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在活体小鼠身上对淋巴管(LV)进行成像的工具。淋巴管通过瓣膜状的开口排出间质液体，它们运输脂质，并参与免疫系统。它们将抗原和抗原提呈细胞运送到淋巴结，并将细胞从淋巴结运送到循环中。当淋巴管被破坏时，就会发生淋巴水肿，即淋巴在间质组织中的聚集。免疫功能受挫。淋巴管生成发生在免疫部位的炎症和淋巴结(次级淋巴器官)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淋巴聚集的慢性炎症中，称为三级淋巴组织(TLO)，具有许多淋巴结节的特征。转大鼠胰岛素启动子(RIP)驱动淋巴毒素-1(RIPLT1)的小鼠在胰腺中表现出这样的TLO。已经被用来培育其HEV可以通过绿色荧光蛋白成像的小鼠的技术将在这里被用于开发其左肺可以类似地通过红色荧光蛋白成像的小鼠。为了实现这一目标，将完成以下具体目标：1.建立淋巴管表达荧光蛋白的转基因小鼠。PCLASPER重组工程技术将被用于分离LV基因(PROX1，Lyve-1，或podoplan in)，插入红色荧光报告基因，如tdTomato，并产生转基因小鼠。2.检测淋巴道报告基因转基因小鼠的表达保真度。转基因淋巴管报告基因的小鼠将分析内源性LV基因和外源基因在稳定状态下的几个组织中、免疫后的淋巴结和RIPLT1小鼠的TLO中的共同表达。3.评价活体小鼠的左肺功能。多光子活体显微镜将被用来评估荧光LV。将实时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。对淋巴管生成和左心室功能的动态了解将为淋巴水肿的淋巴功能和功能障碍提供治疗性的见解。淋巴管成像工具的开发将有助于深入了解免疫反应期间LV和HEV是如何调节、维持和协作的，并有助于深入了解炎症、移植、自身免疫、传染病和癌症中的淋巴管生成。 公共卫生相关性：彻底了解淋巴管生成和淋巴管功能将提供淋巴功能和淋巴水肿等情况下的功能障碍的治疗洞察力。淋巴管成像工具将提供对免疫反应期间血管系统如何维持和调节的洞察，并提供对炎症、移植、自身免疫、传染病和癌症中淋巴管生成的洞察。)