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: 8604150
• 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/8604150
• 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; public health relevance; repaired; response; tool; transcriptome sequencing; 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; public health relevance; 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.

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.,甲基苯丙胺（甲基苯丙胺），大麻，阿片类药物]和诸如酒精和尼古丁（NT）之类的合法物质，这是环境烟草烟雾的主要组成部分。在吸毒者和吸烟者中，神经助剂的发病率更高或加速。在HAART时代，脑微举行的改变和BBB完整性的破坏通常与药物和HIV-1感染的使用有关。在我们以前的工作中，发现BMEC的功能障碍可以由NT，脑膜炎病原体和微生物因子（包括HIV-1毒力因子（VFS）GP41和GP120）诱导。该试点项目的总体目标是对由药物滥用（NT）和HIV VFS（GP120和GP41）引起的新的基于细胞的BBB疾病的生物标志物进行可行性研究，这是通过体外（BMEC）/In vivo（BMEC）/In vivo（Mouse）BBB模型和高遍历单孔型单细胞素材的组合。我们的研究表明，用NT和GP120处理的小鼠导致循环BMEC（CBMEC）[UEA+（EC Marker）/CD146+（EC Marker）+（EC Marker）/S100B+（Brain Marker）的血液水平有协同增加，并且是内皮阶梯细胞（EPC）[UEA+/CD114666333333333333333。 NT和GP120能够显着增加与CBMEC和EPC的变化相关的小鼠中UCHL1（新的BBB标记）以及S100B的血清水平。基于这些发现，我们假设外周血中的CBMEC被赋予了成熟的BBB表型，它是从BBB动态脱落的。可以通过分裂周围的EC和EPC的归巢（源自骨髓）来替代棚屋或死亡BMEC。 BBB的完整性与大脑微脉管发生的功能变化有关，并通过BMEC脱落/死亡率和细胞更新率之间的平衡维持。通过分子和单细胞分析方法检测到的外周血CBMEC可以用作BBB损伤的新型基因组，基于细胞的多个生物标志物和基因网络检测工具，该工具由NT，GP120和其他致病性损伤引起。 CBMEC作为中枢神经系统疾病中外周血生物标志物的可用性将对药物滥用和神经助剂的神经科学研究产生重大影响。我们的假设将通过以下两个具体目的进行临床前概念验证研究测试：（1）。建立体外/体内实验方法，以表征CBMEC和UCHL1为NT和HIV-1 GP120诱导的BBB损伤的新生物标志物。 （2）。通过分子键入和CBMEC的单细胞分析，确定疾病特异性BBB损伤与血液水平之间的相关性。