Identification of Biomarkers for BBB Injury-Induced by Drugs and HIV-1 Proteins

药物和 HIV-1 蛋白引起的 BBB 损伤生物标志物的鉴定

• 批准号: 8466253
• 负责人: SHENG-HE HUANG
• 金额: $ 8.1万
• 依托单位: CHILDREN'S HOSPITAL OF LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-02-01 至 2015-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8466253
• 关键词: AIDS neuropathy; Acquired Immunodeficiency Syndrome; Albumins; Alcohols; Area; Biological Markers; Blood; Blood - brain barrier anatomy; Bone Marrow; Brain; Cardiovascular Diseases; Cells; Central Nervous System Diseases; Clinical; Communicable Diseases; Death Rate; Detection; Development; Diagnosis; Disease; Drug abuse; Drug usage; Drug user; Endothelial Cells; Environmental Tobacco Smoke; Equilibrium; Evans blue stain; Extravasation; Feasibility Studies; Functional disorder; Gene Expression Profiling; Genes; Goals; HIV; HIV Envelope Protein gp120; HIV-1; Health; Highly Active Antiretroviral Therapy; Homing; In Vitro; Incidence; Infection; Injury; Ions; Legal; Link; MCAM gene; Malignant Neoplasms; Marijuana; Methamphetamine; Microfluidics; Modeling; Molecular; Molecular Biology Techniques; Mus; Neurosciences Research; Nicotine; Opiates; Pathogenesis; Pathway interactions; Permeability; Pharmaceutical Preparations; Phenotype; Pilot Projects; Population; Proteins; Reporting; Serum; Smoker; Stem cells; Testing; Tobacco; Tobacco use; UCHL1 gene; Virulence Factors; Work; base; bone; genome-wide; in vivo; magnetic beads; microbial; molecular marker; nicotine abuse; novel; outcome forecast; pathogen; peripheral blood; pre-clinical; repaired; response; tool; transcriptome sequencing

DESCRIPTION (provided by applicant): The lack of specific cellular markers for quantitative assessment of pathogenic damage to the blood-brain barrier (BBB), which is mainly constituted by microvascular endothelial cells (BMEC), is one of the most challenging issues in central nervous system (CNS) disorders induced by various pathogenic insults, including AIDS, and the use of illegal drugs [e.g., methamphetamine (METH), marijuana, opiates] and legal substances such as alcohol and nicotine (NT), a major component of environmental tobacco smoke. The incidence of NeuroAIDS is higher or accelerated among drug users and tobacco smokers. Alterations of brain microvasculature and the disruption of the BBB integrity are commonly associated with the use of drugs and HIV-1 infection in the era of HAART. In our previous work it was found that dysfunction of BMEC could be induced by NT, meningitic pathogens and microbial factors, including HIV-1 virulence factors (VFs) gp41 and gp120. The overall goal of this pilot project is to perform feasibility studies on the development of new cell-based biomarkers of the BBB disorders induced by drug abuse (NT) and HIV VFs (gp120 and gp41) through a combination of in vitro(BMEC)/in vivo (mouse) BBB models and high throughput single-cell profiling approaches. Our studies have shown that mice treated with NT and gp120 resulted in a synergistic increase in blood levels of circulating BMEC (cBMEC) [UEA+(EC marker)/CD146+(EC marker)/S100B+ (brain marker)] as well as endothelial progenitor cells (EPC)[UEA+/CD146+/CD133+(progenitor cell marker)]. NT and gp120 were able to significantly increase the serum levels of UCHL1 (a new BBB marker) as well as S100B in mice, which are correlated with the changes in cBMEC and EPC. Based on these findings, we have hypothesized that cBMEC in the peripheral blood, which are endowed with a full-blown BBB phenotype, are dynamically shed from the BBB. The replacement of shed or dead BMEC could occur through division of surrounding EC and homing of EPC, which are derived from bone marrow. The integrity of the BBB is associated with the functional changes in the brain microvasculatures and maintained by the balance between BMEC shedding/death rate and cell renewal rate. Peripheral blood cBMEC detected by molecular and single-cell profiling approaches can be used as a novel genomewide, cell-based multiple biomarker and gene network detection tool for the BBB injury, which is induced by NT, gp120 and other pathogenic insults. The availability of cBMEC as peripheral blood biomarkers in the CNS diseases will have a great impact in neuroscience research on drug abuse and neuroAIDS. Our hypotheses will be tested with the preclinical proof-of-concept studies through the following two Specific Aims: (1). Establish in vitro/in vivo experimental approaches for the characterization of cBMEC and UCHL1 as novel biomarkers of BBB injury induced by NT and HIV-1 gp120. (2). Determine the correlation between disease-specific BBB injury and blood levels of biomarkers by molecular typing and single-cell profiling of cBMEC.

描述（申请人提供）：血脑屏障（BBB）主要由微血管内皮细胞（BMEC）构成，缺乏定量评估致病性损伤的特异性细胞标志物，是中枢神经系统中最具挑战性的问题之一。由各种病原体损伤（包括艾滋病）以及使用非法药物 [例如甲基苯丙胺 (METH)、大麻、阿片类药物] 和酒精等合法物质引起的神经系统 (CNS) 疾病和尼古丁（NT），环境烟草烟雾的主要成分。吸毒者和吸烟者中神经艾滋病的发病率更高或更快。 HAART 时代，脑微血管系统的改变和 BBB 完整性的破坏通常与药物的使用和 HIV-1 感染有关。我们之前的工作发现，NT、脑膜炎病原体和微生物因素（包括 HIV-1 毒力因子（VFs）gp41 和 gp120）可诱导 BMEC 功能障碍。该试点项目的总体目标是通过体外（BMEC）/药物滥用（NT）和HIV VF（gp120和gp41）相结合，开发新的基于细胞的BBB疾病生物标志物的可行性研究体内（小鼠）BBB 模型和高通量单细胞分析方法。我们的研究表明，用 NT 和 gp120 治疗的小鼠导致循环 BMEC (cBMEC) [UEA+（EC 标记物）/CD146+（EC 标记物）/S100B+（脑标记物）] 以及内皮祖细胞的血液水平协同增加(EPC)[UEA+/CD146+/CD133+（祖细胞标记）]。 NT和gp120能够显着增加小鼠血清中UCHL1（一种新的BBB标记物）和S100B的水平，这与cBMEC和EPC的变化相关。基于这些发现，我们假设外周血中的 cBMEC 具有成熟的 BBB 表型，是从 BBB 动态脱落的。脱落或死亡的 BMEC 的更换可以通过周围 EC 的分裂和源自骨髓的 EPC 归巢来实现。 BBB 的完整性与大脑微血管的功能变化相关，并通过 BMEC 脱落/死亡率和细胞更新率之间的平衡来维持。通过分子和单细胞分析方法检测的外周血 cBMEC 可用作新型全基因组、基于细胞的多重生物标志物和基因网络检测工具，用于检测由 NT、gp120 和其他致病性损伤引起的 BBB 损伤。 cBMEC 作为中枢神经系统疾病的外周血生物标志物的可用性将对药物滥用和神经艾滋病的神经科学研究产生巨大影响。我们的假设将通过以下两个具体目标通过临床前概念验证研究进行测试：（1）。建立体外/体内实验方法来表征 cBMEC 和 UCHL1 作为 NT 和 HIV-1 gp120 诱导的 BBB 损伤的新型生物标志物。 （2）。通过 cBMEC 的分子分型和单细胞分析确定疾病特异性 BBB 损伤与生物标志物血液水平之间的相关性。