Analysis of uterine natural killer cell promotion of endometrial angiogenesis

子宫自然杀伤细胞促进子宫内膜血管生成的分析

• 批准号: 3219-2006
• 负责人: Croy, Anne
• 金额: $ 5.77万
• 依托单位: Queen's University
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2007
• 资助国家: 加拿大
• 起止时间: 2007-01-01 至 2008-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=361313
• 关键词: Analysis; uterine; natural; killer; cell

An unusual subset of immune cells (uNK cells) appears in the uterus in early pregnancy. In mice, these cells secrete signaling molecules (Interferon-gamma and Vascular Endothelial Cell Growth Factor) which bring about structural changes to maternal spiral arteries (SA), the blood vessels nourishing the placenta and fetus. We believe uNK cells guide blood vessel lining cells (endothelium) as new vessels form in implantation sites, directing them towards fetally-derived placental cells. Three Specific Aims will address our hypothesis. AIM 1 will characterize and correlate key molecular and functional properties of mouse SA during gestation days 8-12. Endothelial, vascular smooth muscle and position-defined uNK cells will be studied using laser capture microdissection, RNA isolation and Realtime quantitative RT-PCR. These molecular data will be correlated with functional data collected using our new technique for intravital microscopy of mouse SA. Uterine artery segments will be compared with control, structurally-stable intestinal vessels. AIM 2 will use microscopic and molecular approaches to define interactions between NK cells and migrating endothelial cells in vitro and in vivo. Lymphocyte-endothelial cell interactions and their regulation by other cell types and molecules will be studied in 3D hanging-drop co-cultures of human uterine microvascular endothelium and activated NK cells. Pregnancies in mice over-expressing interleukin 15 or with targeted gene deletions only in lymphocytes will be used to validate the in vitro results. AIM 3 will address when the uNK cell lineage diverges from NK cells in other tissues by comparing gene expression in decidual, splenic and marrow precursors. These studies model important aspects of human endometrial and fetal health and uterine angiogenesis in livestock where endometrial vascularity correlates with litter size. The data will advance our long-term goal of understanding the biology of uNK cells at the whole animal, cellular and molecular levels, and may provide insights into angiogenic roles of immune cells in general tissue healing or pathogenic remodeling.

妊娠早期子宫内出现一种不寻常的免疫细胞亚群（uNK细胞）。在小鼠体内，这些细胞分泌信号分子（干扰素- γ和血管内皮细胞生长因子），导致母体螺旋动脉（SA）的结构改变，而螺旋动脉是滋养胎盘和胎儿的血管。我们认为，uNK细胞引导血管内膜细胞（内皮细胞）在植入部位形成新的血管，引导它们向胎儿来源的胎盘细胞发展。三个具体目标将解决我们的假设。AIM 1将表征和关联妊娠8-12天小鼠SA的关键分子和功能特性。内皮细胞、血管平滑肌细胞和定位uNK细胞将使用激光捕获显微解剖、RNA分离和实时定量RT-PCR进行研究。这些分子数据将与使用我们的小鼠SA活体显微技术收集的功能数据相关联。将子宫动脉段与结构稳定的对照肠血管进行比较。AIM 2将使用显微镜和分子方法来定义NK细胞和迁移内皮细胞在体外和体内的相互作用。淋巴细胞与内皮细胞的相互作用及其受其他细胞类型和分子的调控将在人子宫微血管内皮和活化NK细胞的三维悬滴共培养中进行研究。过度表达白细胞介素15或仅在淋巴细胞中靶向基因缺失的小鼠妊娠将用于验证体外结果。AIM 3将通过比较个体、脾和骨髓前体的基因表达来解决NK细胞谱系何时与其他组织中的NK细胞分化。这些研究模拟了人类子宫内膜和胎儿健康以及家畜子宫血管生成的重要方面，其中子宫内膜血管分布与产仔数相关。这些数据将推进我们在全动物、细胞和分子水平上理解uNK细胞生物学的长期目标，并可能为免疫细胞在一般组织愈合或致病性重塑中的血管生成作用提供见解。