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.

项目摘要 /摘要 高内皮静脉（HEV）是专门的毛细血管门户，用于淋巴细胞进入淋巴组织 以及血液中慢性炎症的部位。他们调节生理学和 病理环境，包括自身免疫性疾病和癌症。免疫的淋巴结排水位点的HEV 挑战广泛的挑战以支持增强的淋巴细胞募集，但内皮前体 这会引起高内皮以及控制其增殖和分化的分子途径 不清楚。我们的转录组和免疫学研究表明，毛细管内皮细胞（CAPEC）表达 与茎和祖细胞经典相关的多个标记，导致假设是毛细管 表型EC包括有助于HEV的干细胞样血液内皮祖细胞的种群 免疫反应期间的扩张。在AIM 1下，我们将应用最先进的单个单元格 免疫荧光和高维质量标签（Cytof）流式细胞仪，以发现EC的多样性 LN和免疫细胞募集的额外淋巴部位的子集，定义动力学和子集特异性 他们对免疫挑战的增生反应，并揭示了过渡的时间过程 表型导致功能HEV的扩增和成熟。轨迹分析将揭示 已确定子集的发展关系，包括已识别的祖细胞和免疫荧光 组织学和共聚焦组织成像将定义其在脉管系统中的位置。在AIM 2中，创新的命运 映射方法将阐明BEC子集之间的前体产物关系，并将定义克隆 前体对毛细血管和高内皮的贡献。 AIM 3将应用诱导的转录分析 与PAN-EC和新型毛细管EC特异性诱导基因靶向系统结合 定义HEV稳态和调节祖细胞激活的途径的分子机制， 免疫反应中HEV扩展的扩增和贡献。 血液内皮细胞亚群的全面表型分析及其对免疫的反应将 开设血管生物学和免疫学研究的新研究领域。阐明机制 内皮细胞专业化和稳态，包括调节控制内皮细胞的机制 淋巴细胞流量将导致自身免疫性炎症控制和用于控制的新目标和方法 免疫细胞流量的治疗调节，以进行疫苗接种和癌症免疫。