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用于活体小鼠淋巴管 (LV) 成像的工具。淋巴管通过瓣膜状开口排出间质液，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至淋巴结，并将细胞从淋巴结转运至循环系统。当淋巴管受到破坏时，就会发生淋巴水肿（淋巴液在间质组织中积聚）。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淋巴积聚的慢性炎症中，称为三级淋巴组织（TLO），具有淋巴结的许多特征。驱动淋巴毒素 1 (RIPLT1) 的大鼠胰岛素启动子 (RIP) 转基因小鼠在胰腺中表现出此类 TLO。已用于开发可通过绿色荧光蛋白对 HEV 进行成像的小鼠的技术将在这里用于开发可通过红色荧光蛋白对 LV 进行类似成像的小鼠。为实现这一目标，将完成以下具体目标： 1.培育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（prox1、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.评估淋巴报告基因转基因小鼠的表达保真度。将分析淋巴管报道基因构建体转基因小鼠在稳定状态下的多个组织中、免疫后的淋巴结中以及RIPLT1小鼠的TLO中的内源LV基因和转基因的共表达。 3. 评估活体小鼠的 LV。多光子体内显微镜将用于评估荧光 LV。将实时分析免疫小鼠淋巴结中（绿色）高内皮小静脉和（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了解淋巴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有助于深入了解 LV 和 HEV 在免疫反应过程中如何调节、维持和协作，并深入了解炎症、移植、自身免疫、传染病和癌症中的淋巴管生成。公众健康相关性：对淋巴管生成和淋巴管功能的透彻了解将为淋巴水肿等情况下的淋巴功能和功能障碍提供治疗见解。淋巴管成像工具将深入了解免疫反应期间如何维持和调节该血管系统，并深入了解炎症、移植、自身免疫、传染病和癌症中的淋巴管生成。）