CEREBRAL MALARIA-INDUCED APOPTOSIS AND BRAIN PATHOLOGY

脑疟疾引起的细胞凋亡和脑病理学

• 批准号: 7561418
• 负责人: Jonathan K. Stiles
• 金额: $ 14.65万
• 依托单位: MOREHOUSE SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-06-01 至 2008-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7561418
• 关键词: Antibodies; Apoptosis; Apoptotic; Biological Assay; Blocking Antibodies; Brain; Brain Injuries; Brain Pathology; Cell Line; Cerebral Malaria; Child; Cognition; Computer Retrieval of Information on Scientific Projects Database; DNA; Data; Endothelial Cells; Funding; Gene Proteins; Gene Silencing; Genes; Genomics; Goals; Grant; Human; Imaging Techniques; In Situ Nick-End Labeling; In Vitro; Institution; Malaria; Mass Spectrum Analysis; Mediating; Methods; Modeling; Mus; Neuroglia; Neurologic; Neurons; Numbers; Parasites; Pathology; Patients; Plasmodium; Platelet Activating Factor; Procedures; Proteins; Proteomics; RNA Interference; Research; Research Personnel; Resources; Rodent; Role; Source; Survivors; Therapeutic; Two-Dimensional Gel Electrophoresis; United States National Institutes of Health; brain tissue; doxorubicin/fluorouracil/melphalan protocol; killings; knock-down; outcome forecast; platelet activating factor receptor; prevent; protein expression

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Specific Aims Malaria (P. falciparum) infects 200 to 300 million people globally and kills 2 million (mostly children) every year. 15% of fatal cases are due to cerebral malaria (CM) and other severe forms of malaria. A significant segment of CM patients die regardless of recommended treatment. A significant number of survivors develop neurological complications and cognition problems. The precise mechanisms responsible for CM induced brain damage and poor prognosis is unclear. The main hypothesis of this project is that Plasmodium apoptotic factor(s) (PAF) induce neuronal and microvascular endothelial cell apoptosis and that selectively blocking PAFmediated apoptosis will negate or significantly reduce apoptosis and CM-induced pathology. Our objective is to identify and characterize the role of PAF in CM-induced brain pathology using human brain endothelial (HBVEC), glial, and neuronal cell lines, as well as our established rodent CM model respectively. We plan to utilize genomics, proteomics, immunological methods, imaging techniques, ultrastructural analysis, and targeted gene inactivation (RNA interference, RNAi) to pursue these goals. Our long-term goal is to target PAF as a therapeutic strategy for preventing or treating CM-induced brain pathology. Three Specific Aims have been proposed to study the role of apoptosis in CM-induced brain pathology. Specific aim 1 will compare and contrast CM induced apoptosis in murine CM models with that in P. falciparum-induced human CM using post mortem brain tissues. CM-induced expression changes in apoptotic factors in murine and human CM will be compared by gene and protein expression analysis, ultrastructure, immunohistology, and DNA cleavage (TUNEL) analysis. Specific aim 2 will identify and characterize the key apoptotic factor(s) (PAF) mediating CM-induced apoptosis. Gene and protein expression data (gene microarray chips, 1D & 2D gel electrophoresis) and protein Mass spectroscopy (MALDI-TOF) will be utilized to identify and characterize the potential role of PAF in CM-induced apoptosis in HBVEC, glia, and neuronal cell lines in vitro. Specific aim 3 will functionally characterize CM-mediated apoptosis in CM. Antibody blocking procedures and, if necessary, RNA interference (RNAi) strategies will be used to knock down expression of host PAF receptor(s) in HBVEC, glial, and neuronal cell lines induced by parasite infected RBC (IRBC) and uninfected controls and assayed for apoptosis. BBB integrity will be evaluated in an in vitro BBB model in the presence or absence of IRBC. We will also exogenously block PAF with anti-PAF antibody in murine CM model and examine effects on BBB and CM pathology.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 具体目标 疟疾（恶性疟原虫）每年感染全球2亿至3亿人，并导致200万人死亡（其中大部分是儿童）。15%的致命病例是由于脑型疟疾（CM）和其他严重形式的疟疾。 无论推荐的治疗如何，CM患者的显著部分死亡。相当多的幸存者出现神经系统并发症和认知问题。 CM引起脑损伤和预后不良的确切机制尚不清楚。 该项目的主要假设是疟原虫凋亡因子（PAF）诱导神经元和微血管内皮细胞凋亡，并且选择性地阻断PAF介导的凋亡将否定或显著减少凋亡和CM诱导的病理。 我们的目的是确定和表征PAF在CM诱导的脑病理学中的作用，分别使用人脑内皮细胞（HBVEC），神经胶质细胞和神经元细胞系，以及我们建立的啮齿动物CM模型。我们计划利用基因组学、蛋白质组学、免疫学方法、成像技术、超微结构分析和靶向基因失活（RNA干扰，RNAi）来实现这些目标。 我们的长期目标是靶向PAF作为预防或治疗CM诱导的脑病理学的治疗策略。已经提出了三个特定的目的来研究细胞凋亡在CM诱导的脑病理中的作用。 具体目标1将使用死后脑组织比较和对比鼠CM模型中CM诱导的细胞凋亡与恶性疟原虫诱导的人CM中的细胞凋亡。将通过基因和蛋白质表达分析、超微结构、免疫组织学和DNA裂解（TUNEL）分析比较CM诱导的鼠和人CM中凋亡因子表达变化。 具体目标2将鉴定和表征介导CM诱导的细胞凋亡的关键凋亡因子（PAF）。基因和蛋白质表达数据（基因微阵列芯片，1D和2D凝胶电泳）和蛋白质质谱（MALDI-TOF）将用于识别和表征PAF在体外HBVEC，神经胶质细胞和神经元细胞系中CM诱导的凋亡中的潜在作用。 具体目标3将在功能上表征CM中CM介导的细胞凋亡。将使用抗体阻断程序和（如有必要）RNA干扰（RNAi）策略来敲低由寄生虫感染的RBC（IRBC）和未感染对照诱导的HBVEC、神经胶质细胞和神经元细胞系中宿主PAF受体的表达，并测定细胞凋亡。 将在存在或不存在IRBC的情况下在体外BBB模型中评价BBB完整性。我们还将在小鼠CM模型中用抗PAF抗体外源性阻断PAF，并检查对BBB和CM病理学的影响。