Transcriptional Profiling of Human High Endothelial Venules

人类高内皮小静脉的转录谱

基本信息

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
项目类别：
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&apos 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

项目摘要

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静脉（HEV）的转录专业化，独特的后毛细血管静脉，以控制淋巴细胞的归纳，从而免疫学和炎症反应。大多数HEV的研究以及HEV对基因表达的唯一全面研究是在小鼠中进行的：关于人HEV或实际上，关于人类HEV或实际上，对人类血液内皮细胞的分段和组织专业化的基因程序知之甚少。值得注意的是，在小鼠中淋巴细胞归巢中实施的几种血管分子，包括外周淋巴结“地址”，VAP1，CCL21等，在人类中受到不同的调节。我们最近完成了小鼠淋巴组织高内皮细胞（HEC）和富含毛细血管EC（CAP）的全面转录组研究。在这里，我们将应用我们既定的方案来执行人淋巴组织HEV和CAP的整个基因组表达分析。目的包括：1）从人扁桃体，腺样体，外周淋巴结（PLN）和附录进行高内皮细胞（HEC）和毛细血管内皮细胞（CAP）的全基因组表达分析。我们将使用现有的方案从新鲜的人淋巴组织中进行内皮解离，并使用具有既定HEC和PAN-EC标记的FACS细胞分类方案纯化HEC和毛细管EC。 RNA将提交全基因组表达分析。 2）定义人类高内皮细胞（HEC）的分段（HEV与CAP）和组织特异性（扁桃体或腺样体，PLN，附录）的转录专业化（HEC）。我们将通过比较HEC和毛细管内皮（CAP）的转录组来阐明定义HEC专业化（以及淋巴细胞归巢）的血管计划，这些毛细血管内皮（CAP）与HEC抑制白细胞粘附不同。我们还将以组织特异性的方式鉴定在HEC中表达的基因集，这些基因可能编码控制血管地址的机制或用于组织特异性淋巴细胞募集的化学吸引剂。 3）要比较和对比人类与小鼠HEV，CAP和组织特异性HEC基因的特征，并确定HEC的进化配置的转录程序。我们将定义人类中HEC基因表达的差异与研究最多的动物模型。我们将鉴定具有HEC与CAP的组成模式或组织特异性HEC表达的基因：此类基因可能编码对HEV功能至关重要的分子机制（转录因子，粘合剂或有吸引力的分子，信号通路）。对人HEV和毛细血管EC转录组的全面分析将为血管生物学和免疫学投资的新领域，并将为将小鼠研究转化为临床领域的努力。生成的数据将构成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性和其他病理炎症性疾病的新目标和方法。