Lymphatic Vessel Imaging

淋巴管成像

基本信息

批准号：
8013023
负责人：
Nancy H. Ruddle
金额：
$ 20.69万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
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/8013023
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 Lymphocyte 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 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

项目摘要

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活小鼠中成像淋巴管（LVS）的工具。淋巴管通过类似瓣膜的开口来排出间质液，它们运输脂质并参与免疫系统。他们将抗原和抗原呈递细胞从淋巴结和循环中传递到淋巴结和细胞。当淋巴血管破坏时，会发生淋巴水肿，淋巴在间质组织中的积累。免疫功能受到挫败。淋巴管生成发生在免疫部位和淋巴结（继发性淋巴机构）的炎症中，其中LVS与高内皮静脉（HEVS）之间存在相互作用。淋巴管生成发生在异位部位的淋巴结累积中，称为淋巴结特征，称为第三纪淋巴组织（TLOS）。大鼠胰岛素启动子（RIP）驱动淋巴毒素1（RIPLT1）的小鼠转基因在胰腺中表现出这种TLO。在这里将使用用于开发其HEV的小鼠的技术，将在这里使用绿色荧光蛋白成像，以开发小鼠，其LV可以通过红色荧光蛋白进行类似成像。为了实现这一目标，将实现以下特定目标：1。开发淋巴管表达荧光蛋白的转基因小鼠。 PCLASPER重新组合技术将用于分离LV基因（Prox1，Lyve-1或Podoplanin），插入红色荧光报告基因，例如TDTOMATO，并产生转基因小鼠。 2。评估淋巴报告转基因小鼠的表达保真度。将分析用于淋巴血管报告基因构建体的小鼠转基因，以在稳态的几个组织中，免疫后的淋巴结中的几个组织中的内源性LV基因和转基因共同表达，在RIPLT1小鼠中的TLOS中。 3。评估活小鼠的LV。多光体在体内显微镜将用于评估荧光LV。（绿色）高内皮细胞和（红色）免疫小鼠淋巴结（红色）淋巴管之间的相互作用将进行实时分析。对淋巴管生成和LV功能的动态理解将为淋巴水肿的淋巴功能和功能障碍提供治疗见解。淋巴管的成像工具的开发将提供有关在免疫反应期间如何调节，维持和合作的LV和HEV的洞察力，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见解。公共卫生相关性：对淋巴管生成和淋巴血管功能的透彻理解将为淋巴水肿等淋巴功能和功能障碍提供治疗性见解。淋巴管的成像工具将提供有关在免疫反应期间如何维持和调节该血管系统的洞察力，并洞悉炎症，移植，自身免疫，感染性疾病和癌症中淋巴管生成。）