Progenitor Cells for High Endothelium in the Immune Response

免疫反应中高内皮的祖细胞

• 批准号: 9755349
• 负责人: EUGENE C BUTCHER
• 金额: $ 50.97万
• 依托单位: PALO ALTO VETERANS INSTIT FOR RESEARCH
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-25 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9755349
• 关键词: Antibodies; Antigens; Area; Atherosclerosis; Autoimmune Diseases; Autoimmune Process; Biology; Blood; Blood Vessels; Blood capillaries; Capillary Endothelial Cell; Cells; Chronic; Clone Cells; Cytometry; Development; Disease; Down-Regulation; Endothelial Cells; Endothelium; Enlargement of lymph nodes; Enterobacteria phage P1 Cre recombinase; Flow Cytometry; Gene Expression Profiling; Gene Targeting; Generations; Genetic; Goals; Gut associated lymphoid tissue; High Endothelial Venule; Histology; Homeostasis; Image; Immune; Immune response; Immunity; Immunization; Immunize; Immunofluorescence Immunologic; Immunologics; Immunology; Inflammation; Inflammatory; Intestines; Investigation; Kinetics; Label; Lead; Leukocytes; Location; Lymphocyte; Lymphoid; Lymphoid Tissue; Malignant Neoplasms; Methods; Molecular; Monitor; Mus; Outcome; Pathogenicity; Pathologic; Pathology; Pathway interactions; Phenotype; Physiological; Population; Proliferating; Reporter; Rest; Role; Signal Transduction; Site; Skin; Specificity; Stem cells; Surface Antigens; System; Testing; Therapeutic; Time; Tissue imaging; Tissues; Up-Regulation; Vaccination; angiogenesis; base; high dimensionality; immunoregulation; in vivo; innovation; insight; lymph nodes; migration; notch protein; novel; novel strategies; novel therapeutic intervention; postcapillary venule; progenitor; programs; recruit; response; single cell analysis; stem; stem cell population; stem-like cell; tool; trafficking; transcriptomics; tumor

PROJECT SUMMARY / ABSTRACT High endothelial venules (HEV) are specialized post capillary portals for lymphocyte entry into lymphoid tissues and sites of chronic inflammation from the blood. They regulate immune cell trafficking in physiologic and pathologic settings including autoimmune diseases and cancer. HEV in lymph nodes draining sites of immune challenge proliferate extensively to support enhanced lymphocyte recruitment, but the endothelial precursors that give rise to high endothelium and the molecular pathways that control their proliferation and differentiation are unclear. Our transcriptomic and immunologic studies reveal that capillary endothelial cells (CapEC) express multiple markers classically associated with stem and progenitor cells, leading to the hypothesis that capillary phenotype EC comprise a population of stem cell-like blood endothelial progenitors that contribute to HEV expansion during the immune response. Under Aim 1 we will apply state-of-the-art single cell immunofluorescence and high dimensional mass label (CyTOF) flow cytometry to uncover the diversity of EC subsets in LN and in extra lymphoid sites of immune cell recruitment, define the kinetics and subset-specificity of their proliferative responses to immune challenge, and reveal the time course of emergence of transitional phenotypes leading to the amplification and maturation of functional HEV. Trajectory analyses will reveal developmental relationships of identified subsets including identified progenitors, and immunofluorescence histology and confocal tissue imaging will define their location within the vasculature. In Aim 2, innovative fate mapping approaches will elucidate precursor-product relationships among BEC subsets and will define clonal contributions of precursors to capillary and high endothelium. Aim 3 will apply transcriptional profiling of induced EC subsets in combination with pan-EC and novel capillary EC-specific inducible gene targeting systems to define molecular mechanisms of HEV homeostasis and pathways regulating progenitor cell activation, amplification and contribution to HEV expansion in the immune response. Comprehensive phenotypic analyses of blood endothelial cell subsets and their responses to immunization will open up new areas of investigation in vascular biology and immunology. Elucidation of the mechanisms of endothelial cell specialization and homeostasis, including mechanisms regulating endothelial cells that control lymphocyte traffic, will lead to novel targets and approaches for the control of autoimmune inflammation and for therapeutic regulation of immune cell traffic for vaccination and cancer immunity.

项目概要/摘要 高内皮微静脉 (HEV) 是淋巴细胞进入淋巴组织的特殊毛细血管后入口 以及血液中慢性炎症的部位。它们调节生理和免疫细胞的运输 病理环境，包括自身免疫性疾病和癌症。淋巴结引流免疫部位的 HEV 挑战广泛增殖以支持增强的淋巴细胞募集，但内皮前体细胞 产生高内皮细胞和控制其增殖和分化的分子途径 不清楚。我们的转录组学和免疫学研究表明，毛细血管内皮细胞 (CapEC) 表达 多个标记物经典地与干细胞和祖细胞相关，导致毛细血管的假设 表型 EC 包含一群有助于 HEV 的干细胞样血液内皮祖细胞 免疫反应期间的扩张。在目标 1 下，我们将应用最先进的单细胞 免疫荧光和高维质量标签 (CyTOF) 流式细胞术揭示 EC 的多样性 淋巴结和免疫细胞募集的淋巴外位点的子集，定义了动力学和子集特异性 它们对免疫挑战的增殖反应，并揭示过渡性细胞出现的时间过程 表型导致功能性 HEV 的扩增和成熟。轨迹分析将揭示 已识别亚群的发育关系，包括已识别祖细胞和免疫荧光 组织学和共聚焦组织成像将确定它们在脉管系统内的位置。目标2，创新命运 作图方法将阐明 BEC 子集之间的前体-产物关系，并定义克隆 前体对毛细血管和高内皮细胞的贡献。目标 3 将应用诱导的转录分析 EC 子集与泛 EC 和新型毛细血管 EC 特异性诱导基因靶向系统相结合 定义 HEV 稳态的分子机制和调节祖细胞激活的途径， 免疫反应中 HEV 扩增的放大和贡献。 血液内皮细胞亚群及其对免疫反应的综合表型分析将 开辟血管生物学和免疫学的新研究领域。阐明其机制 内皮细胞特化和稳态，包括控制内皮细胞的调节机制 淋巴细胞运输，将带来控制自身免疫炎症和治疗的新靶点和方法 用于疫苗接种和癌症免疫的免疫细胞运输的治疗调节。