Acute brain injury and blood-brain barrier dysfunction in cerebral malaria (ABC)

脑型疟疾（ABC）中的急性脑损伤和血脑屏障功能障碍

基本信息

批准号：
10433313
负责人：
Scott G Canfield
金额：
$ 26.64万
依托单位：
INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-02-15 至 2024-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10433313
关键词：
Acidosis Acute Acute Brain Injuries Admission activity Alzheimer&apos s Disease Animal Model Animals Astrocytes Axon Binding Biological Markers Blood Blood - brain barrier anatomy Blood Circulation Blood Vessels Brain Brain Injuries Cell Adhesion Cell Adhesion Molecules Cells Cerebral Malaria Cerebrospinal Fluid Characteristics Child Childhood Clinical Coma Complication Data Endothelium Environment Erythrocytes Evaluation Event Falciparum Malaria Functional disorder Future Glial Fibrillary Acidic Protein Glucose Goals Hemolysis Human Impact evaluation Impaired cognition In Vitro Infection Inflammation Injury Knowledge Lactate Dehydrogenase Mediating Metabolic Modeling Nervous System Trauma Neurocognitive Neuroglia Neurologic Neuronal Injury Neurons Parasites Pathogenesis Pathogenicity Pathway interactions Pediatric cohort Physiological Plasma Plasmodium falciparum Prognostic Marker Property Proteins Research Sampling Survivors Testing Tight Junctions Virulence Factors Work blood-brain barrier crossing blood-brain barrier disruption brain cell brain endothelial cell brain parenchyma cell injury cerebral microvasculature cognitive testing cohort deprivation disability endothelial dysfunction follow-up hypoxic ischemic injury in vitro Model induced pluripotent stem cell insight monolayer mortality nervous system disorder neuronal cell body neuropathology new therapeutic target novel marker prevent stem cell model tau Proteins tau-1

项目摘要

ABSTRACT Cerebral malaria (CM) is an acute neurologic complication of infection with Plasmodium falciparum malaria that presents clinically as an unarousable coma. In CM, several pathogenic mechanisms interact to cause the characteristic neurocognitive sequelae observed in a quarter of surviving children. Infected erythrocytes sequester in the brain microvasculature leading to widespread inflammation, endothelial activation, hypoxic/ischemic injury, glucose abnormalities, hemolysis and cellular injury, resulting blood-brain barrier (BBB) dysfunction, which may interact to cause brain injury. We have found that elevated tau and other markers of injury to neurons are elevated in blood circulation and associated persistent neurodisability in survivors of CM. Further, these brain injury markers are associated with factors involved in the pathogenesis of CM, including glucose abnormalities, cellular injury, and endothelial/BBB dysfunction. What remains unclear is how intraerythrocytic parasites confined within the vascular space, cause injury to neurons without crossing the BBB. Thus, there is a critical need for mechanistic studies to define interactions between factors involved in the pathogenesis of CM leading to BBB dysfunction and neuronal injury. Animal and in-vitro BBB models have been key in advancing our knowledge of CM pathogenesis but limitations of existing models include: 1) interspecies variability of non-falciparum animal models, 2) use of endothelial monolayers that lack key components of the brain parenchyma, 3) reliance on immortalized or primary brain microvascular endothelial cell (BMECs) that lack physiologically relevant barrier properties and/or suffer from batch-to-batch variability. To overcome these limitations, we have developed a multicellular BBB model comprised of human-derived BMECs, neurons, and astrocytes. Using our multicellular BBB model, we propose to investigate neuronal injury and examine interactions between factors involved in CM pathogenesis and biomarkers of brain injury. And we will validate our brain injury biomarker data in a follow-up cohort of children with CM tested for cognitive impairments at 12- months. We hypothesize that sequestration of IEs to BMECs results in BBB dysfunction and the subsequent decrease of glucose availability and release of cellular injury marker lactate dehydrogenase (LDH) causing neuronal injury, which is a predictor of persistent neurodisability in clinical CM. Our hypothesis will be testing by the following specific aims: 1) evaluation of the impact of glucose deprivation, cellular injury, and endothelial dysfunction on neuronal injury in an in vitro human-derived multicellular BBB model, and 2) evaluation of a panel of brain injury biomarkers as predictors of neurodisability in clinical CM. Upon completion, this work will establish our human-derived multicellular BBB model as the standard for investigating mechanisms underlying neuronal injury, which we have shown to be a predictor of persistent neurodisability in clinical CM. Our model has the potential to transform the field of in-vitro CM neuropathology by facilitating research into new therapeutic targets to prevent or reduce future neurodisability after pediatric CM.

抽象的脑型疟疾（CM）是恶性疟原虫疟疾感染的一种急性神经系统并发症，临床上表现为无法唤醒的昏迷。在 CM 中，多种致病机制相互作用，导致在四分之一的幸存儿童中观察到特征性神经认知后遗症。受感染的红细胞隔离在大脑微血管系统中，导致广泛的炎症、内皮激活、缺氧/缺血性损伤、血糖异常、溶血和细胞损伤，导致血脑屏障 (BBB) 功能障碍，可能相互作用导致脑损伤。我们发现 tau 蛋白和其他标记物升高 CM 幸存者的血液循环中神经元损伤加剧，并伴有持续性神经功能障碍。此外，这些脑损伤标志物与 CM 发病机制相关的因素有关，包括葡萄糖异常、细胞损伤和内皮/血脑屏障功能障碍。目前尚不清楚的是如何红细胞内寄生虫局限于血管间隙内，在不穿过血脑屏障的情况下对神经元造成损伤。因此，迫切需要进行机制研究来定义参与因素之间的相互作用。 CM 的发病机制导致 BBB 功能障碍和神经元损伤。动物和体外 BBB 模型提高我们对 CM 发病机制认识的关键，但现有模型的局限性包括：1）种间非恶性疟原虫动物模型的变异性，2) 使用缺乏恶性疟原虫关键成分的单层内皮细胞脑实质，3）依赖于缺乏的永生化或原代脑微血管内皮细胞（BMEC）生理相关的阻隔性能和/或存在批次间差异。为了克服这些由于局限性，我们开发了一种多细胞 BBB 模型，由人源 BMEC、神经元和星形胶质细胞。使用我们的多细胞 BBB 模型，我们建议研究神经元损伤并检查 CM 发病机制相关因素与脑损伤生物标志物之间的相互作用。我们将验证我们在 CM 儿童随访队列中的脑损伤生物标志物数据在 12 岁时进行了认知障碍测试几个月。我们假设 IE 与 BMEC 的隔离导致 BBB 功能障碍以及随后的葡萄糖利用率的降低和细胞损伤标志物乳酸脱氢酶（LDH）的释放导致神经元损伤，这是临床 CM 中持续性神经功能障碍的预测因子。我们的假设将通过以下方式进行检验以下具体目标：1）评估葡萄糖剥夺、细胞损伤和内皮细胞的影响体外人源多细胞 BBB 模型中神经元损伤的功能障碍，以及 2) 小组评估脑损伤生物标志物作为临床 CM 神经功能障碍的预测因子。这项工作完成后，将建立我们的人源多细胞 BBB 模型作为研究神经元潜在机制的标准损伤，我们已证明它是临床 CM 中持续性神经功能障碍的预测因子。我们的模型有通过促进新治疗靶点的研究，有可能改变体外 CM 神经病理学领域预防或减少儿科 CM 后未来的神经功能障碍。