Novel serological biomarkers for BBB dysfunction during HIV-1 infection

HIV-1 感染期间 BBB 功能障碍的新型血清学生物标志物

• 批准号: 8659861
• 负责人: Servio Heybert Ramirez
• 金额: $ 38.79万
• 依托单位: TEMPLE UNIV OF THE COMMONWEALTH
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-30 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8659861
• 关键词: AIDS neuropathy; Abbreviations; Biochemical; Biogenesis; Biological Assay; Biological Markers; Blood; Blood - brain barrier anatomy; Brain; Brain imaging; Cell Culture Techniques; Cells; Chronic; Clinical; Complex; Conditioned Culture Media; Coupled; Data; Detection; Development; Diagnosis; Diagnostic; Disease; Electrical Resistance; Endothelial Cells; Endothelium; Enzyme-Linked Immunosorbent Assay; Extracellular Protein; Functional disorder; Guanosine Triphosphate Phosphohydrolases; HIV; HIV Infections; HIV diagnosis; HIV-1; Health; Homeostasis; Immunosuppression; In Vitro; Individual; Infection; Inflammation; Inflammation Mediators; Inflammatory; Injury; Intervention; Laboratories; Longevity; Measurement; Measures; Metric; Molecular; Monitor; NOD/SCID mouse; Nerve Degeneration; Neuraxis; Neurocognitive; Neuroglia; Neurons; Neuropathogenesis; Pathway interactions; Patients; Permeability; Phosphorylation; Process; Proteins; Proteomics; Publishing; Reporting; Risk; Role; Sensitivity and Specificity; Serology; Serum; Sorting - Cell Movement; Source; Specific qualifier value; Stimulus; Structural Protein; System; Technology; Testing; Tight Junctions; Time; Vascular Permeabilities; Vascular remodeling; Virus; Virus Replication; antiretroviral therapy; base; cytokine; design; extracellular; in vivo; innovation; metabolomics; neuroinflammation; novel; prevent; protein complex; protein expression; prototype; public health relevance; research study; tool; virotoxins

DESCRIPTION (provided by applicant): Current treatment of HIV-infected patients with combined antiretroviral therapy (cART) has provided effective suppression of virus replication, prevented immunosuppression and increased lifespan. Despite these advances, even in the presence of cART, chronic neuroinflammation cannot be fully prevented, leading to development of HIV-associated neurocognitive disorders (HAND). The above clinical observation has prompted the necessity for quantitative biomarkers to help in detection and monitoring of central nervous system (CNS) complications resulting from HIV infection. Dysfunction of the blood-brain barrier (BBB) is a well-known phenomenon in HIV-1 neuropathogenesis potentially leading to neurodegeneration. We propose novel biomarkers reflecting BBB injury, namely detection of extracelluar microvesicles (eMVs) containing tight junction proteins (TJP) released from brain endothelium. We discovered that eMV release from the brain endothelium occurs as a result of inflammation and vascular remodeling. Importantly, the same pro- inflammatory insult on brain endothelial cells (EC) induces barrier permeability and biochemical changes to tight junction complexes. This project is unique in both the development of innovative tools for biomarker discovery and identification of a novel pathophysiologic phenomenon occurring at the BBB as consequence of chronic HIV-associated neuroinflammation. In the first aim, we will profile and chronologically define the presence of TJPs in eMV secreted as a consequence of relevant inflammatory stimuli and HIV virotoxins. These determinations will be made in parallel with measurement of barrier integrity, thus allowing for correlations between eMV shedding and BBB dysfunction. Because structural proteins identify the cellular origin of the eMV, the second aim explores the notion of eMV carrying TJP as the basis for a serological biomarker for BBB dysfunction. Using the most sensitive ELISA-based available technology, we have built prototype ELISAs to measure the level of blood circulating eMVs containing TJP. Using patient serum from HIV-1 infected individuals, we provide proof-of concept results that this novel biomarker correlates with diagnosis of HAND. Experiments outlined in the third aim dissect plausible molecular mechanisms involved in the biogenesis of brain EC microvesicle formation and packaging of TJP. It will offer, not only an understanding of how eMV formation is triggered, but also will reveal targets within the cell for pharmacological intervention that promote BBB protection.

描述（由申请人提供）： 目前对艾滋病毒感染者的联合抗逆转录病毒疗法（cART）的治疗可以有效抑制病毒复制，防止免疫抑制并延长寿命。尽管取得了这些进展，但即使存在 cART，也无法完全预防慢性神经炎症，从而导致 HIV 相关神经认知障碍 (HAND) 的发展。上述临床观察提示需要定量生物标志物来帮助检测和监测 HIV 感染引起的中枢神经系统 (CNS) 并发症。血脑屏障 (BBB) 功能障碍是 HIV-1 神经发病机制中的一个众所周知的现象，可能导致神经变性。我们提出了反映血脑屏障损伤的新型生物标志物，即检测含有脑内皮释放的紧密连接蛋白（TJP）的细胞外微泡（eMV）。我们发现大脑内皮释放 eMV 是炎症和血管重塑的结果。重要的是，对脑内皮细胞（EC）的相同促炎性损伤会诱导屏障通透性和紧密连接复合物的生化变化。该项目在开发用于发现生物标志物的创新工具和识别由于慢性 HIV 相关神经炎症而在 BBB 中发生的新型病理生理现象方面都是独一无二的。在第一个目标中，我们将按时间顺序描述和定义由于相关炎症刺激和 HIV 病毒毒素而分泌的 eMV 中 TJP 的存在。这些确定将与屏障完整性的测量同时进行，从而考虑 eMV 脱落和 BBB 功能障碍之间的相关性。由于结构蛋白可识别 eMV 的细胞起源，因此第二个目标是探索携带 TJP 的 eMV 的概念，作为 BBB 功能障碍的血清学生物标志物的基础。使用最灵敏的基于 ELISA 的现有技术，我们构建了 ELISA 原型来测量血液循环中含有 TJP 的 eMV 水平。使用来自 HIV-1 感染者的患者血清，我们提供了概念验证结果，证明这种新型生物标志物与 HAND 的诊断相关。第三个目标中概述的实验剖析了参与脑 EC 微泡形成和 TJP 包装的生物发生的可能分子机制。它不仅将提供对如何触发 eMV 形成的理解，还将揭示细胞内促进 BBB 保护的药物干预目标。