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）： 当前对HIV感染患者进行抗逆转录病毒治疗（CART）的治疗可有效抑制病毒复制，预防免疫抑制和寿命增加。尽管有这些进展，即使在卡车存在下，也无法完全预防慢性神经炎症，从而导致与HIV相关的神经认知障碍（Hand）的发展。上述临床观察结果促使定量生物标志物有必要帮助检测和监测HIV感染引起的中枢神经系统（CNS）并发症。血脑屏障（BBB）的功能障碍是HIV-1神经病发生的众所周知现象，可能导致神经变性。我们提出了反映BBB损伤的新型生物标志物，即对含有从脑内皮释放的紧密连接蛋白（TJP）的外卢尔小泡（EMV）检测。我们发现，由于炎症和血管重塑而发生EMV从脑内皮中释放。重要的是，对脑内皮细胞（EC）的同样炎症性侮辱会诱导屏障的渗透性和生化变化对紧密连接络合物的变化。该项目在生物标志物发现的创新工具的开发和鉴定在BBB处发生的新型病理生理现象的创新工具都是独一无二的。在第一个目标中，我们将在时间上介绍并按时间学定义EMV中TJP的存在，这是由于相关炎症刺激和HIV病毒毒素所致。这些确定将与测量屏障完整性并联做出，从而允许EMV脱落与BBB功能障碍之间的相关性。由于结构蛋白鉴定了EMV的细胞来源，因此第二个目标探讨了携带TJP作为BBB功能障碍血清学生物标志物的基础的EMV概念。使用最敏感的ELISA可用技术，我们构建了原型ELISA，以测量含有TJP的血液循环EMV的水平。使用来自HIV-1感染个体的患者血清，我们提供了概念证明的结果，这种新型生物标志物与手的诊断相关。在第三个目标中概述的实验剖析了与脑EC微囊泡形成和TJP包装的生物发生有关的合理分子机制。它将不仅能够理解如何触发EMV形成，而且还将揭示细胞内的靶标，以促进BBB保护的药理干预措施。