Lymphatic Vessel Imaging

淋巴管成像

• 批准号: 7772428
• 负责人: Nancy H. Ruddle
• 金额: $ 24.83万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-01-15 至 2011-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7772428
• 关键词: Abbreviations; Antigen-Presenting Cells; Antigens; Autoimmunity; Automobile Driving; Blood Circulation; CD44 Antigens; Cells; Characteristics; Chronic; Chylothorax; Communicable Diseases; Development; Endothelial Cells; Exhibits; Functional disorder; Galactosidase; Genes; Goals; Greater sac of peritoneum; Green Fluorescent Proteins; Growth; High Endothelial Venule; Homeobox; Image; Imaging Device; Immune response; Immune system; Immunization; Inflammation; Insulin; Intercellular Fluid; LacZ Genes; Life; Liquid substance; Lymph; Lymphangiogenesis; Lymphatic; Lymphatic vessel; Lymphedema; Lymphoid; Lymphoid Tissue; Magnetic Resonance Imaging; Malignant Neoplasms; Microscopy; Mus; Nose; Organ; PNAd; Pancreas; Peripheral; Process; Proteins; Rattus; Regulation; Reporter; Reporter Genes; Site; Staining method; Stains; TNF gene; Techniques; Technology; Temperature; Therapeutic; Time; Tissues; Transgenes; Transgenic Mice; Transplantation; Tumor Necrosis Factor-Beta; Tumor Necrosis Factor-alpha; Tumor Necrosis Factors; Vascular System; in vivo; injured; insight; interstitial; lipid transport; lymph nodes; periodate-lysine-paraformaldehyde; podoplanin; promoter; public health relevance; red fluorescent protein; residence; sulfotransferase; theories

DESCRIPTION (provided by applicant): The long term goal of this R21 project is to develop tools for imaging lymphatic vessels (LVs) in living mice. Lymphatic vessels drain interstitial fluid by means of valve-like openings, they transport lipids and they participate in the immune system. They transport antigen and antigen presenting cells to lymph nodes and cells from the lymph node to the circulation. When lymphatic vessels are disrupted, lymphedema, the accumulation of lymph in the interstitial tissues, occurs. Immunological functions are thwarted. Lymphangiogenesis occurs in inflammation at the site of immunization and in lymph nodes (secondary lymphoid organs) where there is an interaction between LVs and high endothelial venules (HEVs). Lymphangiogenesis occurs in chronic inflammation in lymphoid accumulations in ectopic sites, called tertiary lymphoid tissues (TLOs) that have many characteristics of lymph nodes. Mice transgenic for the rat insulin promoter (RIP) driving lymphotoxin-1 (RIPLT1) exhibit such TLOs in the pancreas. Techniques that have been used to develop mice whose HEVs can be imaged via green fluorescent protein will be used here to develop mice whose LVs can be similarly imaged via a red fluorescent protein. To attain this goal, the following specific aims will be accomplished: 1. To develop transgenic mice whose lymphatic vessels express fluorescent proteins. The pCLASPER recombineering technology will be used to isolate LV genes (prox1, lyve-1, or podoplanin), insert red fluorescent reporter genes, such as tdTomato, and produce transgenic mice. 2. To evaluate lymphatic reporter transgenic mice for fidelity of expression. Mice transgenic for lymphatic vessel reporter constructs will be analyzed for co- expression of endogenous LV genes and transgenes in several tissues in the steady state, in lymph nodes after immunization, and in TLOs in RIPLT1 mice. 3. To evaluate LVs in living mice. Multiphoton in vivo microscopy will be used to evaluate fluorescent LVs. Interactions between (green) high endothelial venules and (red) lymphatic vessels in the lymph nodes of immunized mice will be analyzed in real time. A dynamic understanding of lymphangiogenesis and LV function will provide therapeutic insight into lymphatic function and dysfunction in lymphedema. The development of imaging tools for lymphatic vessels will provide insight into how LVs and HEVs are regulated, maintained, and cooperate during an immune response and provide insight into lymphangiogenesis in inflammation, transplantation, autoimmunity, infectious diseases, and cancer. PUBLIC HEALTH RELEVANCE: A thorough understanding of lymphangiogenesis and lymphatic vessel function will provide therapeutic insight into lymphatic function and dysfunction in conditions such as lymphedema. Imaging tools for lymphatic vessels will provide insight into how this vascular system is maintained and regulated during an immune response and provide insight into lymphangiogenesis in inflammation, transplantation, autoimmunity, infectious diseases, and cancer. )

描述（由申请人提供）：该R21项目的长期目标是开发用于活体小鼠淋巴管（LV）成像的工具。淋巴管通过瓣膜样开口排出间质液，它们运输脂质并参与免疫系统。它们将抗原和抗原呈递细胞转运到淋巴结，并将细胞从淋巴结转运到循环。当淋巴管被破坏时，会发生水肿，即淋巴液在间质组织中的积聚。免疫功能受到阻碍。淋巴管生成发生在免疫部位和淋巴结（次级淋巴器官）的炎症中，其中LV和高内皮微静脉（HEV）之间存在相互作用。淋巴管生成发生在异位部位的淋巴积聚中的慢性炎症中，称为三级淋巴组织（TLO），其具有淋巴结的许多特征。对于大鼠胰岛素启动子（RIP）驱动的胰高血糖素-1（RIPLT 1）转基因的小鼠在胰腺中表现出这样的TLO。已经用于开发其HEV可以通过绿色荧光蛋白成像的小鼠的技术将在这里用于开发其LV可以通过红色荧光蛋白类似地成像的小鼠。为实现这一目标，将实现以下具体目标：1.建立淋巴管表达荧光蛋白的转基因小鼠。pCLASPER重组工程技术将用于分离LV基因（pCL 1、lyve-1或podoplanin），插入红色荧光报告基因（如tdTomato），并产生转基因小鼠。2.评价淋巴报告基因转基因小鼠的表达保真度。将分析淋巴管报告基因构建体转基因小鼠在稳态下的几种组织中、免疫后的淋巴结中和RIPLT 1小鼠的TLO中内源性LV基因和转基因的共表达。3.评价活体小鼠的LV。多光子体内显微镜将用于评价荧光LV。将真实的分析免疫小鼠淋巴结中（绿色）高内皮微静脉和（红色）淋巴管之间的相互作用。对淋巴管生成和左室功能的动态了解将为水肿中的淋巴功能和功能障碍提供治疗见解。淋巴管成像工具的开发将提供对LV和HEV在免疫应答期间如何调节、维持和合作的深入了解，并提供对炎症、移植、自身免疫、感染性疾病和癌症中淋巴管生成的深入了解。 公共卫生相关性：对淋巴管生成和淋巴管功能的透彻理解将提供对诸如水肿等病症中的淋巴功能和功能障碍的治疗见解。淋巴管的成像工具将提供对免疫反应期间如何维持和调节该血管系统的深入了解，并提供对炎症，移植，自身免疫，感染性疾病和癌症中淋巴管生成的深入了解。)