Transcriptional Profiling of Human High Endothelial Venules

人类高内皮小静脉的转录谱

• 批准号: 9212639
• 负责人: EUGENE C BUTCHER
• 金额: --
• 依托单位: VETERANS ADMIN PALO ALTO HEALTH CARE SYS
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2020-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9212639
• 关键词: ATAC-seq; Adenoidal structure; Adhesions; Affect; Alzheimer' s Disease; Animal Model; Animals; Antibodies; Area; Arthritis; Autoimmune Process; Autopsy; Binding; Biology; Blood; Blood Vessels; Blood capillaries; CCL21 gene; Capillary Endothelial Cell; Cardiovascular Diseases; Cardiovascular system; Cell Communication; Cell Separation; Cells; Chromatin; Chronic; Clinical; Data; Diabetes Mellitus; Disease; Dissociation; Distal; Drug Targeting; Endothelial Cells; Endothelium; Enhancers; Enzymes; Epithelial; Gene Expression; Gene Expression Regulation; General Population; Genes; Genetic Transcription; Goals; Gut associated lymphoid tissue; High Endothelial Venule; Homing; Human; Immune; Immune response; Immunology; Inflammation; Inflammation Process; Inflammatory; Inflammatory Bowel Diseases; Inflammatory Response; Investigation; L-Selectin; Lead; Leukocyte Trafficking; Leukocytes; Literature; Lymphocyte; Lymphoid Tissue; Maps; Mediating; Molecular; Molecular Profiling; Mus; Ontology; Operative Surgical Procedures; Organ; Organ Specificity; Outcome; PECAM1 gene; PNAd; Pathologic; Pathway interactions; Pattern; Peripheral; Protocols documentation; RNA; Recruitment Activity; Resources; Signal Pathway; Site; Specimen; Surface; Tissue Procurements; Tissues; Tonsil; Translating; Translations; Vascular Endothelium; Veterans; adhesion receptor; candidate marker; capillary; chemokine; epigenomics; genetic signature; insight; lymph nodes; man; novel; postcapillary venule; programs; promoter; public health relevance; receptor; selective expression; species difference; targeted treatment; therapeutic target; trafficking; transcription factor; transcriptome; transcriptomics; vascular addressins; whole genome

 DESCRIPTION (provided by applicant): Our goal is to elucidate the transcriptional specialization of human high endothelial venules (HEV), unique post-capillary venules that control lymphocyte homing and thus immune and inflammatory responses. Most studies of HEV, and the only comprehensive studies of gene expression by HEV, have been carried out in mice: relatively little is known about human HEV or indeed, about the gene programs underlying the segmental and tissue specialization of blood endothelial cells in man. Notably, several vascular molecules implicated in lymphocyte homing in the mouse, including the peripheral lymph node "addressin", VAP1, CCL21 and others, are regulated differently in humans. We recently completed comprehensive transcriptomic studies of mouse lymphoid tissue high endothelial cells (HEC) and enriched capillary EC (CAP). Here we shall apply our established protocols to perform whole genome expression analyses of human lymphoid tissue HEV and CAP. Aims include: 1) To perform whole genome expression profiling of high endothelial cells (HEC) and capillary endothelial cells (CAP) from human tonsils, adenoids, peripheral lymph nodes (PLN), and appendix. We will apply existing protocols for endothelial dissociation from fresh human lymphoid tissues, and will purify HEC and capillary EC using FACS cell sorting protocols with established HEC and pan-EC markers. RNA will be submitted for whole genome expression profiling. 2) To define the segmental (HEV vs CAP) and tissue-specific (tonsil or adenoid, PLN, appendix) transcriptional specialization of human high endothelial cells (HEC). We shall elucidate vascular programs that define HEC specialization (and thus lymphocyte homing) by comparing the transcriptomes of HEC and of capillary endothelium (CAP), which unlike HEC inhibit leukocyte adhesion. We will also identify gene sets expressed in HEC in a tissue-specific fashion, genes likely to encode mechanisms that control vascular addressins or chemo attractants for tissue-specific lymphocyte recruitment. 3) To compare and contrast human vs. mouse HEV-, CAP- and tissue-specific HEC gene signatures, and to identify evolutionarily conserved transcriptional programs of HEC. We shall define differences in HEC gene expression in humans vs. mice, the most studied animal model. We will identify genes with conserved patterns of HEC vs CAP, or of tissue-specific HEC expression: such genes are likely to encode molecular mechanisms (transcription factors, adhesion or attractant molecules, signaling pathways) critical to HEV function. Comprehensive analyses of human HEV and capillary EC transcriptomes will open up new areas of investigation in vascular biology and immunology, and will inform efforts to translate mouse studies to the clinical arena. The data generated will constitute a valuable resource for elucidation of mechanisms of vascular control of lymphocyte recruitment, and may lead to novel targets and approaches for the control of autoimmune and other pathologic inflammatory disorders.

 描述(由申请人提供)： 我们的目标是阐明人类高内皮微静脉(HEV)的转录特化，这是一种独特的毛细血管后微静脉，控制淋巴细胞归巢，从而控制免疫和炎症反应。大多数关于HEV的研究，也是唯一对HEV基因表达的全面研究，都是在小鼠身上进行的：对人类HEV，或者实际上，关于人类血液内皮细胞节段性和组织特化的基因程序知之甚少。值得注意的是，几种与小鼠淋巴细胞归巢有关的血管分子，包括外周淋巴结“Addressin”、VAP1、CCL21和其他分子，在人类中受到不同的调控。我们最近完成了对小鼠淋巴组织高内皮细胞(HEC)和富集型毛细血管内皮细胞(CAP)的全面转录研究。在这里，我们将应用我们建立的方案来执行人类淋巴组织HEV和CAP的全基因组表达分析。目的包括：1)对人扁桃体、腺样体、周围淋巴结和阑尾的高内皮细胞(HEC)和毛细血管内皮细胞(CAP)进行全基因组表达谱分析。我们将应用现有的从新鲜人类淋巴组织中分离内皮细胞的方案，并将使用已建立的HEC和PAN-EC标记的FACS细胞分选方案来纯化HEC和毛细血管EC。RNA将被提交用于全基因组表达谱分析。2)明确人高内皮细胞(HEC)的节段性(HEV与CAP)和组织特异性(扁桃体或腺样体、PLN、阑尾)转录的特异化。我们将通过比较HEC和毛细血管内皮细胞(CAP)的转录本来阐明定义HEC特化(从而定义淋巴细胞归巢)的血管程序，与HEC不同，毛细血管内皮细胞抑制白细胞黏附。我们还将确定在HEC中以组织特异性方式表达的基因集，这些基因可能编码控制组织特异性淋巴细胞募集的血管地址蛋白或化学诱导剂的机制。3)比较人和小鼠HEV、CAP和组织特异性HEC基因特征，并鉴定HEC在进化上保守的转录程序。我们将确定人类和小鼠HEC基因表达的差异，小鼠是研究最多的动物模型。我们将确定具有HEC与CAP保守模式的基因，或组织特异性HEC表达的基因：这些基因可能编码对HEV功能至关重要的分子机制(转录因子、黏附或吸引分子、信号通路)。对人类HEV和毛细血管EC转录本的全面分析将开辟血管生物学和免疫学研究的新领域，并将为将小鼠研究转化为临床领域的努力提供信息。所产生的数据将成为阐明淋巴细胞募集的血管控制机制的宝贵资源，并可能导致控制自身免疫和其他病理性炎症性疾病的新靶点和方法。