 方法：我们将招募80名30个月以上可能或确诊为TBM的儿童，并获取样本。 血液和脑脊液(CSF)。所有儿童在基线和2周时接受MRI和FDG PET/CT检查 他们将进行重复的核磁共振检查，并收集更多的血液和脑脊液样本。然后在24周时进行核磁共振检查 将在48周时进行神经发育评估。神经成像将量化儿童之间的差异 有无脑梗塞和影像与临床表现和预后的关系。它还将识别半暗带 预示未来脑梗塞发展/演变的区域。我们将用以下样品丰富实验室分析： 在2016至2020年间招募的50名可能患有或确诊为结肠炎的儿童。血液和血液的RNA测序 脑脊液将被用来识别差异表达的基因和确定 有和没有脑梗塞的儿童，以及在基线和2周时采集的样本之间的差异。目标明确 将对血浆和脑脊液进行免疫分析和发现质谱分析，以确定差异 在蛋白质丰度方面，重点放在参与凝血和内皮功能的蛋白质上。最后，我们会 整合转录组学、蛋白质组学和放射组学，以产生对 脑瘫患儿脑梗塞的发病机制。我们的目标是将孩子们分成几个生物学/解剖学的 表型，每一种可能受益于不同的治疗方法。然后我们将探索，使用 计算机模拟，治疗干预对每种表型生物通路的影响。我们 预计这项工作可以为开发可以分层治疗的护理点测试铺平道路 在确诊的时候。 影响：更全面地了解TBM儿童脑梗塞的病理生理学将 允许有针对性的宿主导向疗法，有可能缓解或消除这一后果 毁灭性的状况。

PROJECT SUMMARY Background: One million children develop tuberculosis (TB) each year and a quarter of these die. TB meningitis (TBM) is the most severe form of TB disease and even if diagnosed and treated, 20% die and over 50% of survivors are left with permanent neurological disability. Much of the morbidity and mortality associated with TBM is due to infarction caused by large and small vessel inflammation and thrombosis. Despite this, our understanding of the pathogenesis of infarction in TBM is limited, especially in children. We hypothesize that imbalance of the endothelial and plasma pro- and anti-thrombotic mechanisms, inflammatory pathways and vascular proliferative processes underlies cerebral infarction in TBM. Our group has an extensive track record of clinical research into children with TBM and the research team consists of world leaders in the fields of clinical epidemiology, proteomics, transcriptomics, bioinformatics, neurosciences, and radiology. Methods: We will recruit 80 children with probable or confirmed TBM over 30 months and obtain samples of blood and cerebrospinal fluid (CSF). All children will undergo MRI and FDG PET/CT at baseline and at 2 weeks they will have repeat MRI with further blood and CSF samples collected. MRI will then be carried out at 24 weeks will neurodevelopmental assessment at 48 weeks. The neuroimaging will quantify differences between children with and without infarction and relate imaging to clinical presentation and outcome. It will also identify penumbral regions indicating future infarct development/evolution. We will enrich the laboratory analyses with samples from 50 children with probable or confirmed TBM, recruited between 2016 and 2020. RNA sequencing of blood and CSF will be used to identify differentially expressed genes and identify implicated biological pathways between children with and without infarction and between samples taken at baseline and at 2 weeks. Targeted immunoassays and discovery mass spectrometry will be performed on plasma and CSF to determine differences in protein abundance, with a focus on proteins involved in coagulation and endothelial function. Finally, we will integrate the transcriptomics, proteomics and radiomics to generate a comprehensive understanding of the pathogenesis of infarction in children with TBM. We aim to group children into several biological/anatomical phenotypes, each of which may benefit from a different therapeutic approach. We will then explore, using computer simulation, the impact of therapeutic interventions on biological pathways in each phenotype. We anticipate that this work could pave the way for the development of point-of-care tests that could stratify therapy at the time of diagnosis. Impact: A more comprehensive understanding of the pathophysiology of infarction in children with TBM would permit targeted host-directed therapies, with the potential to moderate or eliminate the consequences of this devastating condition.