"PATHOGENIC MECHANISMS AND THERAPUETIC STRATEGIES IN MELAS"

“MELAS 的致病机制和治疗策略”

• 批准号: 7893436
• 负责人: MERCY MASCREEN DAVIDSON
• 金额: $ 32.8万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2015-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7893436
• 关键词: Address; Adherens Junction; Animal Model; Astrocytes; Autopsy; Biochemical; Biological Assay; Blood - brain barrier anatomy; Blood Vessels; Brain; Capillary Endothelial Cell; Cell Culture Techniques; Cells; Cerebral Edema; Cerebral Infarction; Chemicals; Coculture Techniques; Collaborations; Collagen; Data; Defect; Diffusion weighted imaging; Disease; Edema; Electrical Resistance; Endothelial Cells; Etiology; Event; Functional disorder; Funding; Genes; Goals; Human; Hydrocortisone; Immunoblotting; In Vitro; Inflammatory; Intercellular Junctions; Interferons; Inulin; Ions; Laboratories; Lactate Transporter; Lactic Acidosis; Lead; Leucine-Specific tRNA; Measurement; Measures; Mediating; Membrane; Methods; Micropore Filters; Mitochondria; Mitochondrial DNA; Mitochondrial Diseases; Mitochondrial Encephalomyopathies; Modeling; Molecular; Mutation; Pathogenesis; Patients; Permeability; Pharmaceutical Preparations; Physiology; Play; Point Mutation; Process; Protective Agents; Proteins; Regulation; Relative (related person); Reporting; Research Personnel; Respiratory Chain; Role; Side; Steroids; Stroke; Sucrose; System; Testing; Therapeutic; Therapeutic Agents; Therapeutic Intervention; Tight Junctions; Tissues; Translating; Vascular Permeabilities; Water; Work; base; bench to bedside; brain tissue; cadherin 5; fetal; fighting; flexibility; follow-up; glycerylphosphoinositol; immunocytochemistry; in vitro Model; in vivo; insight; mind control; mitochondrial DNA mutation; mitochondrial dysfunction; occludin; programs; solute; therapeutic target; time interval; tissue culture; water channel

MELAS, a common mitochondrial disease associated with a m!3243A>G mutation in the tRNALeu(UUR) gene of the mitochondrial DNA, is characterized by strokes, lactic acidosis, cerebral infarction and edema. The pathogenesis of strokes and edema in MELAS is not understood. Without a proper understanding of the pathophysiology, it has not been possible to devise rational therapies for this devastating disease. Our hypothesis is that mitochondrial dysfunction in the cortical blood vessels causes a breakdown of the blood-brain barrier (BBB) leading to strokes and associated edema. We have successfully constructed a working in vitro model of a normal and MELAS BBB. We propose to study the function of the BBB in MELAS in this model. Specifically, we will analyze tight junction proteins that regulate BBB permeability, water channels, and lactate transporters and correlate our results with respiratory chain function. These in vitro studies at the cellular level will be extended to MELAS brain sections to compare and confirm our findings at the tissue level. Therapeutic management of MELAS is anecdotal and generally ineffective. Our in vitro model is a dynamic system that can be manipulated to test various BBB protective drugs, which can restore barrier function and thus can be useful therapeutic agents for strokes, lactic acidosis, and associated edema. We will use this model to test the effect of steroids in regulating water channels and of glycerphosphoinositol and interferon-b-1a in modulating the inflammatory process after strokes. First, these studies will provide insight into the physiology of the BBB. Secondly, they will reveal changes in vascular permeability in MELAS due to mitochondrial dysfunction and energy shortage. Third, the culture model will allow us to evaluate the sequence of events leading to the increase in permeability. Finally, the in vitro model provides a flexible system to test the therapeutic value of drugs to be used this devastating illness.

MELAS 是一种常见的线粒体疾病，与线粒体 DNA 的 tRNALeu(UUR) 基因 m!3243A>G 突变相关，其特征是中风、乳酸性酸中毒、脑梗塞和水肿。 MELAS 中风和水肿的发病机制尚不清楚。如果没有对病理生理学的正确理解，就不可能为这种毁灭性的疾病设计出合理的疗法。我们的假设是，皮质血管中的线粒体功能障碍会导致血脑屏障（BBB）崩溃，从而导致中风和相关水肿。我们已经成功构建了正常和 MELAS BBB 的体外工作模型。我们建议在此模型中研究 MELAS 中 BBB 的功能。具体来说，我们将分析调节血脑屏障通透性、水通道和乳酸转运蛋白的紧密连接蛋白，并将我们的结果与呼吸链功能相关联。这些细胞水平的体外研究将扩展到 MELAS 脑切片，以比较和证实我们在组织水平的发现。 MELAS 的治疗管理是轶事且通常无效。我们的体外模型是一个动态系统，可用于测试各种 BBB 保护药物，这些药物可以恢复屏障功能，因此可以成为中风、乳酸性酸中毒和相关水肿的有用治疗剂。我们将使用该模型来测试类固醇在调节水通道中的作用以及甘油磷酸肌醇和干扰素-b-1a 在调节中风后炎症过程中的作用。 首先，这些研究将深入了解血脑屏障的生理学。其次，他们将揭示由于线粒体功能障碍和能量短缺而导致的 MELAS 血管通透性的变化。第三，培养模型将使我们能够评估导致渗透性增加的事件顺序。最后，体外模型提供了一个灵活的系统来测试用于这种毁灭性疾病的药物的治疗价值。