Functional Genomics of the Blood-Brain Barrier

血脑屏障的功能基因组学

• 批准号: 6798689
• 负责人: ERIC V SHUSTA
• 金额: $ 14.34万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-01 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6798689
• 关键词: AIDS dementia complex; astrocytes; bioengineering /biomedical engineering; biotechnology; blood brain barrier; brain cell; cell cell interaction; ethanol; functional /structural genomics; gene expression; human immunodeficiency virus; intermolecular interaction; laboratory rat; macrophage; microarray technology; mixed tissue /cell culture; model; model design /development; neurons; nitric oxide synthase; phenotype; physical chemical interaction; subtraction hybridization; technology /technique development; tissue /cell culture; vascular endothelium

DESCRIPTION (provided by applicant): HIV encephalitis and AIDS dementia complex are neurological disorders that continue to afflict AIDS patients even in the presence of highly active retroviral therapies. Research has suggested that ethanol may exacerbate these symptoms and may do so by modulating the susceptibility of the blood-brain barrier (BBB) to infiltration by HIV infected monocytes or free HIV virus. In order to study these complex interactions, an in vitro model that accurately represents the in vivo BBB is required. When brain microvascular endothelial cell are cultured in vitro, however, they undergo a great deal of de-differentiation and lose many of the specialized biochemical and morphological features observed in vivo. The overall goal of the proposed research is to gain a sophisticated genome-wide understanding of the determinants of the in vivo BBB phenotype that are lost in vitro and progress towards truly representative in vitro BBB models by recreating the in vivo environment. Functional genomics in the form of suppression subtractive hybridization will be used to elucidate the differential gene expression profiles between intact brain microvessels (BBB) and primary cultures of brain microvessel endothelial cells. This will allow the identification of functional clusters of known and novel genes that elicit the unique in vivo BBB phenotype. Armed with this global functional profile, a gene microarray will be created and used as a diagnostic for the quantitative assessment of an in vitro model's ability to reestablish in vivo conditions. Perivascular brain cells can restore some BBB properties when co-cultured with brain microvascular endothelial cells in vitro. The contributions of astrocytes, neurons, and pericytes on the in vitro phenotype will be assessed using the diagnostic gene microarray. Molecular pathways and physiological characteristics that are reestablished by co-culture will be identified and provide targets for rational tuning of in vitro models. Finally, the effects of ethanol on the functional features of the in vitro model will be investigated as a preliminary validation of in vitro models for the study of the complicated interactions between HIV, ethanol, and the BBB.

描述（由申请人提供）： HIV脑炎和艾滋病痴呆综合症是神经系统疾病，即使在存在高效逆转录病毒疗法的情况下，它们仍继续困扰着艾滋病患者。研究表明，乙醇可能会加剧这些症状，并且可能是通过调节血脑屏障 (BBB) 对 HIV 感染的单核细胞或游离 HIV 病毒渗透的敏感性来实现的。为了研究这些复杂的相互作用，需要一个准确代表体内 BBB 的体外模型。然而，当脑微血管内皮细胞在体外培养时，它们会经历大量的去分化，并失去许多在体内观察到的特殊生化和形态特征。拟议研究的总体目标是对体外丢失的体内 BBB 表型决定因素获得复杂的全基因组理解，并通过重建体内环境向真正具有代表性的体外 BBB 模型迈进。 抑制消减杂交形式的功能基因组学将用于阐明完整脑微血管（BBB）和脑微血管内皮细胞原代培养物之间的差异基因表达谱。这将允许识别已知和新基因的功能簇，从而引发独特的体内 BBB 表型。有了这种全局功能图谱，将创建基因微阵列并将其用作体外模型重建体内条件能力的定量评估的诊断方法。血管周围脑细胞与脑微血管内皮细胞在体外共培养时可以恢复部分血脑屏障特性。将使用诊断基因微阵列评估星形胶质细胞、神经元和周细胞对体外表型的贡献。通过共培养重建的分子途径和生理特征将被确定，并为体外模型的合理调整提供目标。最后，将研究乙醇对体外模型功能特征的影响，作为体外模型的初步验证，用于研究 HIV、乙醇和 BBB 之间的复杂相互作用。